Tong Jing, Yao Xincheng, Zeng Hong, Zhou Gao, Chen Yuxin, Ma Bingxin, Wang Youwei
Institute of TCM & Natural Products, School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, PR China.
Institute of TCM & Natural Products, School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, PR China; College of Pharmacy, Shihezi University, Xinjiang Uyghur Autonomous Region, Shihezi 832000, PR China.
J Ethnopharmacol. 2015 Nov 4;174:355-63. doi: 10.1016/j.jep.2015.08.045. Epub 2015 Aug 29.
Cichorium glandulosum Boiss. et Huet was used historically in Uyghur folk medicine. Its roots, seeds, and aerial parts are extensively used by Uyghur residents in Xinjiang to eliminate savda typhoid, dredge and cure obstructive jaundice variety liver disorders.
To evaluate the hepatoprotective activity of total flavonoids (TFs) obtained from C. glandulosum seeds against carbon tetrachloride (CCl4)-induced liver damage in vitro and in vivo. To investigate the mechanisms of hepatoprotective effects for TFs.
The dried seeds of C. glandulosum were extracted with 70% aqueous ethanol, and the extract was chromatographed with D101 macroporous resin. In vitro the antioxidant capacity against lipid peroxidation (LPO) was evaluated using ferrothiocyanate, thiobarbituric acid, β-carotene bleaching, and LPO inhibition assay. The cytotoxicity and hepatoprotective activity of TFs were evaluated in human liver hepatoma cells (HepG2). MTT assay, hepatic injury markers aspartate aminotransferase (AST), alanine aminotransaminase (ALT), lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA), and glutathione (GSH) were performed. In vivo the hepatoprotective activity of TFs against CCl4-induced acute liver injury was evaluated in rats. A series of biochemical and antioxidant parameter levels were measured in liver homogenate. The suppressive effect on pancreatic lipase activity was determined.
Results indicated that TFs showed antioxidant capacity against lipid peroxidation (LPO). Administrating CCl4 (1%, v/v) caused a significant decrease in HepG2 viability. Treatment with TFs at doses (62.5, 125, and 250 μg/ml) could significantly ameliorate the cytotoxicity and decline the levels of AST, ALT, and LDH induced by CCl4. The markers including MDA and GSH, which were close to oxidative damage, were restored. Oral treatment with TFs in vivo at doses of 100, 200, and 400 mg/kg significantly reduced the levels of AST, ALT, alkaline phosphatase (ALP), total bilirubin (TB), and thiobarbituric acid reactive substances (TBARs) in the serum compared with CCl4-induced acute liver injury in rats. TFs showed dose-dependent suppressive effects on pancreatic lipase activity, and the IC50 was 1.318 ± 0.164 mg/ml.
TFs from C. glandulosum seeds demonstrated significant hepatoprotection against CCl4-induced hepatotoxicity. TFs exhibited significant suppression of LPO and pancreatic lipase capacity, which may be the mechanisms of hepatoprotective effects against CCl4. Our results contribute towards validation of the traditional use of C. glandulosum seeds in the treatment of liver disorders.
腺毛菊苣（Cichorium glandulosum Boiss. et Huet）在维吾尔族民间医学中有着悠久的使用历史。新疆的维吾尔族居民广泛使用其根、种子和地上部分来消除“沙吾达”伤寒，疏通并治疗阻塞性黄疸等多种肝脏疾病。
评估从腺毛菊苣种子中提取的总黄酮（TFs）在体外和体内对四氯化碳（CCl4）诱导的肝损伤的保肝活性。探究总黄酮保肝作用的机制。
腺毛菊苣干燥种子用70%乙醇水溶液提取，提取物用D101大孔树脂进行柱色谱分离。体外使用硫氰酸铁、硫代巴比妥酸、β-胡萝卜素漂白和脂质过氧化抑制试验评估对脂质过氧化（LPO）的抗氧化能力。在人肝癌细胞（HepG2）中评估总黄酮的细胞毒性和保肝活性。进行MTT试验、检测肝损伤标志物天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）、乳酸脱氢酶（LDH）泄漏、丙二醛（MDA）和谷胱甘肽（GSH）。在体内评估总黄酮对CCl4诱导的大鼠急性肝损伤的保肝活性。测定肝匀浆中一系列生化和抗氧化参数水平。测定对胰脂肪酶活性的抑制作用。
结果表明总黄酮对脂质过氧化（LPO）具有抗氧化能力。给予CCl4（1%，v/v）导致HepG2细胞活力显著下降。用总黄酮（62.5、125和250μg/ml）处理可显著改善细胞毒性，并降低CCl4诱导的AST、ALT和LDH水平。接近氧化损伤的MDA和GSH等标志物得以恢复。与CCl4诱导的大鼠急性肝损伤相比，总黄酮以100、200和400mg/kg的剂量口服给药可显著降低血清中AST、ALT、碱性磷酸酶（ALP）、总胆红素（TB）和硫代巴比妥酸反应物质（TBARs）的水平。总黄酮对胰脂肪酶活性表现出剂量依赖性抑制作用，IC50为1.318±0.164mg/ml。
腺毛菊苣种子中的总黄酮对CCl4诱导的肝毒性具有显著的保肝作用。总黄酮对LPO和胰脂肪酶活性表现出显著抑制作用，这可能是其对CCl4保肝作用的机制。我们的结果有助于验证腺毛菊苣种子治疗肝脏疾病的传统用途。
