The goal of this study was to see how barley and barely β-Glucan extracts, which have been demonstrated to have antioxidant activity and anti-inflammatory properties, affected immunomodulation. To begin with, the antioxidant activity and chemical content of barley and barely β-glucan were evaluated. Barley has traditionally been used as a folk remedy for a range of ailments, including immune system support and antioxidant protection (46). Mikulajová et al. (47) discovered that it contains the flavonoid catechin, which has a strong antioxidant effect and contributes to antioxidant capacity.
According to the data, the protein content of barely β-glucan extract was found to be lower than that of barley extract. Because the single-step extraction method effectively separated them from the barley flour to obtain barely β-glucan, furthermore, the results of this study show that barley is the best source for β-glucan extraction due to a large amount of barely β-glucan present. Similar findings were also reported by Limberger-Bayer et al. (48). Our findings show that barely β-glucan extract has a higher level of antioxidant activity than barley extract, which has anti-inflammatory properties. In addition, the findings of the current study consistent with those of previous studies (49, 50).β-Glucans are a major component of the soluble dietary fibre β-(1,3 − 1,4)- d-glucan, which is isolated from barley. Furthermore, because of their wide range of biological activities, including lowering blood cholesterol levels, they are widely employed as supplements and food additives (50).
The effects of inorganic arsenic (Ar2O3) on inflammation and immunity in experimental rats were studied in the second part of this study. In previous research, arsenic-induced oxidative stress, inflammation, and a heat shock response have been described in the brain, liver, and immunological organs (2, 3, 4). Furthermore, arsenic-produced reactive oxygen species (ROS) react with polyunsaturated fatty acids, increasing lipid peroxidation. The imbalance between the formation of oxygen-derived radicals and the cellular antioxidant capacity caused by arsenic trioxide is a crucial factor in disease progression (2). Lymphocytic necrosis and depletion with the appearance of tangible body macrophages, this study looked at the negative effects of 5 mg/kg arsenic trioxide intraperitoneal injection in rats, which causes necrosis in spleen lymphocytic cells by increasing oxidative stress in the spleen and serum, and the subsequent amelioration by barley extract and barley β-glucan extract in adult male rats. In most cells, electron leakage from the mitochondrial electron transport chain is the main source of ROS (51). It has been suggested that an increase in reactive oxygen species (ROS) as a biochemical mediator of apoptosis can cause cell death (6).
Drug companies have recently moved to refocus their attention away from anti-inflammatory drugs, which have a long list of side effects. Natural extracts with immunosuppressive and anti-inflammatory properties are being investigated in depth.
The goal of this study was to see if barley β-glucan and barley extract could protect against the harmful effects of arsenic trioxide as an antioxidant agent. Furthermore, the current study found that arsenic may produce significant alterations in antioxidant activity markers and histological spleen tissues in experimental rats, suggesting that arsenic trioxide could increase oxidative stress. In arsenic-treated rats, the same results have been described (52, 32, 53). According to research (32, 54), MDA is a good indicator of endogenous lipid peroxidation. As for oxidative stress biomarkers, we assessed antioxidant enzyme activity detoxifying after ATO treatment. According to the findings, arsenic trioxide administration resulted in a significant increase in MDA (G2), indicating oxidative damage to the spleen, whereas anti-oxidative activities (GPx and CAT) were reduced in comparison to other groups. The antioxidant enzymes were strongly regulated by the administration of barley extracts, particularly barley β-glucan extracts. As a result, barley β-glucan and barley extracts increased antioxidant activity (CAT and GPx) and decreased MDA levels in the spleen tissue of rats, which is a key organ in the immunological response. This suggests that barley β-glucan has antioxidant properties and can restore the activity of ATO-affected enzymes. These findings support previous research (55, 56) that found barely β-glucan to have significant antioxidant activity and potentially protective benefits against cell damage.
