Virgin Coconut oil is known for its health and therapeutic benefits and is exploited because of its ability to scavenge free radicals. This study aimed to evaluate the impact of virgin coconut oil on dolutegravir-induced organ toxicity in Wistar albino rats. Fifteen healthy Wistar Albino rats were allotted into five groups of three animals each. Animals in the negative control group were administered normal saline, while those in group two had dolutegravir (10 mg/kg oral) daily for 28 days. Rats in groups three, four, and five received dolutegravir (10 mg/kg) plus virgin coconut oil (5, 10, and 15 ml/kg), respectively. All treatments were administered once daily for twenty-eight days. Twenty-four hours after the last dose, the animals were sacrificed, and blood samples were collected for hematological, lipid profile, and biochemical analysis, while the liver of each animal was harvested and processed for histological analysis. Administration of virgin coconut oil in the presence of dolutegravir caused an initial insignificant decrease (P > 0.05) in alanine aminotransferase and aspartate aminotransferase levels compared to Dolutegravir alone. We found that White and Red blood cell counts increased in the groups receiving virgin coconut oil compared to animals in the control group (dolutegravir only). Virgin coconut oil, in the presence of dolutegravir, decreased levels of triglycerides and increased levels of high-density lipoproteins. Serum creatinine and urea were observed to decrease in the group co-administered dolutegravir and virgin coconut oil. The oil showed protective effects against dolutegravir-induced histological changes in the liver tissue of rats. It can be concluded that virgin coconut oil protected the liver as well as the kidney from dolutegravir-induced toxicity.
Article
Virgin Coconut Oil Protects Against Dolutegravir-induced Toxicity in Wistar Albino Rats
https://doi.org/10.21203/rs.3.rs-3991126/v1
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Virgin coconut oil
dolutegravir
hepatotoxicity
nephrotoxicity
oxidative stress
The liver is the largest organ in the human body and is important for metabolic processes, including glycogen storage, decomposition of red blood cells, plasma protein synthesis, and detoxification [1]. Due to the peculiarity of its function, this organ is continuously and variedly exposed to xenobiotics, environmental pollutants, and chemotherapeutic agents [2]. If the natural protective mechanisms of the liver are overpowered during all such processes, hepatic injury results.
Hepatotoxicity refers to liver damage caused by exposure to certain chemicals. Some medicinal substances, when consumed in excessive amounts or occasionally even within prescribed therapeutic limits, can harm the liver. Additionally, various chemical agents like those utilized in laboratories and industries, as well as natural substances such as microcystins and herbal remedies, can induce hepatotoxicity [3]. Drug-induced liver injury is responsible for 5% of all hospital admissions and 50% of all acute liver failures. Over 75% percent of cases of idiosyncratic drug reactions result in liver transplantation or death [4].
Despite the significant advancements in modern medicine, drugs that effectively protect the liver or contribute to the regeneration of hepatic cells, either by directly or indirectly stimulating liver function, are lacking [5]. This underscores the complexity of addressing hepatic health and highlights the need for further research and innovative approaches in the quest for effective liver protective and regenerative therapies.
The link between antioxidant and hepatoprotective mechanisms has been established [5] [10]. Virgin coconut oil (VCO), extracted from the nuts of the palm tree Cocos nucifera, has numerous medicinal properties, such as antioxidant [6], antimicrobial, antiviral [7][8], antilipidemic and antithrombotic [9] activities. Reportedly, Coconut Oil treats and prevents thyroid diseases, promotes weight loss, and improves digestion. Furthermore, VCO is capable of increasing antioxidant enzyme activity and reducing lipids [9].
Dolutegravir, classified as an integrase strand transfer inhibitor (INSTI), is a pivotal component within the regimens for managing HIV infection. Its remarkable efficacy, high tolerability, and rare occurrences of drug interactions render the co-formulation of dolutegravir with two nucleotide reverse-transcriptase inhibitors a compelling treatment choice [11]. Potential adverse effects include allergic responses and hepatic complications [12]. The administration of INSTIs alone has been linked to adverse effects such as insomnia accompanied by neuropsychiatric reactions [13], as well as diarrhea, vomiting, allergic reactions, and elevated liver enzymes [11] [14]. Decreased creatinine clearance has been reported. In vitro experiments revealed that Dolutegravir induces cellular shrinkage and the rearrangement of phospholipids within cell membranes. These effects are thought to be associated with oxidative stress caused by Dolutegravir [15]. Although the hepatoprotective role of natural products has been investigated, the specific role of VCO on dolutegravir-induced hepatotoxicity has remained unexplored. Therefore, this study aims to evaluate the protective effects of VCO on dolutegravir-induced hepatotoxicity, as well as other organ toxicities.
