1 Plant Archives Vol. 20, Supplement 1, 2020 pp. 3157-3164 e-ISSN:2581-6063 (online), ISSN:0972-5210 ANISOTES TRISULCUS PREVENTS HYPERCHOLESTEROLEMIA, OXIDATIVE STRESS AND INFLAMMATION AND MODULATES FATTY ACID SYNTHASE IN HIGH CHOLESTEROL DIET-FED RATS Mousa O. Germoush Biology Department, College of Science, Jouf University, Sakaka-2014, Saudi Arabia E-mail: mousagermoush@gmail.com & mogermoush@ju.edu.sa Abstract Anisotes trisulcus is a stiff erect shrub with antioxidant and anti-inflammatory activities. This study investigated the protective effect of A. trisulcus extract against oxidative stress, inflammation and hypercholesterolemia in high cholesterol diet (HCD)- fed rats. Rats received HCD and A. trisulcus extract (100 and 200 mg/kg) for 10 weeks and samples were collected for analysis. Serum cholesterol, triglycerides, LDL-cholesterol and vLDL-cholesterol along with cardiovascular risk indices were significantly increased in HCD-fed rats. HDL-cholesterol was decreased, and pro-inflammatory cytokines and ALT, AST and ALP were increased in serum of HCD-fed rats. Treatment with A. trisulcus extract ameliorated these metabolic alterations and decreased hepatic lipid peroxidation levels. In addition, A. trisulcus extract enhanced hepatic GSH and antioxidant enzymes in HCD-fed rats. Furthermore, A. trisulcus extract decreased the expression of hepatic fatty acid synthase (FAS) and stimulated a trend increase in LDL receptor in HCD-fed rats. In conclusion, this study showed the anti-hypercholesterolemic, antioxidant and anti-inflammatory activities of A. trisulcus in HCD-fed rats. A. trisulcus ameliorated serum lipids, cardiovascular risk indices, liver function markers and pro-inflammatory cytokines, and hepatic oxidative stress and FAS expression. Keywords : Almodh; LDLR; FAS; Cholesterol; Oxidative stress; Inflammation. Introduction Hyperlipidemia/dyslipidemia has been demonstrated as a casual risk factor for atherosclerotic cardiovascular disease (Nordestgaard, 2016). It is a disorder of lipid metabolism and manifested as increased levels of low density lipoprotein (LDL) cholesterol and triacylglycerides (Nordestgaard, 2016). Hyperlipidemia may arise through lifestyle factors such as excessive consumption of foods rich in cholesterol and saturated fats, but is also highly heritable (Ma et al., 2012, Yuan et al., 2007). Hypercholesterolemia is a type of hyperlipidemias that elicits atherosclerosis, chronic inflammation and accumulation of hepatic lipids (steatosis) and hence reduce the ability of the liver to lower circulating lipids (Lee et al., 2017). The role of hypercholesterolemia in eliciting oxidative stress has been demonstrated in different studies (Al-Rejaie et al., 2013; Lee et al., 2017; Olorunnisola et al., 2012). The accumulation of cholesterol in different cells, such as hepatocytes and endothelial cells has been associated with diminished antioxidant defenses and excess production of reactive oxygen species (ROS) (Anila & Vijayalakshmi, 2003; Forstermann, 2008). Excess ROS accumulation can induce inflammation, cell death and metabolic alterations (Jones, 2006; Seifried et al., 2007). Hence, agents that can counteract hypercholesterolemia and other hyperlipidemias and exert antioxidant and anti- inflammatory activities can protect against the metabolic alterations induced by high levels of lipids. Medicinal plants are rich sources of bioactive compounds that can exert anti-hyperlipidemic effects accompanied with attenuation of oxidative stress and inflammation (Aladaileh et al., 2019b). Anisotes trisulcus (family Acanthaceae) is a stiff erect shrub and commonly known as Almodh in Saudi Arabia (El-Shanawany et al., 2011). A. trisulcus has been reported to exert multiple activities, including ant-hypertension, anti-bacterial, local anesthetic and hepatoprotective. In addition, this species is used to limit tobacco consumption, suppress appetite and traditionally in the treatment of jaundice, hepatitis, gallstones and other disorders related to the liver (Al-Rehaily et al., 2011; Al-Rehaily et al., 2002; Ali et al., 2001; El-Shanawany et al., 2011). A. trisulcus has been demonstrated to be a rich source of phenolic compounds, including vanillic acid, veratric acid, -sitosterol, stigmasterol, -amyrin and veratric acid, and its methanolic extract exhibited antioxidant and anti-inflammatory activities (El-Shanawany et al., 2014). However, the beneficial effects of A. trisulcus on hypercholesterolemia have not been previously investigated. Therefore, this study evaluated the anti-hypercholesterolemic effect of A. trisulcus and its protective antioxidant and anti- inflammatory activities in high cholesterol diet (HCD)-fed rats. Additionally, the ability of A. trisulcus to modulate hepatic fatty acid synthase (FAS) and LDL receptor (LDLR) expression in HCD-fed rats was studied. Materials and Methods A. trisulcus leaf collection and extraction The leaves of A. trisulcus were collected from the northern are of Saudi Arabis (Sakaka) and were identified and authenticated by an expert taxonomist from the Botany Department, College of Science, Jouf University, KSA. The leaves were washed under running tap water, dried in the shade and then mashed into a powder size and macerated with methanol (80%; v/v) at 4°C for 72 h. The macerated product was filtered, and the obtained supernatant was concentrated using a rotary evaporator to a semi-dry state and then dissolved in distilled water. Animals and experimental design Thirty adult male Wistar rats weighing 140-160 g were housed under standard laboratory conditions (12 h light/dark