Protective effects of Suberoylanilide hydroxamic acid and Dapagliflozin administration on liver of diabetic rats

doi:10.21203/rs.3.rs-5015447/v1

Download PDF

Research Article

Protective effects of Suberoylanilide hydroxamic acid and Dapagliflozin administration on liver of diabetic rats

https://doi.org/10.21203/rs.3.rs-5015447/v1

This work is licensed under a CC BY 4.0 License

Version 1

posted

You are reading this latest preprint version

Background

Type 2 diabetes mellitus (T2DM) is common metabolic disorders. T2DM patients had 2-fold increase to get liver disorders. Evidence that some antidiabetic substances treated liver disorders in T2DM patients is evolving. Current study aimed to investigate hepatoprotective actions of Suberoylanilide hydroxamic acid (SAHA) and dapagliflozin (DAPA) in T2DM rats.

Methods

T2DM occured by high fat diet (HFD) and single Streptozotocin (STZ) injection (35 mg/kg i.p.). Forty rats sorted into 4 groups: NC (negative control), T2DM, T2DM + SAHA (5 mg/kg/i.p. for 8 weeks) and T2DM + DAPA (1mg/kg/p.o. for 8 weeks). At experimental end, levels of fasting blood glucose (FBG), fasting insulin, hepatic function tests [gamma glutamyl transferase (GGT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, albumin, total protein], lipid profiles [total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), triglyceride (TG)] measured in serum. Hepatic tissue homogenization prepared for estimating oxidative stress biomarkers [glutathione (GSH), malonaldehyde (MDA), superoxide dismutase (SOD)]. Hepatic histopathological examination made under light microscope.

Results

Diabetic rats had significant rise in liver weights and hepatic enzymes (AST, ALT, GGT, total bilirubin), lipid profile [TG, TC, LDL-C) in serum, and elevation in MDA in liver homogenate, but significant decline in total proteins, albumin, HDL-C) in serum and SOD, GSH in liver homogenate. These changes associated with histopathological changes in liver tissue as degeneration, vacuolation of hepatocytes, dilatation, and congestion of portal veins with lymphocytic infiltration. SAHA and DAPA treatment decreased liver weights, FBG, insulin, insulin resistance (IR), AST, ALT, GGT, bilirubin, TG, TC, LDL-C. SAHA and DAPA increased antioxidant enzymes (GSH, SOD) levels, serum total protein, albumin and HDL-C. Interestingly, DAPA was better that SAHA in improving liver enzymes, lipid profile, decreased FBG but SAHA was better in improving antioxidants as SOD, insulin levels and IR. Also, there were marked improvements in liver histopathological changes in SAHA and DAPA groups that were better in SAHA group.

Conclusions

Suberoylanilide hydroxamic acid and dapagliflozin represent an approach to protect liver versus DM-induced disorders via suppression oxidative stress, improve lipid profile, hyperglycemia and IR progression thus conserving liver functions and structure.

Dapagliflozin

liver

protective effects

rats

Suberoylanilide hydroxamic acid

type 2 diabetes mellitus

By 2045, there will likely be 700 million more diabetes mellitus (DM) patients worldwide than 463 million currently cases [1]. More than 90% of all people with diabetes are diagnosed with type 2 diabetes mellitus (T2DM), a complicated metabolic condition linked to insulin resistance (IR) and pancreatic dysfunction [2]. Chronic hyperglycemia with various causes and disorders in metabolism of fats, proteins, and carbohydrates are T2DM hallmarks. There is a strong correlation between chronic hyperglycemia and a high rate of complications, including liver disorders as cirrhosis, hepatocellular cancer, nonalcoholic fatty liver disease (NAFLD), acute liver cell failure and abnormal liver enzymes [3]. A number of microvascular and macrovascular problems may result from improper T2DM therapy and control [4]. Even with the abundance of antidiabetic drugs available, T2DM is still linked to a high rate of morbidity and death.

Histone deacetylases (HDAC) and histone acetyltransferases (HAT) have drawn a lot of interest as crucial molecules involved in controlling a range of cellular responses and altering pathological situations [5]. Transcriptional activation and nucleosome relaxation follow chromatin structural alteration brought about by histone acetylation mediated by histone acetylation transferase (HAT). It follows that a crucial mechanism for epigenetic gene expression regulation is provided via nucleosome histone tails acetylation. Transcriptional repression, chromatin condensation, and de-acetylation are brought about by the opposite process that HDACs mediate [6]. Histone deacetylase enzymes have been the subject of several investigations throughout the last ten years because they have a pathogenic role in DM [6]. HDACs can epigenetically regulate carbohydrates and fatty acids metabolism by regulating many genes transcription. Classical HDACs could affect non-histone protein deacetylation, signal transducer, transcription 3 (STAT3) activation that leads to inhibition of gluconeogenesis enzymes expression. There is evidence linking diabetes mellitus (DM) to HDAC inhibitors (HDACi), which have been demonstrated to have positive actions on β-cell formation, differentiation, proliferation, and functions in addition to signaling of insulin [7]. Furthermore, in vivo and in vitro models of cytokine-induced β-cell destruction are prevented by HDACi [8, 9]. Suberoylanilide hydroxamic acid (SAHA), commonly referred to as Vorinostat or Vorino, is an HDAC inhibitor with anti-inflammatory and antioxidant activities that primarily suppress Class II, I, and IV HDACs. It approved in 2006 for cutaneous T-cell lymphoma treatment [10]. SAHA had antidiabetic benefits in type 1 diabetes mellitus (T1DM) animal model [11]. SAHA prevents liver fibrosis in LX2 cells by blocking HDAC2, HDAC6, and HDAC8 proteins [6].

Sodium–glucose cotransporter-2 inhibitors (SGLT2is) were promising pharmaceuticals class for T2DM therapy because they improve insulin sensitivity, suppress gluconeogenesis, and reduce glucose reabsorption by blocking SGLT2 protein that included in 90% of glucose reabsorption [12]. The FDA originally authorized canagliflozin in 2013 and dapagliflozin (DAPA) in 2014 as SGLT2i [13]. In addition to their anti-hyperglycemic action, SGLT2i has been associated in several trials to improve hepatic function in individuals with T2DM [14]. Preclinical studies showed that SGLT2i decrease liver lipid accumulations [15] and decrease fat mass and body weights by enhancing fatty acids utilization instead of glucose as source of energy for metabolism [16]. Due to the insulin-independent mechanism of SGLT2i, individuals with insulin resistance or reduced pancreatic function may benefit from this family of drugs as an additional treatment. So, SGLT2is may be appropriate in treatment of T2DM [17].

This experimental research was conducted to study possible protective effects of histone deacetylase inhibitors (Suberoylanilide hydroxamic acid) and Sodium–glucose cotransporter-2 inhibitor (dapagliflozin) in vivo administration on liver of T2DM rat’s model and examine the underlying mechanism of their effects.

Animals

Forty adult male Albino rats weighing 250–300 gm and aged 8 weeks obtained from Animal house at Faculty of Pharmacy, King Abdulaziz University (KAU), Jeddah, Saudi Arabia. Rats kept in suitable laboratory status of light (12h light/dark cycle), temperature (25 ± 1°C), and humidity 60% with ad libitum access to distilled water and a rodent rat’s diet for one week before experiment start for acclimatization to laboratory condition. The experimental procedure made according to ARRIVE (Animal Research Reporting of In Vivo Experiments) and approved by Ethical Committee of King Abdulaziz University, Jeddah, Saudi Arabia.

Chemicals and drugs

Streptozotocin (STZ) bought from Sigma‑Aldrich (Sigma-Aldrich Co. St. Louis, Missouri, USA). STZ mixed in 0.1M citrate buffer (pH 4.4) and administered to rats intraperitoneally (i.p.). Suberoylanilide hydroxamic acid bought from Selleck Chemicals (Sigma-Aldrich Co. St. Louis, Missouri, USA), and mixed in vehicle of 10% dimethyl sulfoxide (DMSO), 45% polyethylene glycol 400 (PEG-400), 45% saline & administered i.p. Dapagliflozin bought from AstraZeneca Pharmaceutical Company (Cairo, Egypt), and put in 0.5% aqueous solution of carboxymethyl cellulose (CMC) and ingested oral via gastric lavage.