2.1 Preparation of Virgin Coconut Oil
The oil was extracted from coconut obtained from the local market. The coconut flesh was freshly grated, placed in a cheesecloth, and wrapped. The cheesecloth was then compressed manually to extract the coconut milk from the flesh until no more liquid was released. Subsequently, the strained coconut milk was transferred to a stainless pot, and the oil was extracted by heating at 100–120°C for approximately 60 min until the water was evaporated entirely. After heating, the resultant oil was separated from the pertinacious residue by filtering through a muslin cloth, and the remaining residue was further heated to extract more oil [16]. Finally, the oil was stored in a bottle.
2.2 Experimental animals
Female Wistar rats (females) weighing 100–150 g were obtained from the National Veterinary Research Institute (NVRI), Vom, Nigeria. The animals were housed at 25ºC with a 12-hour day-night cycle, allowed to acclimatize for fourteen days before the experiment, and were fed and allowed water ad libitum. The study protocol was approved by the ethics committee of the Animal Experimental Unit of the University of Jos, Nigeria and reported in accordance with the ARRIVE guidelines. Furthermore, all methods were performed in accordance with the relevant guidelines and regulations.
2.3 Experimental Procedure
2.3.1 Acute toxicity test
Acute toxicity (LD50) determination was performed using the modified Lorke’s method [17]. The evaluation was done in two phases using twelve animals.
In phase one, three groups comprising three rats each were orally administered 10, 100, and 1000 mg/kg body weight of dolutegravir, respectively, and observed for 24 h for mortality.
In the second phase, three groups comprising one animal each were orally administered 1600, 2900, and 5000 mg/kg doses of dolutegravir, respectively. They were observed for 24 h for mortality.
The LD50 was calculated using the formula:
LD50 =\(\sqrt{D0}\times D100 (\text{L}\text{o}\text{r}\text{k}\text{e}, 1983)\)
where D0 = Highest dose that caused no mortality
D100 = Lowest dose that caused mortality.
2.3.1 Drug administration
Animals were divided into five groups of 3 rats each, and the administration of the drug was done as follows;
Group 1 (Control): Animals in this group were administered only water and food; Group 2: Animals in this group were administered dolutegravir only (10 mg/kg); Group 3: Animals in this group were administered dolutegravir (10 mg/kg) and virgin coconut oil (5 ml/kg); Group 4: Animals in this group were administered dolutegravir (10 mg/kg) and virgin coconut oil (10 ml/kg); Group 5: Animals in this group were administered dolutegravir (10 mg/kg) and virgin coconut oil (15 ml/kg).
2.3.3 Biochemical, haematological and lipid profile analysis
After drug administration, blood samples were collected from each rat via the jugular artery under anesthesia into plain and EDTA sample bottles for biochemical and hematological analyses, respectively. Blood collected into plain sample bottles were allowed to clot at room temperature (250C) and centrifuged to obtain the serum. The sera were analyzed using a Cobas c111 analyzer (Roche Diagnostics (Schweiz) AG, Switzerland) to assess the hematological parameters (white blood cells, red blood cells, hemoglobin, and packed cell volume).
Lipid profile parameters, including total cholesterol, triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL), total protein, albumin, and bilirubin, were analyzed using Cobas c111.
Furthermore, using Cobas c111, the liver and kidney functions were determined by assessing the levels of Alanine aminotransferase (ALT), Aspartate aminotransferase (AST) and Alkaline phosphatase (ALP) for liver, and urea and creatinine for the kidney.
2.3.4 Histological analysis
Rats were humanely sacrificed under inhaled chloroform anesthesia, laparostomized and the liver of each animal was removed and placed in an organ bottle containing 10% formalin.
