تقييم الدور الوقائي للمستخلص النانوي والكحولي لفاكهة التين الشوكي Opuntia ficus-indica في بعض المعايير النسجية والفسلجية-المناعية في ذكور الجرذان البيض المعاملة بمادة Thioacetamide

Evaluation of the Protective Role of the Nano and Alcoholic Extracts of Opuntia ficus-indica Fruit on some Histological and Physiological–Immunological Parameters in Male Albino Rats Treated with Thioacetamide

The present study aimed to evaluate the protective role of the alcoholic extract and the biosynthesized zinc oxide nanoparticles derived from the peel and pulp of the prickly pear fruit Opuntia ficus-indica against the physiological, immunological, and histopathological alterations induced in the liver and kidneys by thioacetamide (TAA) in male albino rats.
The study was conducted at the Postgraduate Laboratory, College of Education for Pure Sciences, University of Kerbala, and the Animal House, College of Pharmacy, University of Kerbala, during the period from July 15, 2024, to December 15, 2024. The study included the use of 72 male albino rats, aged 10–12 weeks and weighing 200–250 g. The alcoholic and nano extracts of the peel and pulp of Opuntia ficus-indica were prepared and the research comprised two experiments, The first experiment aimed to determine the median effective dose (ED50) of the nano extract of the peel and pulp of Opuntia ficus-indica by studying the dose–response curve. In this experiment, 36 male albino rats were used and randomly divided into six equal groups (6 animals per group). The animals were orally administered five ascending doses of the nano extract at 20, 40, 60, 80, and 100 mg/kg, in addition to the control group, which was administered ordinary water, for a period of 30 days. At the end of the experiment, blood samples were collected to assess the following parameters: malondialdehyde (MDA) levels, reduced glutathione (GSH), total cholesterol (TC), and high-density lipoprotein (HDL). The median effective dose (ED50) of the nano extract was found to be 50 mg/kg for the peel and 54 mg/kg for the pulp of Opuntia ficus-indica.
The second experiment was designed to investigate the protective effects of the alcoholic and nano extracts of the peel and pulp of Opuntia ficus-indica against the hepatotoxic and nephrotoxic effects induced by thioacetamide (TAA). A total of 36 male albino rats were used in this experiment, randomly divided into six equal groups (6 animals per group). The experiment lasted for 14 weeks. Group 1 (G1): administered ordinary water and considered as the negative control group. Group 2 (G2): intraperitoneally injected with TAA at a dose of 200 mg/kg, dissolved in distilled water, twice per week. Group 3 (G3): orally administered the alcoholic extract of the pulp of Opuntia ficus-indica at a dose of 100 mg/kg, followed by TAA injection. Group 4 (G4): orally administered the alcoholic extract of the peel of Opuntia ficus-indica at a dose of 100 mg/kg, followed by TAA injection. Group 5 (G5): orally administered the nano extract of the pulp of Opuntia ficus-indica at a dose of 54 mg/kg, followed by TAA injection. Group 6 (G6): orally administered the nano extract of the peel of Opuntia ficus-indica at a dose of 50 mg/kg, followed by TAA injection.
At the end of the experiment, and following an 8-hour fasting period, blood samples were collected from the animals for the analysis of physiological, immunological, and molecular markers. The assessments included the measurement of: ALT, AST, ALP, T-BIL, Albumin (Alb), Creatinine (Cr), Urea (Ur), GSH, MDA, SOD, COX, TNF-α, IL-1β, IL-6, CEA, BCL-2, Caspase-3, TGF-β, and MCP-1. Subsequently, the liver and kidneys were excised and preserved in formalin for histopathological examination.
The results of this study revealed that intraperitoneal injection of TAA in the positive control group (G2) caused a significant increase (P ≤ 0.05) in the levels of ALP, AST, ALT, T-BIL, Creatinine, Urea, MDA, TNF-α, CEA, IL-6, IL-1β, BCL-2, Caspase-3, TGF-β, and MCP-1, accompanied by a significant decrease (P ≤ 0.05) in the levels of GSH, SOD, COX, and Albumin, when compared with the negative control group (G1). In contrast, male rats in groups G3 and G4, which were administered the alcoholic extracts of the pulp and peel of Opuntia ficus-indica, respectively, as well as groups G5 and G6, which were administered the nano extracts of the pulp and peel of Opuntia ficus-indica, respectively, exhibited a significant increase (P ≤ 0.05) in the levels of SOD, GSH, COX, and Albumin. This was accompanied by a significant decrease (P ≤ 0.05) in the levels of ALT, AST, ALP, MCP-1, IL-1β, IL-6, TNF-α, MDA, Creatinine, Urea, T-BIL, CEA, Caspase-3, and TGF-β compared with group G2.
Histological sections of the liver tissue from the groups of rats treated with TAA at a concentration of 200 mg/kg revealed severe pathological alterations. These included congestion and dilation of the central vein, marked infiltration of inflammatory cells, hepatocellular necrosis, disorganization of hepatic cords, evident degenerative changes, sinusoidal congestion, as well as the presence of tumorous nodules and fibrosis in some samples, compared with group G1. In contrast, the groups administered the alcoholic extracts of the pulp and peel of Opuntia ficus-indica (G3, G4) showed a clear improvement characterized by better organization of hepatic cords, integrity of hepatocytes and their nuclei, with mild congestion of the central vein and slight inflammatory cell infiltration. Meanwhile, the groups treated with the nano extracts of the pulp and peel of Opuntia ficus-indica (G5, G6) displayed liver tissue architecture that was closer to the normal state, with well-defined central veins, normal appearance of hepatocytes, and clearly visible nuclei, in comparison with group G2.
Histological sections of the kidney tissue from the groups of rats treated with TAA at a concentration of 200 mg/kg exhibited severe pathological alterations. These included glomerular atrophy, dilation of Bowman’s space, destruction of the tubular walls, and detachment of the lining epithelium. Marked degenerative changes were also observed in the renal tubules, along with vascular congestion, necrosis of renal cells, and inflammatory cell infiltration within the interstitial tissue, compared with group G1. In contrast, the groups administered the alcoholic extracts of Opuntia ficus-indica (pulp and peel) (G3, G4) showed the presence of glomeruli and Bowman’s capsule, with vascular congestion and some hemorrhagic manifestations, while the renal tubules largely preserved their normal architecture, particularly in the peel-treated group. Meanwhile, the groups treated with the nano extracts of Opuntia ficus-indica (pulp and peel) (G5, G6) displayed kidney tissue architecture that was closer to the normal state, with intact glomerular structures, well-defined Bowman’s capsule and space, and nearly normal architecture of the renal tubules, without evident damage when compared with group G2.
The present study concluded that the extracts of Opuntia ficus-indica, whether alcoholic or nano, exerted a clear protective effect against TAA-induced hepatotoxicity and nephrotoxicity in male albino rats. This protective role was demonstrated through the improvement of liver and kidney functions by reducing biomarkers of tissue injury, decreasing oxidative stress and inflammatory indicators, enhancing natural antioxidant defenses, and improving the histological architecture of the liver and kidneys. Moreover, the nano extracts—particularly the nano extract of the prickly pear peel—showed superior efficacy in providing broader histological and functional protection compared with the alcoholic extracts. These findings suggest the potential applicability of such extracts in preventing the toxic effects of harmful agents on the liver and kidneys and in reducing the risk of hepatocellular carcinoma or tumor development.