Experimental design and induction of T2DM

Forty rats randomly sorted equally into 4 groups as follows: first group was negative control (NC) that fed by regular chow diet (23% protein, 53% carbohydrate, and 5% fat) for 12 weeks. From week 4, NC group rats took 1 ml/kg daily i.p. injections of 10% DMSO, 45% PEG-400, 45% saline (vehicle of SAHA) and 1 ml/kg daily orally of CMC (vehicle of DAPA). All other rats (n = 30) were fed high fat diet (HFD) (normal diet + 2.5% cholesterol + 20% saccharose + 15% lard stearin) for 11 weeks and received at 3rd week a once STZ injection (35 mg/kg i.p.) for T2DM induction [18]. Three days followed STZ intake, fasting blood glucose (FBG) levels evaluated and animals showed FBG ≥ 250 mg/dl considered diabetic and included into research [19]. For more 8 weeks, HFD gave with the following therapy. Second group was T2DM and took saline [20]. Third group (T2DM + SAHA) received daily SAHA injection (5 mg/kg /i.p. for 8 weeks) [21]. Fourth group (T2DM + DAPA) received daily dapagliflozin (1mg/kg/p.o. for 8 weeks) [22].

Body weights of rats estimated at beginning of experiment and weekly using digital scale. Weight gain (%) calculated as final body weight – initial body weight divided by initial body weight then multiplied by 100. Food intake was calculated weekly and food efficiency ratio (FER) (%) calculated as weight gain (g/week) divided by intake of food (g/week) X 100.

At experimental end, rats were fasted for 12 hours. Thiopental sodium (50 mg/kg) was used to anesthetize rats. Retro-orbital plexus, the orbital sinus, was the site of blood sample collection. Blood collected into plain tubes then left to coagulate for 20 minutes. Sera were obtained by centrifuging the blood for 15 minutes at 4000 rpm. The separated sera were stored frozen at -20°C in aliquots till used.

Rats were died by cervical dislocation and livers isolated, washed with cold saline and weighed. Hepatic index (%) calculated as weight of liver divided by final body weight X 100. Liver was dissected into two halves. One liver half fixed in 10% formalin for histopathological evaluation under light microscopy. Other liver half washed with saline, frozen in liquid nitrogen, and kept at – 80°C till preparation of hepatic homogenates.

Hepatic tissue homogenate preparation

Half of liver homogenized in phosphate-buffered saline (PBS) to made 10% (w/v) homogenate utilized homogenizer (Omni International, Kennesaw, GA, USA). Homogenates further spun at 5,000g at 4°C for 15 min. Supernatants collected, aliquoted and storage at -20° for further measurement of oxidative stress biomarkers.

Biochemical Measurements

FBG was measured using colorimetric kit (catalog# ab282922) from Abcam (United Kingdom). Fasting serum insulin concentrations estimated by rat insulin enzyme-linked immunosorbent assay (ELISA) kit (catalog# MBS281388) from My BioSource (San Diego, USA) based upon manufacturer’s protocol. Assessment of IR carried out using homeostatic model assessment of insulin resistance (HOMA-IR) index. HOMA-IR = [fasting insulin (µU/ml) × fasting plasma glucose (mg/dl)]/405 [23]. Liver functions as alanine aminotransferase (ALT), total bilirubin, gamma glutamyl transferase (GGT), aspartate aminotransferase (AST), total protein and albumin estimated by colorimetric method by available kits (Teco diagnostics, Cairo, Egypt). Serum levels of lipid profile as total cholesterol (TC), triglycerides (TG), high-density lipoprotein concentration (HDL-C) estimated by kits (Spectrum diagnostics, Cairo, Egypt) based upon manufacturer’s protocol. Low density lipoprotein concentration (LDL-C) calculated according to Friedewald formula: LDL-C = TC – [HDL-C + TG/5)] [24]. Values of malondialdehyde (MDA) (catalog# MBS268427), lipid peroxidation marker, and superoxide dismutase (SOD) (catalog# MBS036924) and glutathione (GSH) (catalog# MBS265966), markers of antioxidants estimated in hepatic homogenates by commercially available kits from MyBioSource, Inc. (San Diego, CA 92195 − 3308, USA) based upon manufacturer’s protocols.

Histological examination

The livers were examined closely for any noticeable alterations. Portions of left liver lobes prepped for light microscopy so that a liver histological assessment may be carried out. After being first preserved in a 10% neutral formalin solution, sample progressively dried out in 50% – 100% ethanol, cleaned in xylene, and then embedded into paraffin. Hematoxylin and eosin (HX&E) staining made after sections cut into 5 m thick portions. Slices of liver were analyzed by a skilled histopathologist using light microscopy to assess tissue alterations.

Statistical Analysis

Results presented as mean ± standard deviation (SD). Value analysis made using IBM Corporation's SPSS version 22 (Armonk, NY, USA). Shapiro-Wilk test employed to determine the normality of value distributions. Tukey's test is utilized to compare groups of normally distributed values after One-Way ANOVA has been used to analyze the data. Kruskal Wallis and Mann Whitney tests utilized to compare groups when values are abnormally distributed (weight increase, weight index, FER, liver weights, and liver indices). Statistical significance defined as a p < 0.050.

Biological results

Compared to NC group, there were significant increase in T2DM, T2DM + SAHA and T2DM + DAPA groups of final body weight (p < 0.001, p < 0.001 and p < 0.010, respectively), Weight gain (p < 0.001, p < 0.010 and p < 0.050, respectively) and weight gain percentage (p < 0.001, p < 0.050 and p < 0.050, respectively). Food Efficiency Ratio (FER) significantly decreased in T2DM, T2DM + SAHA and T2DM + DAPA groups versus NC group (p < 0.001, p < 0.010 and p < 0.050, respectively). The liver weight significantly decreased in T2DM + DAPA versus negative control (p < 0.010). The hepatic index significantly increased in T2DM compared with NC and T2DM + DAPA groups (p < 0.001 and p < 0.010) (Table 1).

Table 1

Comparison of initial, final body weight, weight gain, Food Efficiency Ratio, liver weight and liver index in various studied groups.
Variables	NC group (n = 10)	T2DM group (n = 10)	T2DM + SAHA group (n = 10)	T2DM + DAPA group (n = 10)
Initial body weight (grams)	272.70 ± 45.97	232.80 ± 35.37	235.90 ± 29.12	252.10 ± 14.04
Final body weight (grams)	404.50 ± 50.31	248.40 ± 41.30^***	287.40 ± 56.90^***	308.20 ± 72.21^**
Weight gain (grams)	131.80 ± 55.23	15.60 ± 36.44^***	51.50 ± 64.40^**	56.10 ± 67.21^*
Weight gain (%)	32.07 ± 11.69	5.32 ± 12.74^***	15.35 ± 18.79 ^*	14.84 ± 17.05^*
Food Efficiency Ratio (FER) (%)	0.051 ± 0.022	0.006 ± 0.014^***	0.020 ± 0.024^**	0.021 ± 0.026^*
Liver weight (grams)	9.76 ± 1.16	8.75 ± 1.09	8.67 ± 1.33	6.98 ± 2.18 ^**
Liver index (%)	2.42 ± 0.16	3.62 ± 0.82^***	3.17 ± 0.90	2.35 ± 0.77^##
Data expressed as mean +/- standard deviation. ^: significance versus NC group; ^#: significance versus T2DM. : P < 0.050, : P < 0.010, *: P < 0.001.

Food intake at different weeks among different studied groups is shown in Fig. 1.