The procedure followed was based on the methodology outlined by Saalu et al. [18]. In brief, the organs were sliced into approximately 0.5 cm thick slabs and immersed in 10% formal saline solution for one day. Subsequently, they were moved to 70% alcohol to initiate tissue dehydration. Following this, the tissues underwent sequential immersion in 90% alcohol and 10% chloroform for varying durations. They were then placed in two successive changes of molten paraffin wax for 20 minutes each at 57°C in an oven. Subsequently, 5 mm thick serial sections were obtained from the solid tissue blocks and stained using hematoxylin and eosin. The stained sections were then treated with a mixture of equal parts xylene and alcohol before undergoing clearance in xylene and subsequent oven-drying. Photomicrographs were captured using a color digital camera attached to a light microscope.
2.4 Statistical analysis
Data were analyzed by one-way ANOVA followed by Turkey’s multiple comparison tests using Graph pad Prism 5 (Graph pad Software Inc. CA). Results were expressed as mean SEM, and values were considered significant at P < 0.05.
3.1 Determination of acute toxicity (LD50)
The acute toxicity test study of the dolutegravir showed that the drug was safe up to 1,254.9 mg/kg, with no mortality observed at that concentration (Table 1).
Table 1: Determination of acute toxicity (LD50)
LD50 = 1254.9 mg/kg
3.2 Effect of virgin coconut oil and dolutegravir treatment on some hematological parameters
There was an insignificant increase in the white and red blood cell levels in the test animals (Table 2).
|
Parameters |
Grp 1 |
Grp 2 |
Grp 3 |
Grp 4 |
Grp 5 |
|---|---|---|---|---|---|
|
WBC (X103 /µl) |
5.07 ± 1.38 |
7.43 ± 2.09 |
4.40 ± 1.99 |
5.53 ± 0.32 |
12.87 ± 3.52 |
|
RBC (X 103/ µl) |
7.95 ± 1.12 |
6.55 ± 1.84 |
8.90 ± 0.44 |
4.23 ± 1.13 |
8.37 ± 0.19 |
|
HB (g/dl) |
15.07 ± 2.40 |
14.57 ± 2.03 |
14.17 ± 4.20 |
8.33 ± 2.18 |
15.07 ± 0.88 |
|
PCV (%) |
46.57 ± 5.35 |
50.37 ± 4.87 |
60.47 ± 1.07 |
33.97 ± 9.55 |
51.3 ± 1.30 |
|
LYMP (%) |
54.00 ± 9.82 |
60.53 ± 42.07 |
42.07 ± 3.98 |
51.53 ± 2.62 |
45.60 ± 4.44 |
|
MONO (%) |
9.43 ± 1.83 |
5.23 ± 0.83 |
7.37 ± 0.55 |
9.00 ± 4.02 |
9.90 ± 3.00 |
|
NEUT (%) |
36.57 ± 9.45 |
37.80 ± 3.92 |
50.57 ± 4.54 |
39.47 ± 1.43 |
44.45 ± 1.65 |
|
PLT (X 103/ µl) |
462.70 ± 28.85 |
778.00 ± 271.40 |
725.30 ± 97.50 |
384.70 ± 139.40 |
813.30 ± 128.10 |
| Keys: Grp 1, Control group; Grp 2, Dolutegravir 10 mg/kg; Grp 3, 4 and 5, Dolutegravir 10 mg/kg + Virgin Coconut Oil 5, 10 and 15 ml/kg, respectively; WBC, White blood count; RBC, Red blood count; HB, Hemoglobin; PCV, Packed cell volume; LYMP, Lymphocytes; MONO, Monocytes; NUET, Neutrophils; PLT, Platelets. | |||||
3.3 Effect of virgin coconut oil on dolutegravir-induced kidney injury
A significant (P < 0.05) decrease in the levels of urea was observed for animals administered the test drug and VCO (5 and 10 ml/kg) compared to the group administered the drug alone (Table 3).