Arsenic toxicity increases the expression and release of the proinflammatory cytokines interleukin (IL-6, IL-8, and IL-1β), the production of which is linked to the generation of reactive oxygen species (57). The administration of arsenic trioxide resulted in alterations in cytokine levels in the current investigation. TNF-α and IL-1β concentrations increased significantly in rats treated alone with ATO (G2). This conclusion is consistent with previous findings that ATO-induced oxidative stress likely promotes inflammation, and ROS overproduction causes increased expression of pro-inflammatory molecules such as tumour necrosis factor-α (TNF-α), inducible nitric oxide synthase, and nitric oxide synthase (58). A high concentration of arsenic produced TNF-α release from mononuclear cells and induced apoptosis in T cells, according to Yu et al. (59) and Liu et al. (60).
As a result of these effects, arsenic trioxide-induced oxidative stress damages the cellular membrane, causing inflammation and the production of pro-inflammatory cytokines. Furthermore, the findings indicate that barley extracts and barely β-glucan extracts have anti-inflammatory properties and decrease ATO-induced inflammation. The levels of cytokines (IL-1β, and α-TNF) were also higher in rats given only ATO (G2) than in the control group (G1). These parameters were lowered after treatment with barley and barely β-glucan extracts. The results might be attributed to the antioxidant and neutrophil inhibitory activities of β-glucan. In addition, β-Glucan's antioxidant effects and reduction of cytokines have also been demonstrated in other studies (61, 47). According to a recent study, α-TNF has a function in tumor cell death and is boosted by toxins. α-TNF can also be employed as a biomarker for the toxicity of test substances (62). According to results, Barley β-glucan can increase endotoxin clearance via scavenger receptors by lowering α-TNF generation. These findings are consistent with the information provided by (63, 64). Therefore, β-glucans, lignans, vanillic acid, and arabinoxylan are the major anti-inflammatory components in barley (12).
A weak immune system can cause infections, tumor growth, and immunodeficiency-related disorders. Immunoglobulins are a group of proteins that function as antibodies. To eliminate pathogens and neutralize toxins, they combine with antigens. (65). The immune system is made up of a variety of interdependent cell types that work together to protect the body from bacterial, parasite, fungal, and viral infections, as well as tumor cell growth. Many of these cell types have specific functions. B cell lymphocytes are responsible for antibody-mediated immunity (humoral immunity). They produce immunoglobulins (antibodies), which are proteins that bind to specific antigens (66). My research supports this theory, demonstrating that when ATO is used, the immun state deteriorates. Immunoglobulin levels (IgG, IgM) were shown to be significantly higher in rats given either barely β-glucan extracts or barley flour extracts, although the 80, 160 mg/kg β-glucan (G4, G5) and 2.8 g/kg barley extract (G8) were shown to be more beneficial in terms of all immunoglobulin concentrations. According to Chan et al. (67), β-glucans immediately enter the proximal small intestine and are captured by macrophages. They are then internalized and fragmented into smaller sized β-glucans. The small β-glucan fragments are then released by macrophages and taken up in the bloodstream by circulating granulocytes, monocytes, and dendritic cells. After that, the immunological response is triggered (67).
Several experimental studies have reported antibacterial, antifungal, antiviral, and antioxidant activities of β-glucans (68, 69, 70, 71). Barley β-glucans can regulate the immune responses and combine innate and adaptive immunity (12). In several investigations (68, 69, 70, 71), β-Glucans are immune stimulants that boost both natural and adaptive immunity.