Glycemic control results

At the 3rd week, FBG level was significantly increased in T2DM, T2DM + SAHA and T2DM + DAPA groups versus negative control (p < 0.0001 for all). At the 11th week, blood glucose level significantly declined in T2DM + DAPA group versus T2DM and T2DM + SAHA groups (p < 0.001 for both) and in T2DM + SAHA versus T2DM (p = 0.029). Fasting insulin levels and HOMA-IR significantly elevated in T2DM, T2DM + SAHA and T2DM + DAPA groups versus negative control (p < 0.0001 for all) but were significantly decreased in T2DM + SAHA and T2DM + DAPA groups versus T2DM group (p < 0.0001 for all) and in T2DM + DAPA groups versus T2DM + SAHA (p < 0.0001 for both) (Table 2).

Table 2

Comparison of glycemic control in various studied groups.
Variables	NC group (n = 10)	T2DM group (n = 10)	T2DM + SAHA group (n = 10)	T2DM + DAPA group (n = 10)
Blood glucose (mg/dl)
3rd week	75.70 ± 7.60	395.20 ± 31.95^***	403.70 ± 48.87^***	410.90 ± 57.36^***
11th week	78.90 ± 9.98	354.90 ± 22.57^***	333.20 ± 7.50^***,#	263.70 ± 20.82^***,###,&&&
Insulin (uIU/ml)	26.80 ± 2.30	85.40 ± 3.06^***	42.50 ± 1.84^***,###	75.30 ± 2.11^***,###,&&&
HOMA-IR	5.22 ± 0.76	74.94 ± 6.90^***	34.97 ± 1.85^***,###	49.01 ± 3.80^***,###,&&&
Data expressed as mean +/- standard deviation. ^: significance versus NC group; ^#: significance versus T2DM, ^&: significance versus T2DM + SAHA group. : P < 0.050, ***: P < 0.001.

Liver function test results

At experimental end, AST, ALT and GGT values were significantly increased in T2DM, T2DM + DAPA and T2DM + SAHA groups versus NC (p < 0.001 for all) and in T2DM versus T2DM + DAPA and T2DM + SAHA groups (p < 0.001 for all). Meanwhile, AST and ALT values were significantly decreased in T2DM + DAPA than T2DM + SAHA group (p < 0.001). Total bilirubin was significantly elevated in T2DM versus negative control, T2DM + SAHA and T2DM + DAPA (p < 0.001 for all) and in T2DM + DAPA versus negative control (p < 0.050). Total proteins levels declined in T2DM, T2DM + SAHA and T2DM + DAPA compared to NC (p < 0.001 for all); decreased in T2DM versus T2DM + SAHA and T2DM + DAPA (p < 0.001), declined in T2DM + DAPA versus T2DM + SAHA (p < 0.001). Albumin levels significantly decreased in T2DM and T2DM + DAPA versus NC (p < 0.001 for all); decreased in T2DM versus T2DM + SAHA and T2DM + DAPA (p < 0.001), declined in T2DM + DAPA than T2DM + SAHA (p < 0.001) (Table 3).

Table 3

Comparison of liver function tests and lipid profile in various studied groups.
Parameters	NC group (n = 10)	T2DM group (n = 10)	T2DM + SAHA group (n = 10)	T2DM + DAPA group (n = 10)
AST (U/L)	116.10 ± 4.06	248.80 ± 4.32^***	171.60 ± 8.58^***,###	151.50 ± 6.77^***,###,&&&
ALT (U/L)	80.70 ± 2.31	150.60 ± 4.27^***	127.50 ± 5.08^***,###	115.40 ± 4.86^***,###,&&&
GGT (U/L)	5.98 ± 1.58	23.30 ± 2.41^***	13.00 ± 1.83^***,###	13.90 ± 1.29^***,###
Total bilirubin (µmol/L)	5.58 ± 0.56	8.36 ± 0.80^***	5.94 ± 0.39^###	6.38 ± 0.41^*,###
Total protein (mg/dl)	72.16 ± 2.36	32.30 ± 2.50^***	48.60 ± 2.91^***,###	43.90 ± 2.42^***,###,&&&
Albumin (mg/dl)	45.20 ± 2.65	23.50 ± 1.78^***	42.50 ± 3.44^###	33.90 ± 2.42^***,###,&&&
Triglyceride (mg/dl)	132.90 ± 2.96	375.00 ± 9.13^***	314.00 ± 12.65^***,###	265.00 ± 15.81^***,###,&&&
Total cholesterol (mg/dl)	93.70 ± 3.06	258.70 ± 7.67^***	191.00 ± 13.29^***,###	136.50 ± 9.73^***,###,&&&
LDL-C (mg/dl)	44.70 ± 3.20	151.10 ± 7.81^***	74.90 ± 7.17^***,###	77.80 ± 4.21^***,###
HDL-C (mg/dl)	72.30 ± 2.71	32.60 ± 3.95^***	36.10 ± 4.12^***	36.00 ± 4.62^***
Data expressed as mean +/- standard deviation. ^: significance versus NC group; ^#: significance versus T2DM, ^&: significance versus T2DM + SAHA group. : P < 0.050, ***: P < 0.001.

Lipid profile results

At experimental end, Triglyceride, total cholesterol, LDL-C values significantly elevated in T2DM, T2DM + DAPA and T2DM + SAHA groups versus NC (p < 0.001) and in T2DM versus T2DM + DAPA and T2DM + SAHA groups (p < 0.001 for all). Triglyceride and total cholesterol serum values were significantly increased in T2DM + SAHA versus T2DM + DAPA (p < 0.001). HDL-C values were significantly declined in T2DM, T2DM + DAPA and T2DM + SAHA groups versus NC (p < 0.001 for all) (Table 3).

Oxidative stress results

At experimental end, MDA liver homogenate levels were significantly elevated in T2DM, T2DM + DAPA and T2DM + SAHA groups versus NC (p < 0.001 for all) and in T2DM versus T2DM + DAPA and T2DM + SAHA groups (p < 0.001 for both). SOD and GSH liver homogenate values were significantly declined in T2DM, T2DM + DAPA and T2DM + SAHA groups versus NC (p < 0.001 for all); decreased in T2DM versus T2DM + DAPA and T2DM + SAHA groups (p < 0.001 and p < 0.050). Meanwhile, SOD value was significantly increased in T2DM + SAHA versus T2DM + DAPA (p < 0.001) (Table 4).

Table 4

Comparison of oxidative stress markers in various studied groups.
Parameters	NC group (n = 10)	T2DM group (n = 10)	T2DM + SAHA group (n = 10)	T2DM + DAPA group (n = 10)
MDA (nmol/ml)	9.67 ± 3.33	23.54 ± 2.38^***	15.31 ± 2.14^***,###	17.55 ± 1.82^***,###
SOD (U/ml)	17.93 ± 2.11	9.11 ± 1.37^***	14.40 ± 0.71^***,###	10.93 ± 1.15^***,#,&&&
GSH (U/L)	315.99 ± 13.04	225.01 ± 11.60^***	267.27 ± 7.25^***,###	277.01 ± 10.72^***,###
Data expressed as mean +/- standard deviation. ^: significance versus NC group; ^#: significance versus T2DM, ^&: significance versus T2DM + SAHA group. : P < 0.050, ***: P < 0.001.

Histology results

The livers of healthy control rats were examined using HX and E staining, and the results showed a typical hepatic shape with the liver split into classic hepatic lobules. Hepatocyte cords that extend outward from central vein to lobule's perimeter make up the classic lobule. Loose stromal connective tissue envelops portal triads, also known as tracts, which are dispersed at the corners of lobules. The polyhedral hepatocytes had one or two conspicuous nucleoli, a clear vesicular spherical nucleus, and a highly eosinophilic cytoplasm speckled with basophilic granules. Binucleation of hepatocytes occurs often. Flat endothelial cells line central vein, and statistically significant hepatic blood sinusoids found between endothelial cell-lined hepatic cell cords and von Kupffer cells with ovoid nuclei (Fig. 2a). Peripherally each lobule has three to six portal areas with more fibrous connective tissue, each of which has three interlobular structures that comprise portal triad or portal tracts, portal vein, hepatic artery and one or two small branches of bile ducts lined with cuboidal epithelium (Fig. 2.b).