|
Parameters |
Grp 1 |
Grp 2 |
Grp 3 |
Grp 4 |
Grp 5 |
|---|---|---|---|---|---|
|
Creatinine (mg/dl) |
0.20 ± 0.05 |
0.23 ± 0.07 |
0.23 ± 0.03 |
0.20 ± 0.00 |
0.17 ± 0.03 |
|
Urea (mmol/l) |
5.10 ± 0.37 |
5.87 ± 0.65 |
3.29 ± 0.14* |
3.24 ± 0.62* |
5.70 ± 0.34 |
| Keys: *=P < 0.05; Results are expressed as Mean ± SEM. Grp 1, Control group; Grp 2, Dolutegravir 10 mg/kg; Grp 3, 4 and 5, Dolutegravir 10 mg/kg + Virgin Coconut Oil 5, 10 and 15 ml/kg respectively. | |||||
3.4 Effect of virgin coconut oil and dolutegravir treatment on some lipid parameters
A significant increase (P < 0.05) in the levels of Triglycerides was observed for animals administered test drugs and 15 ml/kg of VCO (Table 4).
|
Parameters |
Grp 1 |
Grp 2 |
Grp 3 |
Grp 4 |
Grp 5 |
|---|---|---|---|---|---|
|
TG (mmol/l) |
0.92 ± 0.17 |
0.85 ± 0.08 |
0.56 ± 0.01 |
1.00 ± 0.13 |
1.87 ± 0.22* |
|
CHOL (mmol/l) |
1.67 ± 0.12 |
1.48 ± 0.02 |
1.70 ± 0.04 |
1.93 ± 0.02 |
1.74 ± 0.09 |
|
HDL (mmol/l) |
0.59 ± 0.08 |
0.50 ± 0.07 |
0.55 ± 0.05 |
0.59 ± 0.11 |
0.40 ± 0.04 |
| *=P < 0.05 | |||||
| TG, Triglycerides; CHOL, Cholesterol; HDL, High-density lipoprotein. | |||||
| Grp 1; Control group; Grp 2, Dolutegravir 10 mg/kg; Grp 3, 4 and 5, Dolutegravir 10 mg/kg + Virgin Coconut Oil 5, 10 and 15 ml/kg respectively. | |||||
3.5 Effect of virgin coconut oil on dolutegravir-induced liver injury
Animals treated with dolutegravir and 15 ml/kg of VCO showed a significant increase (P < 0.05) in the levels of ALT and ALP and a significant decrease in TP levels compared to the other groups. Animals treated with test drug and 10 ml/kg VCO also showed a significant increase (P < 0.05) in ALP levels (Table 5).
|
Parameters |
Grp 1 |
Grp 2 |
Grp 3 |
Grp 4 |
Grp 5 |
|---|---|---|---|---|---|
|
ALT (IU/L) |
60.30 ± 6.61 |
55.93 ± 11.94 |
55.93 ± 5.23 |
76.50 ± 8.41 |
110.00 ± 9.62* |
|
AST (IU/L) |
210.90 ± 31.18 |
222.60 ± 36.94 |
176.90 ± 7.44 |
176.10 ± 14.30 |
197.30 ± 38.50 |
|
ALP (IU/L) |
386.3 ± 69.42 |
375.00 ± 106.20 |
504.70 ± 45.92 |
835.70 ± 94.27* |
1093.00 ± 144.74* |
|
T BIL (µmol/l) |
21.69 ± 9.80 |
27.77 ± 6.94 |
23.87 ± 4.72 |
23.30 ± 6.65 |
29.20 ± 9.43 |
|
T P (g/dl) |
6.97 ± 0.26 |
6.40 ± 0.05 |
6.07 ± 0.19 |
6.07 ± 0.28 |
5.93 ± 0.20* |
| *P < 0.05 | |||||
| Results are expressed as Mean ± SEM. Grp 1 = Normal control. Grp 2 = Dolutegravir 10 mg/kg, Grp 3, 4 and 5 = Dolutegravir 10 mg/kg + Virgin Coconut Oil 5, 10 and 15 ml/kg, respectively. | |||||
| TBIL, Total bilirubin; ALT, alanine transferase; ALP, Alkaline phosphatase; TP, Total protein; AST, Aspartate aminotransferase | |||||
3.6 Histology of the rat liver showing the effect of dolutegravir and virgin coconut oil administration
Figure 1A-E shows the histology of the liver before and after the administration of dolutegravir. After dolutegravir administration, the central vein became congested within the sinusoids. The level of liver damage decreased in the VCO + dolutegravir group (Fig. 1C-E).
In this study, we presented the biochemical and histological changes that resulted from dolutegravir and VCO treatment.
