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目的观察小檗碱对非酒精性脂肪性肝病(NAFLD)大鼠肝组织p38MAPK-STAT3信号通路相关基因及蛋白表达的调控作用,探讨小檗碱防治NAFLD的作用机制。方法取SPF级雄性SD大鼠24只,适应性喂养1周后,随机分为正常组、模型组和小檗碱组(100 mg·kg~(-1)),每组8只。除正常组给予基础饲料外,其余两组均采用高脂饮食喂养,建立大鼠NAFLD模型,在施以造模因素的同时,小檗碱组灌胃给药,8周后对肝组织行HE染色和油红O染色。全自动生化仪检测血清总胆固醇(TC)、甘油三脂(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、谷丙转氨酶(ALT)、谷草转氨酶(AST);ELISA法检测血清及肝组织超氧化物歧化酶(SOD)、丙二醛(MDA)含量;实时荧光定量RT-PCR法检测肝组织p38MAPK、STAT3 m RNA表达量;Western Blotting法检测肝组织蛋白p38MAPK、STAT3及p-p38MAPK、p-STAT3表达水平。结果 8周高脂饮食成功诱导建立了NAFLD实验动物模型,HE染色和油红O染色证实大鼠模型肝组织脂质蓄积严重。与正常组比较,模型组大鼠血清TG、TC含量明显升高(P<0.05),肝组织及血清SOD活力明显降低(P<0.01),MDA水平明显升高(P<0.01),p38MAPK、STAT3 m RNA及p38MAPK、p-p38MAPK、STAT3、p-STAT3蛋白的表达水平均显著升高(P<0.05,P<0.01);与模型组比较,小檗碱组大鼠血清TG、TC含量明显降低(P<0.05),肝组织及血清SOD活力明显升高(P<0.05,P<0.01),MDA水平明显降低(P<0.05,P<0.01),p38MAPK、STAT3 m RNA及p-p38MAPK、p-STAT3蛋白表达水平明显下调(P<0.05)。结论小檗碱可以改善NAFLD模型大鼠肝脏脂肪变性,降低其血清及肝组织氧化应激因子水平,其机制可能是通过激活肝脏p38MAPK-STAT3信号通路相关基因及其蛋白磷酸化水平,改善氧化应激而发挥作用。
Objective To observe the regulatory effect of berberine on the expression of p38MAPK-STAT3 signal pathway and its protein in non-alcoholic fatty liver disease (NAFLD) rats and to explore the mechanism of berberine in preventing and treating NAFLD. Methods Twenty-four SPF male Sprague Dawley rats were randomly divided into normal group, model group and berberine group (100 mg · kg -1) for one week after adaptive feeding. In addition to the normal group given the basic diet, the other two groups were fed with a high-fat diet to establish NAFLD model rats, while modeling factors, berberine group gavage, 8 weeks after the liver tissue HE Dyeing and Oil Red O staining. The levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST). The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and liver tissue were detected by ELISA. The expression of p38MAPK and STAT3 mRNA in liver tissues were detected by real-time fluorescent quantitative RT-PCR. The expression of p38MAPK, STAT3, p-p38MAPK and p-STAT3 in liver tissue were detected. Results Eight weeks of high-fat diet induced NAFLD experimental animal model was established, HE staining and oil red O staining confirmed rat model liver lipid accumulation was serious. Compared with the normal group, the levels of serum TG and TC in model group were significantly increased (P <0.05), the activity of SOD in liver tissue and serum were significantly decreased (P <0.01), the level of MDA was significantly increased (P <0.01) The levels of STAT3 m RNA, p38MAPK, p-p38MAPK, STAT3 and p-STAT3 were significantly increased (P <0.05, P <0.01). Compared with the model group, the contents of TG and TC in the serum of berberine group (P <0.05, P <0.01, P <0.05, P <0.01). The levels of p38MAPK, STAT3mRNA and p-p38MAPK in liver tissue and serum were significantly increased p-STAT3 protein expression was significantly down-regulated (P <0.05). Conclusions Berberine can improve hepatic steatosis and decrease the levels of oxidative stress factors in serum and liver of NAFLD rats. The mechanism may be related to the activation of oxidative stress by activating the gene and protein phosphorylation of p38MAPK-STAT3 signal pathway Excitement and play a role.