Rats of diabetic group showed severe damage in rat liver. The most significant alterations were disorderly hepatocyte, inflammatory, degenerative, necrotic, nucleus karyolysis and hyperplastic changes. Liver showed degenerations and vacuolation of hepatocytes and some of them exhibited one large cytoplasmic vacuole. Severe infiltrative fatty changes in form of more well-defined fat droplets occupying hepatocytes cytoplasm, pushing nucleus to periphery (Fig. 3a). Also, multiple inflammatory cells appeared. Hydropic degeneration of hepatocytes was observed around dilated central vein. Central vein showed dilatation and congestion with leukocytic infiltrations around it (Fig. 3b). There were many localized necrotic regions with extensively vacuolated hepatocyte cytoplasm and strongly stained pyknotic nuclei, together with a noticeable infiltration of mononuclear cells or widespread hepatic coagulative necrosis (Figs. 3a,b). Portal tract showed congested and dilated portal vein and mononuclear leucocytic cell infiltrations around portal vein (Fig. 3c).

Treated diabetic groups with SAHA showed improvement in hepatic histological structure. Amelioration of hepatocytes, and alleviate inflammation, leucocytic cell infiltration, necrotizing hepatocytes, which is produced by STZ. Liver tissue kept its normal hepatic shape with radiating hepatic cell cords and central veins to some extent similar to negative control group (Fig. 4a). Most of hepatocytes appeared polyhedral with strongly eosinophilic cytoplasm dotted with basophilic granules, and distinct vesicular rounded nuclei and one or two prominent nucleoli. Central vein is lined with flat endothelial cells and hepatic blood sinusoids were noticed between hepatic cell cords lined with endothelial cells and von Kupffer cells. Portal tract showed normal bile duct, hepatic artery and portal vein (Fig. 4b).

Regarding diabetic rat treated with DAPA showed moderate improvement in liver histological structure. Liver tissue preserved to some extent its normal hepatic shape with central veins and radiating hepatic cell cords to some extent as negative control group. Most of hepatocytes appeared polyhedral with eosinophilic cytoplasm dotted with basophilic granules, and vesicular rounded nuclei. Central vein is lined with flat endothelial cells and hepatic blood sinusoids noticed between hepatic cell cords lined with endothelial cells. Portal tracts showed normal histological structure as that of control group (Figs. 5a,b). Some specimens still showed moderate fatty infiltrative changes where smaller well-defined fat droplets occupying hepatic cytoplasm with loss of normal architecture of liver tissue and mononuclear cell infiltration's (Figs. 5c,d).

T2DM-related metabolic abnormalities may cause liver damage, which may ultimately lead to a number of liver illnesses, including cirrhosis, hepatocellular carcinoma, and fatty liver [25]. Numerous variables, including hyperglycemia, insulin resistance, dyslipidemia, oxidative stress, and inflammations, can lead to diabetic liver damage [25].

In this study, HFD/STZ utilized to induce a T2DM liver damage rat model [26]. The results of this research revealed that, final body weight, weight gain, weight gain percentage and FER declined in T2DM group compared to negative control group. In this respect, According to Zhu et al., T2DM rat group's body weight dramatically dropped while their intake of food, water, and urine volume were significantly elevated [26]. Lower body weight in T2DM is due to catabolic state of poorly controlled glycaemia. Under these conditions, lipolysis, metabolic processes, and oxidative degradation of amino acids elevated; they degrade greatest energy and tissue reserves in animal, so decreasing body weights [27]. In this study, body weights in diabetic rats were not improved by administration of SAHA or DAPA to diabetic rats, which may be caused by continuous catabolic condition. SAHA appears to have complex effects on body weight. While some metabolic alterations and adverse consequences, such as anorexia, were noted in preclinical and clinical trials, significant weight changes were not always recorded [28, 29]. This suggests that not all circumstances and dose ranges will result in a consistent or direct effect of SAHA on body weight. In the meantime, osmotic drainage brought on by the glycosuria brought on by DAPA alters body composition, resulting in a decrease in body fat and body weight loss. According to Phrueksotsai et al., DAPA prescription resulted in significant decline in body weight and body fats after 12 weeks of therapy, with a mean 3% decrease in baseline body weights. Furthermore, correlation analysis showed a significant positive relationship between weight loss and decrease in liver fat content [30]. Ahmed et al. found that total body weight was declined in three groups of rats that received three different DAPA doses (0.75, 1.5 and 3 mg/kg, p.o.) than negative and diabetic control groups [31].

Liver index in this research was significantly increased in T2DM versus negative control and T2DM + DAPA groups. Zhu et al. reported that the liver indices were significantly increased in T2DM rat’s model [26]. In this study, the liver weight significantly decline in T2DM + DAPA versus negative control. DAPA medication significantly reduces total liver fat content in T2DM patients [32]. In addition to decreased body weight, metabolic substrate shift from glucose to fatty acids and likely higher hepatic oxidation of fatty acids are other theories explaining the loss of liver fat [33].

This study showed that at experimental end FBG level, fasting insulin level and HOMA-IR significantly elevated in T2DM group versus negative control as previously reported [26, 34]. In rats, HFD causes IR [26]. The results of this research showed that FBG, fasting insulin, HOMA-IR was significantly decreased T2DM + SAHA versus T2DM but were still significantly elevated than negative control group. Bocchi et al. reported that injection of SAHA (25 mg/kg i.p.) for 23 days into mice leads to improved sensitivity to insulin [35]. Silva et al. reported that injection of SAHA (25–50 mg/kg i.p.) for 8 weeks into HFD fed mice with leads to ameliorate insulin sensitivity [36]. In the mesenchymal stem cell model MG63, SAHA was shown to affect the formation of β-cells and improve insulin production by upregulating transcription factor PDX1 expression. Additionally, pretreatment with SAHA increased β-cell markers in response to high glucose challenges, indicating that it may improve stem cells' ability to develop into β-cells that produce insulin [37]. Histones, other regulatory proteins and different transcription factors that either indirectly or directly included in metabolism of glucose are deacetylated by HDACs. Furthermore, histone acetylation regulates the glucose-mediated regulation of insulin gene transcription, indicating HDACs involvement in manufacture and function of insulin [38]. The results of this research revealed that therapy of diabetic rats with DAPA led to decline in FBG, fasting insulin, HOMA-IR versus T2DM but were still significantly higher than negative control group. In DAPA treated diabetic rats, FBG was significantly decline, but fasting insulin and HOMA-IR were increased versus SAHA treated diabetic group. Tang et al. reported that after four weeks of therapy, there was decline in blood glucose values and an increase in urine glucose excretion with DAPA [39]. Joannides et al. reported that DAPA administration for 45 days into PEPCK transgenic rats caused reduced plasma glucose and insulin values due to improve IR, elevated fat and muscle glucose uptake and GLUT4 protein values, and decreased size of adipocyte and elevated number of adipocyte, but not elevated secretion of insulin [40]. HFD/STZ -induced diabetic mice treated with DAPA showed improved glucose tolerance, IR and insulin secretion, with a significant elevation in insulin content of pancrease [41].