The diagnosis of liver and kidney disease is usually predicted by the changes in biochemical markers of injury. AST is a specific maker for liver injury; thus, it is elevated in the serum after an injury or during pathology. Dolutegravir has been reported to cause organ toxicity, especially to the liver [19]. This may be due to the fact that hepatic glutathione levels decrease during periods of metabolic stress, leading to the elevation of makers of organ injury [20].
Several studies have demonstrated that an increase in serum enzyme levels is a signal of an underlying pathological process [21]. It has also been reported that an increase in the serum levels of certain enzymes directly indicates the major pathologic changes in cell membrane permeability or hepatic cell rupture [22]. A similar observation was made in this present study following the administration of dolutegravir. Our findings revealed that after the administration of VCO to animals challenged with Dolutegravir, an initial insignificant decrease (P > 0.05) in the levels of ALT and AST compared to the controls was observed. This is an indication of the reported protective property of virgin coconut oil [19]. These authors reported insignificant differences (P > 0.05) in the activities of serum aminotransferases, which is consistent with the findings of the present study. Similarly, the decreased TP concentration observed in this present study is consistent with that reported by Moshira et al. [23].
Coconut water has been reported to have protective and regenerative effects on the kidney [24–25]. The rise in serum creatinine and urea levels observed in the current study among the group administered dolutegravir signifies its potential to induce kidney damage or dysfunction. If the kidney’s ability to filter urine is deficient, blood levels of creatinine rise. According to Hafeez et al. [26], creatinine levels in blood and urine may be used to calculate creatinine clearance (CrCl), which is a reflection of the glomerular filtration rate. Creatinine clearance test is important in evaluating the progression of renal disease, as well as in diagnosing renal dysfunction. A decrease in the kidney’s ability to clear creatinine is, therefore, an indication that there is a reduction in the glomerular filtration rate. The most consistently observed biochemical effect of dolutegravir on the kidney is creatinine elevation [27]. This was observed in the present study in animals administered dolutegravir when compared to animals in the control group. The antioxidant activity of virgin coconut oil was evident in its dose-dependent reduction in levels of creatinine, which is consistent with the findings by Intahphuak et al. [28]. Antioxidants play a critical role in hepatoprotection through diverse mechanisms. By neutralizing reactive oxygen species, antioxidants prevent oxidative stress-induced damage to liver cells and preserve cell membrane integrity. They enhance enzymatic defense systems, regulate inflammatory responses, and support detoxification processes, contributing to overall liver health. Moreover, antioxidants promote liver cell regeneration and modulate apoptosis, preventing excessive cell death. This multifaceted approach underscores the importance of antioxidants in safeguarding the liver from various stressors and highlights their therapeutic potential in preventing or ameliorating liver disorders [29].
Cholesterol is important to the body as a component of membranes and for the biosynthesis of steroid hormones. The elevation in total cholesterol and triglycerides observed with protease Inhibitors is absent with dolutegravir [27]. This was observed in the present study in the group administered Dolutegravir. VCO in the presence of dolutegravir, however, decreased the levels of triglycerides and increased that of high-density lipoproteins (HDL), although insignificant (P > 0.05). This was particularly evident at the lower doses. The present result particularly showed that there was no significant difference in the serum HDL levels across the treatments, consistent with the findings of [30]. Notably, however, in the present study, there was an insignificant increase in cholesterol levels in the test groups compared to the control group (P > 0.05). This observation is consistent with the report by Vasudevan [31], where it was demonstrated that virgin coconut oil contains a minimal amount of cholesterol.
The WBC and RBC count showed an increasing trend in the groups receiving VCO compared to animals in the control group. This is consistent with the findings of Senin et al. [32].
Furthermore, the histology showed that VCO had protective effects against dolutegravir-induced histological changes in the hepatocytes of Wistar rats. This protective effect of VCO was also reported by [32]. This was evidenced by the reduction in the damage to the liver architecture in groups receiving both DTG and VCO as compared to those receiving only dolutegravir. These histological findings were further supported by the progressive reduction in necrosis of the hepatocytes as the concentration of VCO increased (Fig. 1C, D, and E).