In the current investigation, arsenic trioxide-induced rats had higher AST and ALT activity in their serum, which could indicate liver tissue injury. Increased levels of these liver biomarkers in the blood suggest morphological and functional damage to the liver (25). Furthermore, the increased serum transaminases (AST and ALT) after ATO treatment disrupted not only the synthesis of energetic macromolecules, which are required for several vital and metabolic functions, but also the detoxification processes (5), because barely β-glucan and barely extracts were effective in lowering these parameters. These findings corroborate the findings of Liu et al. (60), who found that ATO caused hepatotoxic and haematological alterations in rats. Previous studies have shown that barely β-glucan lowers oxidative damage and protects liver and kidney function (12), as well as that polyphenols present in barley, such as (+)-catechin, protocatechuate, and quercetin, have hepatoprotective characteristics (72). Similarly, serum creatinine and urea, both renal function markers, increased significantly in rats following ATO treatment, possibly indicating purine and pyrimidine breakdown and decreased kidney function. Many research looked at the effects of ATO on serum creatinine and urea levels in rats (73, 74). Barley is the cheapest cereal grain. As a result, the health benefits of dietary β-glucans may reach the poorest populations, such as patients with chronic renal disease and healthy people with systemic pro-aging or inflammatory disorders, raising the risk for kidney and vascular damage (73). Furthermore, uremic toxin produced by bacteria is reduced by barely β-glucan (12).
By improving the ability of macrophages, neutrophils, and natural killer cells to respond to and fight a variety of threats such as bacteria, viruses, fungi, and parasites, beta-1,3-glucans help the body's immune system defend against foreign invaders (75). In vivo, arsenic trioxide causes DNA strand breakage in mice's blood leukocytes (76).
According to these data, ATO reduced haemoglobin, red cell count, white cell count, neutrophils, lymphocytes, monocytes, and haematocrit while increasing plate counts and eosinophils. Arsenic trioxide toxification and increased oxidative stress in blood cells could be the causes of these changes in the hematopoietic system. As a result, ATO affected bone marrow function (8). By attenuating the effects of the above variables, the administration of barely β-glucan and barley extracts decreased hepatotoxicity in arsenic-poisoned rats. The scavenging of free radicals by barley β-glucan and barley flour extracts could explain these results.
Previous studies have suggested that arsenic exposure changes haematological parameters, and these data support that hypothesis (1, 8). The absolute values of RBC count and haemoglobin serve as crucial indications of anaemia. ATO therapy can also cause immunological suppression in rats, due to alterations in lymphocyte and neutrophil populations (77). Monocytes are an important part of the immune system because they can also destroy invading pathogens through phagocytosis and antibody production. Monocyte levels were also reduced after treatment with ATO, presumably because of an association with inflammation (77).
The spleen's failure is the root cause of most illnesses. Modern medicine's immune function corresponds to the defence capacity (78). The immunological response in this investigation was compatible with the BW response, which is a clinical marker of illness. Furthermore, ATO has been demonstrated to damage the spleen and thymus glands, both of which are essential organs in the immune system. Rats' spleens and thymus glands lose weight.
In the case of oral administration treatment, the current study found that barely β-glucan extracts or barley flour extracts improved immunity in immune compromised rats. ATO administration reduced the weight of the spleen and thymus, indicating direct toxicity (79). These extracts, on the other hand, mitigated this decrease. Previous studies found that the barley and barely β-glucan groups had lower liver weights, kidney weights, and spleen weights (80, 12). These findings provide indirect evidence that barely β-glucan extracts or barley extracts boost immune responses in the body.
Histological alterations in the spleen are significantly linked to oxidative stress indicators. Histological investigations following arsenic trioxide treatment revealed lymphocytic necrosis and depletion with the appearance of tangible body macrophages, which were consistent with prior reports (79). Histopathological findings support the ability of barley β-glucan (80 and 160 mg/kg) and barley extracts (1.4 and 2.8 g/kg) to protect against spleen damage (with a corresponding decrease in MDA levels in the tissue). As a result, the findings show that β-glucan and barley extracts can be used as dietary supplements to protect humans and animals from arsenic toxicity. Barley can protect spleen tissue against steroid-induced structural alterations, according to Shehata et al. (81).
Barley β-glucan has been demonstrated to improve human health, particularly by lowering postprandial blood glucose and LDL cholesterol levels (82). Barley-glucans are widely available, easy to extract, and have high biological activity (83).