Liver was affected in T2DM rat models in this study as revealed by increased ALT, AST, GGT, total bilirubin, and decline in total protein and albumin versus negative control group. Zhu et al. reported ALT and AST values were significantly elevated in T2DM rats model [26]. SAHA administration to diabetic rats had protective effect on liver as revealed in this study by decreased AST, ALT, GGT, total bilirubin and significant increase in total protein and albumin. SAHA has protective effect on liver damage produced by lethal hemorrhagic shock in rats that was associated with elevated H3K9 acetylation and suppression of JNK/caspase-3 apoptotic pathway [42]. SAHA administration significantly reduced serum values of AST, ALT, and lactate dehydrogenate, increased the survival rate and decreased apoptotic markers expression in liver tissues, suggesting its protective role in early hemorrhagic shock conditions [43]. Wang et al. found that in rats carbon tetrachloride (CCl4) significantly made liver fibrosis and elevated serum values of transforming growth factor (TGF)-β1, total bilirubin, ALT, AST, laminin, and procollagen type III; liver HDAC2, p-Smad2/3, HDAC6, HDAC8, α-SMA and connective tissue growth factor (CTGF) proteins; whereas Smad7 mRNA and AH3 protein levels were notably suppressed. SAHA treatment significantly downregulated, these liver chemistries, cytokines and liver fibrosis-related genes and mitigated hepatic fibrosis [44]. Alhaddad et al. reported that SAHA administration (15 mg/kg/day p.o.) for 8 weeks to rat model of autoimmune hepatitis made by Concanavalin A led to decreased in AST and ALT liver enzymes [45]. DAPA administration to diabetic rats had protective effect on liver as revealed in this study by decreased ALT, AST, GGT, total bilirubin and significant increase in albumin and total protein. Dapagliflozin therapy protected the liver in db/db mice, as revealed by markedly lower levels of oxidative stress and inflammatory indicators, hepatic lipid buildup, and plasma ALT activity and TG levels [39]. DAPA markedly reduced ALT and AST serum levels and protected liver from pathologic damages in diabetic mice [46]. A clinical trial utilizing DAPA in T2DM patients revealed reduced values of liver damage biomarkers, as ALT, AST, and GGT; combining carboxylic acids (OM-3CA) resulted in a significant decline in hepatic fat content [32]. DAPA led to ALT-lowering effect by decrease liver fat deposition by making hyperglucagonemia [47] and ameliorate IR caused by decreased ectopic steatosis [48].

Dyslipidemia is considered a major cardiovascular disorders risk factor resulting in myocardial infarction, sudden cardiac arrest, and death. In this study, significant increase in serum TC, TG and LDL-C values accompanied by decline in serum HDL-C value were demonstrated in T2DM rats versus control group as reported by others [26, 34]. IR and dyslipidemia are important risk factors of diabetic hepatic damage [25]. In this study, treatment of diabetic rats with SAHA led to a significant decreased in serum TG, TC and LDL-C values compared to T2DM rats but were still significantly elevated versus control group. SAHA intake did not improve levels of HDL-C. SAHA exhibits significant anti-inflammatory effects by reducing pro-inflammatory cytokines production as interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and interferon gamma. These cytokines are known to influence metabolic processes, including lipid metabolism [49]. HDACs inhibition by SAHA affects various signaling pathways and gene expressions related to lipid biosynthesis and metabolism as lipoxygenases [50]. The induction of 15-lipoxygenase-1 by SAHA, for example, correlates with significant changes in lipid metabolism [51]. However, direct studies specifically focusing on serum lipid profiles post-SAHA treatment are limited, indicating a need for more targeted research in this area. In this study, treatment of diabetic rats with DAPA led to a significant decreased in serum TG, TC and LDL-C values versus T2DM rats but were still significantly elevated than control group. Decline in TG and TC levels were more effective that SAHA administration. Meanwhile, DAPA administration did not improve HDL-C levels. Leng et al. found that DAPA decreased the values of TG and free fatty acids (FFAs) in liver and serum, which correlated with decreased lipotoxicity in diabetic mice [46]. Hazem et al. reported lower liver weights and decreased serum values of TG and TC in rats received DAPA [52]. Ahmed et al. reported that TC and TG were decreased in three groups that received three different DAPA doses (0.75, 1.5 and 3 mg/kg, p.o.) for 6 weeks more than negative and diabetic control groups [31]. This reduction in TC and TG may be attributed to the total reduction in body weight or may be explained by shift of metabolic substrate from glucose to fatty acids [53].

The development of T2DM and its consequences, as well as IR, are significantly affected by oxidative stress. Reactive oxygen species (ROS) produced by dyslipidemia and hyperglycemia may damage live cells and certain cell membrane receptors, which may lead to damage to organs including liver [54]. The results of this research showed elevation of MDA and decrease in SOD and GSH in diabetic rats versus negative control group. Researches revealed that tissue homogenate GSH concentrations in STZ-induced diabetic rats significantly decline versus negative control rats [55, 56]. Decline GSH and SOD values in diabetic rats caused by its increased consumption that is required to relieve oxidative stress. Zhu et al. revealed that oxidative stress and inflammation significantly elevated in hepatic homogenates of T2DM rats [26]. Hazem et al. revealed that diabetic group had much lower values of GSH, SOD, and catalase and significantly greater amounts of MDA [52]. By further reducing tissue oxidative stress, tight glycemic control or add-on techniques which block oxidative stress to antihyperglycemic medications might improve capacity to prevent disease development in organ damage due to diabetes [39]. In this study, diabetic rats that received SAHA showed significant decrease in MDA but elevation of SOD and GSH hepatic homogenate levels versus T2DM rats. Bakhdar et al. reported that SAHA administration to Wistar rats (15 mg/kg/day i.p.) for 28 days led to pancreatic protection via anti-inflammatory and antioxidant actions [57]. SAHA was shown to reduce hepatic cellular injury and ROS production in lipopolysaccharide (LPS)-induced liver damage. It enhanced the antioxidant enzyme GSH and inhibited apoptotic signaling pathways, suggesting its potential in alleviating inflammatory liver conditions [43]. In this study, diabetic rats that received DAPA showed significant decrease in MDA but elevation of SOD and GSH hepatic homogenate levels versus T2DM rats. The effect of SAHA in elevation of SOD hepatic homogenate level was better than DAPA. Hazem et al. provides evidence for hepatoprotective effects of DAPA, as seen by the dose-dependent increases in antioxidant enzymes SOD, catalase activity, and GSH and decline in MDA levels [52]. In Kashiwagi study, the SOD activity was significantly improved after 8 weeks of diabetic group treatment with insulin in addition to dapagliflozin [58]. The results of this research suggest that dapagliflozin significantly improved antioxidant status in diabetic rats. The treatment's antihyperglycemic impact, which lessens the load of oxidative stress, might be the cause of this improvement.

In this study, diabetic rats showed severe liver damage. The most significant alterations in diabetic control rat liver were disorderly hepatocyte, inflammatory, degenerative, necrotic, nucleus karyolysis and hyperplastic changes. These findings are in accord with those of Ahmed et al. who found that in diabetic, liver severe infiltrative fatty changes in form of more well-defined fat droplets occupying cytoplasm of hepatocytes, pushing nucleus to periphery. Also, multiple inflammatory cells appear with loss of normal architecture of hepatocytes [31]. Also, Salih et al. [59] observed that in STZ-diabetic mice showed more progressive changes, sever congestion, necrotic foci, hydropic changes, fatty changes in hepatocytes and aggregation of lymphocytes between the hepatocytes. Furthermore, additional research has documented damage to the liver cells, sinusoidal dilatation in the central venous region, elevated cell apoptosis, and elevated lipid droplets in the liver cells of diabetic mice [60]. By activating NF-B that stimulates pro-apoptotic genes activity in liver cells and increases the creation of ROS, hyperglycemia circumstances will aggravate the process of liver damage by inducing oxidative stress and inflammatory conditions [61].

Our results showed that treated diabetic groups with SAHA showed improvement in liver histological structure. Amelioration of hepatocytes, and alleviate inflammation, leucocytic cell infiltration, necrotizing hepatocytes, which is produced by STZ. So, hepatic tissue maintained its normal hepatic lobular shape with central veins and radiating hepatic cell cords to some extent similar to negative control group. Zhao et al. [62] evaluate SAHA-mediated protection against LPS-induced hepatic damage using histological examination. As predicted, the 24-hour LPS exposure significantly increased the inflammatory response in the murine liver. This was demonstrated by the inflammatory cells' growing penetration into parenchyma, where necrotic and apoptotic hepatocyte cell death were noted. Compared to LPS alone group, liver architecture was better conserved in the mice treated with SAHA following an LPS injection. SAHA prevented ROS from being produced by LPS and stopped the antioxidant enzyme glutathione from declining due to STZ. Additionally, SAHA reduced the hepatic apoptosis caused by STZ. Additionally, SAHA prevented the activation of mitogen-activated protein kinases p38 and Jun N-terminal kinase, as well as redox-sensitive kinase and apoptosis signal-regulating kinase-1 [62]. Our findings demonstrate that SAHA can mitigate the hepatotoxicity caused by STZ and imply that a novel therapeutic approach for treating STZ-induced inflammatory conditions could involve blocking the upstream processes necessary for the function of apoptotic signal-regulating kinase-1. Also, Bocchi, et al. [63] discovered that by lowering cell oxidative damage, SAHA therapy might reverse the initial functional aberration in cardiomyocytes. After being exposed to SAHA, diabetic cardiomyocytes (CMs) showed a simultaneous reduction in metabolic status, namely NAD(P)H dehydrogenase activities and ROS values. This finding raises the possibility of a mechanistic relationship between changed ROS generation and cell metabolism. In diabetic cells, SAHA aids in the restoration of proper redox signaling, which is necessary for maintaining homeostasis of cardiomyocyte and is implicated in heart's action to stress [64].