Although this study demonstrated the protective effect of VCO on the liver and kidney against dolutegravir-induced injury, the mechanism of such effects needs to be explored in detail. These findings provide preliminary evidence for the protective effect of VCO. Clinical studies need to be conducted to ascertain the effectiveness of VCO in patients with HIV taking dolutegravir and other similar drugs. Additionally, VCO may be considered by nutritionists as a supplement for patients with hyperlipidemia.
Author Contribution
K.I.A. conceptualized and designed the study protocol and performed the statistical analysis; S.P.D. was involved with animal care and drug administration; L.A. and M.L.L. supervised the research and wrote the first draft of the manuscript. All authors equally worked on successive drafts of the manuscript until the final copy was produced.
Acknowledgment
The authors express gratitude to the Head of the Department of Pharmacology and Toxicology, University of Jos, for providing access to the department's laboratory facilities. Special appreciation is extended to Mr. Bulus Diyen and Mr. Sunday Azi, staff of the Department of Pharmacology & Toxicology Laboratory, for their valuable technical support. Additionally, sincere thanks are extended to Mr. Sale Gotom of the Histology Unit of the Department of Anatomy, University of Jos, for his assistance in preparing the slides for the histopathological analysis.
- Opoku, A., Ndlovu, I., Terblanche, S., & Hutchings, A. In vivo hepatoprotective effects of Rhoicissus tridentata subsp. cuneifolia, a traditional Zulu medicinal plant, against CCl4-induced acute liver injury in rats. South African J. Bot., 73, 372–377 (2007).
- Ibrahim, M. et al. Hepatoprotective activity of Sapindus mukorossi and Rheum emodi extracts: in vitro and in vivo studies. World J. Gastroenterol. 14, 2566–2571 (2008).
- Patel, T., Roberts, L. R., Jones, B. A. & Gores, G. J. Dysregulation of apoptosis as a mechanism of liver disease: an overview. Semin. Liver. Dis. 18, 105–114 (1998).
- Ostapowicz, G. et al. Results of a prospective study of acute liver failure at 17 tertiary care centers in the United States. Ann. Intern. Med. 137, 947–954 (2002).
- Chattopadhyay, R. R. Possible mechanism of hepatoprotective activity of Azadirachta indica leaf extract: part II. J. Ethnopharmacol. 89, 217–219 (2003).
- Zeng, Y. Q. et al. Virgin coconut oil: A comprehensive review of antioxidant activity and mechanisms contributed by phenolic compounds. Crit. Rev. Food. Sci. Nutr. 64, 1052–1075 (2024).
- Bergsson, G., Arnfinnsson, J., Karlsson, S. M., Steingrímsson, O. & Thormar, H. In vitro inactivation of Chlamydia trachomatis by fatty acids and monoglycerides. Antimicrob. Agents Chemother. 42, 2290–2294 (1998).
- German, J. B. & Dillard, C. J. Saturated fats: what dietary intake. Am. J. Clin. Nutr. 80, 550–559 (2004).
- Nevin, K. G. & Rajamohan, T. Beneficial effects of virgin coconut oil on lipid parameters and in vitro LDL oxidation. Clin. Biochem. 37, 830–835 (2004).
- Pramyothin, P., Ngamtin, C., Poungshompoo, S. & Chaichantipyuth, C. Hepatoprotective activity of Phyllanthus amarus Schum. et. Thonn. extract in ethanol-treated rats: in vitro and in vivo studies. J. Ethnopharmacol. 114, 169–173 (2007).
- Kandel, C. E. & Walmsley, S. L. Dolutegravir - a review of the pharmacology, efficacy, and safety in the treatment of HIV. Drug Des. Devel. Ther. 9, 3547–3555 (2015).
- Blanco Arévalo, J. L. & Whitlock, G. G. Dolutegravir: an exciting new kid on the block. Expert. Opin. Pharmacother. 15, 573–582 (2014).
- WHO. Updated Recommedations on First- Line and Second-Line Antiretroviral Regimens and Poost-Exposure Prophylaxis and Recommendations on Early Infant Diagnosis of HIV: Interim Guidelines. Supplement to 2016 Consolidated Guidelines on the Use of Antiretroviral (2018).
- Daimari, R., Kwape, L., & Oyekunle, A. A.. Suicidal overdose of dolutegravir: A case report. Southern Afri. J. of HIV Medicine, 19, 1–3 (2018).
