Amelioration of histopathological damage in mice with induced hyperlipidemia by terpenes and phytosterols extracted from Iraqi chickpea

Objective : To compare the amelioration effects of terpenes and phytosterols extracted from Iraqi chickpea on histopathological damage in mice fed on high-fat diet (HFD). Material and methods . Whole chickpea plants of the Fabaceae family were collected during the flowering period in the northern area of Erbil. The collected plants were cleaned, dried in a shaded area at room temperature, pulverized with mechanical mills, and weighed. Experiments were conducted from October 2021 to April 2022. The research involved 32 healthy albino male mice 2–3 months of age, weighing about 20–30 g. The animals were provided by the Higher Institute for Diagnosis of Infertility and Assisted Reproduction Techniques. One week prior to the onset of the experiment, the animals were acclimatized to standard environmental conditions; food and water were provided ad libitum. HFD (2% cholesterol and 1% peanut butter) was added to the standard diet for 28 days to induce hyperlipidemia. In all experimental groups, the body weight was measured weekly. The terpenes and phytosterol fraction extracted from Iraqi chickpea was administered for 28 days via an intragastric tube. Mice were kept fasting for 24 hours and blood samples were extracted via heart puncture. Standard diagnostic kits and an automated analyzer were used for estimation of serum total cholesterol (TC), triglycerides (TG), low-density lipoproteins (LDL), very lowdensity lipoproteins (VLDL), high-density lipoprotein, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total serum bilirubin (TSB) levels. The animals were sacrificed, then their liver and heart organs were removed for assay. Results. Histopathological examination of the hyperlipidemic mice group showed a marked and diffused cytoplasmic fatty infiltration of hepatic and cardiac cells. These effects were successfully ameliorated by administration of terpenes and phytosterols, although treatment with terpenes seemed to be more effective than that with phytosterols. In comparison with phytosterol treatment, terpene treatment in HFDinduced hyperlipidemic mice led to an improvement in lipid profiles, manifested in a highly significant decrease (p≤0.05) in the serum levels of TC (142.40±14.43 mg/dl), TG (98.89±8.71 mg/dl), LDL (79.43±15.14 mg/dl), and VLDL (19.78±1.74 mg/dl); with a highly significant (p≤0.05) decrease in the liver enzymatic activities of (ALT (19.19±1.36 U/l), AST (15.98±1.3 U/l), and ALP (20.99±4.43 U/l)) and TSB levels (1.60±0.12 mg/dl) (highly significantly differences with p≤0.05). Terpenes also led to a statistically significant improvement in the level of tissue malondialdehyde and glutathione in hyperlipidemic mice (p≤0.05). Conclusion . In comparison with phytosterols, terpenes exhibit a highly significant ameliorating effect in HFD-induced hyperlipidemia, improving the state of liver and heart tissue, lipid profile, liver function enzymes, and oxidative stress parameters. Further research should assess the efficacy of terpenes and phytosterols and to elucidate the mechanism of their anti-hyperlipidemic action.