The present research showed that diabetic rat reveived DAPA showed in some specimens, moderate improvement in liver histological structure. On the other hand, some specimens still showed moderate fatty infiltrative changes with droplets of fats occupying hepatocytes cytoplasm and loss of normal architecture of liver tissue accompanying mononuclear cell infiltrations. Our data agree with the results of Ahmed et al. [31] who noticed that diabetics on DAPA (0.75 mg/kg), section of rat liver tissue showed mild to moderate fatty infiltrative changes where smaller well-defined fat droplets occupying hepatic cytoplasm with loss of liver tissue normal architecture. Diabetic rats on DAPA 1.5 and 3 mg/kg groups: section of rat hepatic tissue revealed improvement of fatty infiltrative alteration with normal hepatocytes around central vein and normal liver tissue architecture. Hazem et al. [52] observed that DAPA represent a viable approach to protect liver versus diabetes-induced steatohepatitis via suppressing oxidative stress, fibrosis progression and inflammation thus conserving hepatic functions and structure.

In conclusion, this research confirms that SGLT2 inhibition via DAPA and HDAC inhibition by SAHA decrease blood glucose level and protective in slowing the progression of liver damage in T2DM rats by improve IR and lipid profile and reduced tissue oxidative stress. The effect of DAPA was better than SAHA in decreasing fasting blood glucose level and improving lipid profile; while SAHA was better than DAPA in improving insulin secretion and IR and increased antioxidant and improvement of hepatic tissue damage. These findings suggest that selective SGLT2i and HDACi could be utilized in combination with other oral anti-diabetes drugs and insulin to further improve glycemic control and reverse organ damage in T2DM.

Authorship contribution

ZZA Resources, Conceptualization, Methodology, Writing – original draft, Preparation– review & editing. SIM Methodology, Resources, Writing – original draft, and editing. Both authors approved manuscript for publication.

Conflict of interest

Authors state that none of known financial conflicts or interpersonal ties that may have looked to have an impact on work described in this article.

Funding

This is a self-funded research.

Declarations Ethics approval

Research made after getting ethical approval from Ethical Committee of King Abdulaziz University, Jeddah, Saudi Arabia.

Consent for publication

Not applicable.

Competing interests

Authors declare that they had no competing interests.

