溪黄草水提物对四氯化碳诱导大鼠肝纤维化的保护作用及机制研究
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篇名: 溪黄草水提物对四氯化碳诱导大鼠肝纤维化的保护作用及机制研究
TITLE:
摘要: 目的:研究溪黄草水提物(RWE)对四氯化碳(CCl4)诱导大鼠肝纤维化(HF)的保护作用及其机制。方法:将60只雄性SD大鼠随机分为正常组、模型组、秋水仙碱组(0.12 mg/kg)和RWE低、中、高剂量组(4、8、16 g/kg,以生药量计),每组10只。除正常组大鼠腹腔注射橄榄油外,其余各组大鼠均腹腔注射40% CCl4橄榄油溶液以复制HF模型。自造模第1天起,各给药组大鼠均灌胃相应药物(10 mL/kg),正常组和模型组大鼠均灌胃等体积水,每天1次,连续6周。给药结束后,采用生化法或酶联免疫吸附测定法检测大鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)的含量以及肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-1β的含量或活性;采用苏木精-伊红染色观察大鼠肝组织病理学变化;采用蛋白质印迹法检测大鼠肝组织中α-平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGF-β1)蛋白表达情况。结果:与正常组比较,模型组大鼠血清中ALT、AST、HA、LN、PCⅢ、Ⅳ-C的含量以及肝组织中MDA、TNF-α、IL-6、IL-1β的含量均显著升高(P<0.01),肝组织中SOD、GSH-Px的活性均显著降低(P<0.01);肝组织纤维化明显;肝组织中α-SMA、TGF-β1蛋白的相对表达量均显著升高(P<0.01)。与模型组比较,秋水仙碱组和RWE各剂量组大鼠血清中ALT、AST、HA、LN、PCⅢ、Ⅳ-C的含量以及肝组织中MDA、TNF-α、IL-6的含量,秋水仙碱组和RWE中、高剂量组大鼠肝组织中IL-1β的含量均显著降低(P<0.05或P<0.01),秋水仙碱组和RWE各剂量组大鼠肝组织中SOD、GSH-Px的活性均显著增强(P<0.05或P<0.01);肝组织纤维化程度明显减轻;肝组织中α-SMA、TGF-β1蛋白的相对表达量均显著降低(P<0.01)。结论:RWE对CCl4诱导的HF模型大鼠具有一定的保护作用,其作用机制可能与调节脂质代谢、减轻肝脂质过氧化损伤、抗氧化应激反应、抑制炎症因子释放及TGF-β1蛋白表达等有关。
ABSTRACT: OBJECTIVE: To study the protective effects and the mechanism of Rabdosia serra water extract (RWE) on hepatic fibrosis (HF) induced by carbon tetrachloride (CCl4) in rats. METHODS: Sixty male SD rats were randomly divided into normal group, model group, colchicine group (0.12 mg/kg), and RWE low-dose, medium-dose and high-dose groups (4, 8, 16 g/kg, by crude drug), with 10 rats in each group. Except for intraperitoneal injection of olive oil for normal group, other groups were given 40% CCl4 olive oil solution intraperitoneally to induce HF model. Since the first day of modeling, each treatment group was given relevant medicine (10 mL/kg) intragastrically, while normal group and model group were given constant volume of water intragastrically, once a day, for consecutive 6 weeks. After medication, biochemical process or ELISA were used to determine the contents of ALT, AST, HA, LN, PCⅢ and Ⅳ-C in serum, the activities or contents of SOD, GSH-Px, MDA, TNF-α, IL-6 and IL-1β in liver tissue. Pathological changes of liver tissue in rats were observed by HE staining. The expression of α-SMA and TGF-β1 in liver tissue were detected by Western blot. RESULTS: Compared with normal group, the contents of ALT, AST, LN, HA, PCⅢ and Ⅳ-C in serum, the contents of MDA, TNF-α, IL-6 and IL-1β in liver tissue were all increased significantly in model group (P<0.01); the activities of SOD and GSH-Px in liver tissue were decreased significantly (P<0.01). Liver fibrosis was obvious, and the relative expression of α-SMA and TGF-β1 were increased significantly (P<0.01). Compared with model group, the contents of ALT, AST, HA, LN, PCⅢ and Ⅳ-C in serum as well as the contents of MDA, TNF-α and IL-6 in liver tissue in colchicines group and RWE groups, the contents of IL-1β in liver tissue of rats in colchicines group, RWE medium-dose and high-dose groups were all decreased significantly (P<0.05 or P<0.01). The activities of SOD and GSH-Px in liver tissue of rats were increased significantly in colchicines group and RWE groups (P<0.05 or P<0.01). The fibrosis degree of liver tissue was significantly reduced, while the relative expression of α-SMA and TGF-β1 decreased significantly (P<0.01). CONCLUSIONS: RWE can protect CCl4-induced HF model rats, the mechanism of which may be associated with regulating lipid metabolism, relieving liver lipid peroxidation injury and anti-oxidative stress response, inhibiting the release of inflammatory factors and the expression of TGF-β1.
期刊: 2018年第29卷第20期
作者: 许琼梅,李跃龙,曹后康,陈春,王刚,朱依谆,张可锋
AUTHORS: XU Qiongmei,LI Yuelong,CAO Houkang,CHEN Chun,WANG Gang,ZHU Yizhun,ZHANG Kefeng
关键字: 溪黄草;水提物;肝纤维化;四氯化碳;氧化应激;炎症因子;转化生长因子β1;大鼠
KEYWORDS: Rabdosia serra; Water extract; Hepatic fibrosis; Carbon tetrachloride; Oxidative stress; Inflammatory factor; TGF-β1; Rats
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