- Al Mamun Bhuyan, A., Signoretto, E., Bissinger, R. & Lang, F. Enhanced Eryptosis Following Exposure to Dolutegravir. Cell Physiol. Biochem. 39, 639–650 (2016).
- Srivastava, Y., Semwal, A.D., & Majumdar, A. Quantitative and qualitative analysis of bioactive components present in virgin coconut oil. Cogent Food & Agriculture, 2, (2016).
- Lorke, D. A new approach to practical acute toxicity testing. Arch. Toxicol. 54, 275–287 (1983).
- Saalu, L.C., Kpela, T., Benebo, A.S., Oyewopo, A.O., Anifowope, E.O., & Oguntola, J.A. The dose-dependent testiculoprotective and testiculotoxic potentials of Telfairia occidentalis Hook f. leaves extract in rat. International Journal of Applied Research in Natural Products, 3, 27–38 (2010).
- Otuechere, C. A., Madarikan, G., Simisola, T., Bankole, O. & Osho, A. Virgin coconut oil protects against liver damage in albino rats challenged with the anti-folate combination, trimethoprim-sulfamethoxazole. J. Basic Clin. Physiol. Pharmacol. 25, 249–253 (2014).
- Chen, Y., Dong, H., Thompson, D. C., Shertzer, H. G., Nebert, D. W. & Vasiliou, V. Glutathione defense mechanism in liver injury: insights from animal models. Food Chem. Toxicol. 60, 38–44 (2013).
- Feldstein, A. E. Novel insights into the pathophysiology of nonalcoholic fatty liver disease. Semin. Liver Dis. 30, 391–401 (2010).
- Contreras-Zentella, M. L. & Hernández-Muñoz, R. Is Liver Enzyme Release Really Associated with Cell Necrosis Induced by Oxidant Stress. Oxid. Med. Cell Longev. 2016, 3529149 (2016).
- ELAbasy, M., Abdelhady, D., Kamel, T., & Shukry, A. Ameliorative Effect of Coconut Oil on Hematological, Immunological and Serum Biochemical Parameters in Experimentally infected Rabbits. Alexandria Journal of Veterinary Sciences, 5, 36 (2016).
- Nwangwa, E. K., & Chukwuemeka, P. A. Regenerative effects of coconut water and coconut milk on the pancreatic B-cells and cyto architecture in alloxan-induced diabetic Wistar albino rats. A. J. Tropic. Med. & Public Health, 1(3), 137–146 (2011).
- Nwangwa, K. E. The Reno-Protective Effects of Coconut Water on the Kidneys of Diabetic Wistar Rats. International Journal of Health Science, 2, 1–4 (2012).
- Hafeez, A. R., Idrees, M. K. & Akhtar, S. F. Accuracy of GFR estimation formula in determination of glomerular filtration rate in kidney donors: Comparison with 24 h urine creatinine clearance. Saudi J. Kidney Dis. Transpl. 27, 320–325 (2016).
- Curtis, L. et al. Dolutegravir: clinical and laboratory safety in integrase inhibitor-naive patients. HIV Clin. Trials 15, 199–208 (2014).
- Intahphuak, S., Khonsung, P. & Panthong, A. Anti-inflammatory, analgesic, and antipyretic activities of virgin coconut oil. Pharm. Biol. 48, 151–157 (2010).
- Saha, P. et al. Role of Natural Phenolics in Hepatoprotection: A Mechanistic Review and Analysis of Regulatory Network of Associated Genes. Front Pharmacol 10, 509 (2019).
- Wachidah Yuiwarti, E.Y., Saraswati, T. R., & Kusdiyantini, E. Effect of VCO and olive oil on HDL, LDL, and cholesterol level of hyperglycemic Rattus Rattus Norvegicus. Journal of Physics: Conference Series, 1025, 012064 (2018)
- Vasudevan, D. M. Coconut oil and health controversy. International Journal of Health and Rehabilitation Sciences, 2, 157–164 (2013).
- Senin, M.M., Al-Ani, I.M., Mahmud, M.I.A.M., Muhammad, N., & Kasmuri, H.M. (2020). Protective Effect of Virgin Coconut Oil on Cyclophosphamide-induced Histological Changes in Lymphoid Tissues. IIUM Medical Journal Malaysia, 17, (2020).
No competing interests reported.