Teo ZL, Tham Y-C, Yu M, Chee ML, Rim TH, Cheung N, et al. (2021) Global prevalence of diabetic retinopathy and projection of burden through 2045: systematic review and meta-analysis. Ophthalmology. 128(11):1580–91. https://doi.org/10.1016/j.ophtha.2021.04.027
Salem IS, El Mageed NMA, El Rahman H (2022) Effect Of Green Coffee Bean Extract On Rats Suffering From Diabetes. Journal of Pharmaceutical Negative Results. 13(9):8130–43. https://doi.org/10.47750/pnr.2022.13.S09.950
Kim H, Lee DS, An TH, Park H-J, Kim WK, Bae K-H, et al. (2021) Metabolic spectrum of liver failure in type 2 diabetes and obesity: from NAFLD to NASH to HCC. International journal of molecular sciences. 22(9):4495. https://doi.org/10.3390/ijms22094495
Khan S, Jena G, Tikoo K, Kumar V (2015) Valproate attenuates the proteinuria, podocyte and renal injury by facilitating autophagy and inactivation of NF-κB/iNOS signaling in diabetic rat. Biochimie. 110:1–16. https://doi.org/10.1016/j.biochi.2014.12.015
Khan S, Kowluru A (2018) CD36 mediates lipid accumulation in pancreatic beta cells under the duress of glucolipotoxic conditions: Novel roles of lysine deacetylases. Biochemical and biophysical research communications. 495(3):2221–6. https://doi.org/10.1016/j.bbrc.2017.12.111
Hadden MJ, Advani A (2018) Histone Deacetylase Inhibitors and Diabetic Kidney Disease. International journal of molecular sciences. 19(9):2630. https://doi.org/10.3390/ijms19092630
Makkar R, Behl T, Arora S (2020) Role of HDAC inhibitors in diabetes mellitus. Current Research in Translational Medicine. 68(2):45–50. https://doi.org/10.1016/j.retram.2019.08.001
Chou DH, Holson EB, Wagner FF, Tang AJ, Maglathlin RL, Lewis TA, et al. (2012) Inhibition of histone deacetylase 3 protects beta cells from cytokine-induced apoptosis. Chemistry & biology. 19(6):669–73. https://doi.org/10.1016/j.chembiol.2012.05.010
Lundh M, Christensen DP, Damgaard Nielsen M, Richardson SJ, Dahllöf MS, Skovgaard T, et al. (2012) Histone deacetylases 1 and 3 but not 2 mediate cytokine-induced beta cell apoptosis in INS-1 cells and dispersed primary islets from rats and are differentially regulated in the islets of type 1 diabetic children. Diabetologia. 55(9):2421–31. https://doi.org/10.1007/s00125-012-2615-0
Grant S, Easley C, Kirkpatrick P (2007) Vorinostat. Nat Rev Drug Discov. 6(1):21–2. https://doi.org/10.1038/nrd2227
Cabrera SM, Colvin SC, Tersey SA, Maier B, Nadler JL, Mirmira RG (2013) Effects of combination therapy with dipeptidyl peptidase-IV and histone deacetylase inhibitors in the non-obese diabetic mouse model of type 1 diabetes. Clin Exp Immunol. 172(3):375–82. https://doi.org/10.1111/cei.12068
Kashiwagi A, Maegawa H (2017) Metabolic and hemodynamic effects of sodium-dependent glucose cotransporter 2 inhibitors on cardio-renal protection in the treatment of patients with type 2 diabetes mellitus. Journal of diabetes investigation. 8(4):416–27. https://doi.org/10.1111/jdi.12644
Harris K, Nealy KL (2020) Cardiorenal Outcomes in Type 2 Diabetes. Consensus statement. 43:S98-S110.
Scheen AJ (2019) Beneficial effects of SGLT2 inhibitors on fatty liver in type 2 diabetes: A common comorbidity associated with severe complications. Diabetes & metabolism. 45(3):213–23. https://doi.org/10.1016/j.diabet.2019.01.008
Qiang S, Nakatsu Y, Seno Y, Fujishiro M, Sakoda H, Kushiyama A, et al. (2015) Treatment with the SGLT2 inhibitor luseogliflozin improves nonalcoholic steatohepatitis in a rodent model with diabetes mellitus. Diabetology & metabolic syndrome. 7:104. https://doi.org/10.1186/s13098-015-0102-8
Yokono M, Takasu T, Hayashizaki Y, Mitsuoka K, Kihara R, Muramatsu Y, et al. (2014) SGLT2 selective inhibitor ipragliflozin reduces body fat mass by increasing fatty acid oxidation in high-fat diet-induced obese rats. European journal of pharmacology. 727:66–74. https://doi.org/10.1016/j.ejphar.2014.01.040
Kapur A, O'Connor-Semmes R, Hussey EK, Dobbins RL, Tao W, Hompesch M, et al. (2013) First human dose-escalation study with remogliflozin etabonate, a selective inhibitor of the sodium-glucose transporter 2 (SGLT2), in healthy subjects and in subjects with type 2 diabetes mellitus. BMC pharmacology & toxicology. 14:26. https://doi.org/10.1186/2050-6511-14-26
Ying X, Chen X, Wang T, Zheng W, Chen L, Xu Y (2020) Possible osteoprotective effects of myricetin in STZ induced diabetic osteoporosis in rats. European journal of pharmacology. 866:172805. https://doi.org/10.1016/j.ejphar.2019.172805
Ulger TG, Cakiroglu FP (2020) The effects of onion (Allium cepa L.) dried by different heat treatments on plasma lipid profile and fasting blood glucose level in diabetic rats. Avicenna journal of phytomedicine. 10(4):325–33.
Mali VR, Ning R, Chen J, Yang XP, Xu J, Palaniyandi SS (2014) Impairment of aldehyde dehydrogenase-2 by 4-hydroxy-2-nonenal adduct formation and cardiomyocyte hypertrophy in mice fed a high-fat diet and injected with low-dose streptozotocin. Experimental Biology and Medicine. 239(5):610–8. https://doi.org/10.1177/1535370213520109
Colussi C, Berni R, Rosati J, Straino S, Vitale S, Spallotta F, et al. (2010) The histone deacetylase inhibitor suberoylanilide hydroxamic acid reduces cardiac arrhythmias in dystrophic mice. Cardiovascular Research. 87(1):73–82. https://doi.org/10.1093/cvr/cvq035
Arab HH, Fikry EM, Alsufyani SE, Ashour AM, El-Sheikh AAK, Darwish HW, et al. (2023) Stimulation of Autophagy by Dapagliflozin Mitigates Cadmium-Induced Testicular Dysfunction in Rats: The Role of AMPK/mTOR and SIRT1/Nrf2/HO-1 Pathways. Pharmaceuticals. 16(7):1006. https://doi.org/10.3390/ph16071006
Erdal GŞ, Karandere F, Mısıroğlu F, Tevetoğlu IÖ, Okuturlar Y, Koçoğlu H, et al. (2019) Assessment of relationship between serum magnesium and serum glucose levels and HOMA-IR in diabetic and prediabetic patients. Archives of Clinical and Experimental Medicine. 4(1):6–9. https://doi.org/10.25000/acem.451930
Alpdemir M, Alpdemir MF (2021) Comparison of Martin and Friedewald equation for estimated LDL-C in adults. Turk Kardiyoloji Dernegi Arsivi. 49(8):619. https://doi.org/10.5543/tkda.2021.90446
Bedi O, Aggarwal S, Trehanpati N, Ramakrishna G, Krishan P (2019) Molecular and Pathological Events Involved in the Pathogenesis of Diabetes-Associated Nonalcoholic Fatty Liver Disease. Journal of clinical and experimental hepatology. 9(5):607–18. https://doi.org/10.1016/j.jceh.2018.10.004
Zhu Y, Su Y, Zhang J, Zhang Y, Li Y, Han Y, et al. (2021) Astragaloside IV alleviates liver injury in type 2 diabetes due to promotion of AMPK/mTOR–mediated autophagy. Molecular medicine reports. 23(6). https://doi.org/10.3892/mmr.2021.12076
Wang K, Wang H, Liu Y, Shui W, Wang J, Cao P, et al. (2018) Dendrobium officinale polysaccharide attenuates type 2 diabetes mellitus via the regulation of PI3K/Akt-mediated glycogen synthesis and glucose metabolism. Journal of Functional Foods. 40:261–71.
Kelly WK, Richon VM, O'Connor O, Curley T, MacGregor-Curtelli B, Tong W, et al. (2003) Phase I clinical trial of histone deacetylase inhibitor: suberoylanilide hydroxamic acid administered intravenously. Clinical cancer research: an official journal of the American Association for Cancer Research. 9(10 Pt 1):3578–88. https://doi.org/10.1158/1078-0432.CCR-14-2556
Kelly WK, O'Connor OA, Krug LM, Chiao JH, Heaney M, Curley T, et al. (2005) Phase I study of an oral histone deacetylase inhibitor, suberoylanilide hydroxamic acid, in patients with advanced cancer. Journal of clinical oncology: official journal of the American Society of Clinical Oncology. 23(17):3923–31. https://doi.org/10.1200/jco.2005.14.167
Phrueksotsai S, Pinyopornpanish K, Euathrongchit J, Leerapun A, Phrommintikul A, Buranapin S, et al. (2021) The effects of dapagliflozin on hepatic and visceral fat in type 2 diabetes patients with non-alcoholic fatty liver disease. Journal of gastroenterology and hepatology. 36(10):2952–9. https://doi.org/10.1111/jgh.15580
Said Ahmed WM, Soliman A, Ahmed Amer AE, El Shahat RM, Amin MM, Taha RS, et al. (2023) Effect of dapagliflozin against NAFLD and dyslipidemia in type 2 diabetic albino rats: possible underlying mechanisms. European review for medical and pharmacological sciences. 27(17):8101–9. https://doi.org/10.26355/eurrev_202309_33570
Eriksson JW, Lundkvist P, Jansson PA, Johansson L, Kvarnström M, Moris L, et al. (2018) Effects of dapagliflozin and n-3 carboxylic acids on non-alcoholic fatty liver disease in people with type 2 diabetes: a double-blind randomised placebo-controlled study. Diabetologia. 61(9):1923–34. https://doi.org/10.1007/s00125-018-4675-2
Esterline RL, Vaag A, Oscarsson J, Vora J (2018) MECHANISMS IN ENDOCRINOLOGY: SGLT2 inhibitors: clinical benefits by restoration of normal diurnal metabolism? European journal of endocrinology. 178(4):R113-r25. https://doi.org/10.1530/eje-17-0832
Abdulwahab DA, El-Missiry MA, Shabana S, Othman AI, Amer ME (2021) Melatonin protects the heart and pancreas by improving glucose homeostasis, oxidative stress, inflammation and apoptosis in T2DM-induced rats. Heliyon. 7(3):e06474. https://doi.org/10.1016/j.heliyon.2021.e06474
Bocchi L, Motta BM, Savi M, Vilella R, Meraviglia V, Rizzi F, et al. (2019) The histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) restores cardiomyocyte contractility in a rat model of early diabetes. International journal of molecular sciences. 20(8):1873. https://doi.org/10.3390/ijms20081873
Silva G, Ferraresi C, de Almeida RT, Motta ML, Paixao T, Ottone VO, et al. (2020) Insulin resistance is improved in high-fat fed mice by photobiomodulation therapy at 630 nm. Journal of biophotonics. 13(3):e201960140. https://doi.org/10.1002/jbio.201960140
Elsharkawi I, Parambath D, Saber-Ayad M, Khan AA, El-Serafi AT (2020) Exploring the effect of epigenetic modifiers on developing insulin-secreting cells. Human cell. 33(1):1–9. https://doi.org/10.1007/s13577-019-00292-y
Dewanjee S, Vallamkondu J, Kalra RS, Chakraborty P, Gangopadhyay M, Sahu R, et al. (2021) The emerging role of HDACs: pathology and therapeutic targets in diabetes mellitus. Cells. 10(6):1340. https://doi.org/10.3390/cells10061340
Tang L, Wu Y, Tian M, Sjöström CD, Johansson U, Peng XR, et al. (2017) Dapagliflozin slows the progression of the renal and liver fibrosis associated with type 2 diabetes. American journal of physiology Endocrinology and metabolism. 313(5):E563-e76. https://doi.org/10.1152/ajpendo.00086.2017
Joannides CN, Mangiafico SP, Waters MF, Lamont BJ, Andrikopoulos S (2017) Dapagliflozin improves insulin resistance and glucose intolerance in a novel transgenic rat model of chronic glucose overproduction and glucose toxicity. Diabetes, obesity & metabolism. 19(8):1135–46. https://doi.org/10.1111/dom.12923
Millar PJ, Pathak V, Moffett RC, Pathak NM, Bjourson AJ, O'Kane MJ, et al. (2016) Beneficial metabolic actions of a stable GIP agonist following pre-treatment with a SGLT2 inhibitor in high fat fed diabetic mice. Molecular and cellular endocrinology. 420:37–45. https://doi.org/10.1016/j.mce.2015.11.019
Zhang Y, Geng Z, XU P, Gao X (2013) Effect of suberoylanilide hydroxamic acid on liver injury induced by lethal hemorrhagic shock in rats first entering high altitude. Chinese Journal of Anesthesiology.477 – 80.
Zhao Y, Zhou P, Liu B, Bambakidis T, Mazitschek R, Alam HB, et al. (2015) Protective effect of suberoylanilide hydroxamic acid against lipopolysaccharide-induced liver damage in rodents. Journal of Surgical Research. 194(2):544–50. https://doi.org/10.1016/j.jss.2014.10.056
Wang Y, Zhao L, Jiao FZ, Zhang WB, Chen Q, Gong ZJ (2018) Histone deacetylase inhibitor suberoylanilide hydroxamic acid alleviates liver fibrosis by suppressing the transforming growth factor-β1 signal pathway. Hepatobiliary & pancreatic diseases international: HBPD INT. 17(5):423–9. https://doi.org/10.1016/j.hbpd.2018.09.013
Alhaddad A, Abdel Kawy H, Ramadan W, Abbas A (2019) Hepatoprotective Effects of Suberoylanilide Hydroxamic Acid (A Histone Deacetylase Inhibitor) in Immunological Liver Injury in Rats. Journal of Pharmaceutical Research International. 30(3):1–15. https://doi.org/10.9734/jpri/2019/v30i330271
Leng W, Wu M, Pan H, Lei X, Chen L, Wu Q, et al. (2019) The SGLT2 inhibitor dapagliflozin attenuates the activity of ROS-NLRP3 inflammasome axis in steatohepatitis with diabetes mellitus. Annals of translational medicine. 7(18):429. https://doi.org/10.21037/atm.2019.09.03
Finan B, Yang B, Ottaway N, Smiley DL, Ma T, Clemmensen C, et al. (2015) A rationally designed monomeric peptide triagonist corrects obesity and diabetes in rodents. Nature medicine. 21(1):27–36. https://doi.org/10.1038/nm.3761
Ito D, Shimizu S, Inoue K, Saito D, Yanagisawa M, Inukai K, et al. (2017) Comparison of Ipragliflozin and Pioglitazone Effects on Nonalcoholic Fatty Liver Disease in Patients With Type 2 Diabetes: A Randomized, 24-Week, Open-Label, Active-Controlled Trial. Diabetes care. 40(10):1364–72. https://doi.org/10.2337/dc17-0518
Leoni F, Zaliani A, Bertolini G, Porro G, Pagani P, Pozzi P, et al. (2002) The antitumor histone deacetylase inhibitor suberoylanilide hydroxamic acid exhibits antiinflammatory properties via suppression of cytokines. Proceedings of the National Academy of Sciences. 99(5):2995–3000. https://doi.org/10.1073/pnas.052702999
Wu Q, Cheng Z, Zhu J, Xu W, Peng X, Chen C, et al. (2015) Suberoylanilide hydroxamic acid treatment reveals crosstalks among proteome, ubiquitylome and acetylome in non-small cell lung cancer A549 cell line. Scientific reports. 5(1):9520. https://doi.org/10.1038/srep09520
Hsi LC, Xi X, Lotan R, Shureiqi I, Lippman SM (2004) The histone deacetylase inhibitor suberoylanilide hydroxamic acid induces apoptosis via induction of 15-lipoxygenase-1 in colorectal cancer cells. Cancer research. 64(23):8778–81. https://doi.org/10.1158/0008-5472.CAN-04-1867
Hazem RM, Ibrahim AZ, Ali DA, Moustafa YM (2022) Dapagliflozin improves steatohepatitis in diabetic rats via inhibition of oxidative stress and inflammation. International immunopharmacology. 104:108503. https://doi.org/10.1016/j.intimp.2021.108503
Musso G, Gambino R, Cassader M, Pagano G (2012) A novel approach to control hyperglycemia in type 2 diabetes: sodium glucose co-transport (SGLT) inhibitors: systematic review and meta-analysis of randomized trials. Annals of medicine. 44(4):375–93. https://doi.org/10.3109/07853890.2011.560181
Yuan T, Yang T, Chen H, Fu D, Hu Y, Wang J, et al. (2019) New insights into oxidative stress and inflammation during diabetes mellitus-accelerated atherosclerosis. Redox Biol. 20:247–60. https://doi.org/10.1016/j.redox.2018.09.025
Tanbek K, Ozerol E, Yilmaz U, Yilmaz N, Gul M, Colak C (2022) Alpha lipoic acid decreases neuronal damage on brain tissue of STZ-induced diabetic rats. Physiology & behavior. 248:113727. https://doi.org/10.1016/j.physbeh.2022.113727
Ağgül AG, Gür F, Gülaboğlu M (2021) Streptozotocin-Induced Oxidative Stress in Rats: The Protective Role of Olive Leaf Extract. Bulletin of the Korean Chemical Society. 42(2):180–7. https://doi.org/10.1002/bkcs.12157
Bakhdar FA, Magadmi RM, El-Kordy EA, Alamri AS (2023) Effect of histone deacetylase inhibitor (vorinostat) on new-onset diabetes induced by tacrolimus. Journal of Taibah University Medical Sciences. 18(1):9–18. https://doi.org/10.1016/j.jtumed.2022.07.004
Kashiwagi A (2001) Complications of diabetes mellitus and oxidative stress. Japan Medical Association Journal. 44(12):521–8.
Al-Ani I, Al-Mishadani N, Muslih RK, Hamoodi SR (2009) Histological liver changes in streptozotocin induced diabetic mice. Med J Malaysia. 8(1).
Rodríguez V, Plavnik L, de Talamoni NT (2018) Naringin attenuates liver damage in streptozotocin-induced diabetic rats. Biomedicine & Pharmacotherapy. 105:95–102.
Ugwu M, Umar I, Utu-Baku A, Dasofunjo K, Ukpanukpong R, Yakubu O, et al. (2013) Antioxidant status and organ function in streptozotocin-induced diabetic rats treated with aqueous, methanolic and petroleum ether extracts of Ocimum basilicum leaf. Journal of Applied Pharmaceutical Science. 3(4):S75.
Chen Y, Du J, Zhao YT, Zhang L, Lv G, Zhuang S, et al. (2015) Histone deacetylase (HDAC) inhibition improves myocardial function and prevents cardiac remodeling in diabetic mice. Cardiovascular diabetology. 14:1–13.
Bocchi L, Motta BM, Savi M, Vilella R, Meraviglia V, Rizzi F, et al. (2019) The Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid (SAHA) Restores Cardiomyocyte Contractility in a Rat Model of Early Diabetes. International journal of molecular sciences. 20(8). https://doi.org/10.3390/ijms20081873
Santos CX, Anilkumar N, Zhang M, Brewer AC, Shah AM (2011) Redox signaling in cardiac myocytes. Free radical biology & medicine. 50(7):777–93. https://doi.org/10.1016/j.freeradbiomed.2011.01.003

No competing interests reported.

Download PDF

Version 1

posted

You are reading this latest preprint version

Protective effects of Suberoylanilide hydroxamic acid and Dapagliflozin administration on liver of diabetic rats

Status:

Version 1

Abstract

Background

Methods

Results

Conclusions

Figures

Introduction

Material and method

Animals

Chemicals and drugs

Experimental design and induction of T2DM

Hepatic tissue homogenate preparation

Biochemical Measurements

Histological examination

Statistical Analysis

Results

Biological results

Glycemic control results

Liver function test results

Lipid profile results

Oxidative stress results

Histology results

Discussion

Conclusions

Declarations

References

Additional Declarations

Status:

Version 1