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目的:观察不同中医治法方药对非酒精性脂肪性肝炎(NASH)的防治作用及对大鼠肝脏腺苷酸激活蛋白激酶(AMPK)αmRNA表达的影响,探讨其防治NASH的可能机制。方法:大鼠随机分为正常组、模型组、疏肝组、化痰组、补肾组、活血组及二甲双胍组7组,每组10只。正常组予标准饲料喂养,其他各组每天予高脂饲料喂养,同时疏肝组、健脾组、补肾组、活血组及二甲双胍组分别予6g·kg-1·d-1的柴胡疏肝散、14g·kg-1·d-1的二陈汤合四苓散、17g·kg-1·d-1的金匮肾气丸、14g·kg-1·d-1的膈下逐瘀汤以及60mg·kg-1·d-1的二甲双胍灌胃,正常组和模型组每天以等容量的蒸馏水灌胃,均连续12周。生化法检测血脂、肝脂、血清酶学指标;HE染色光镜下观察肝组织病理学变化;ELISA法检测大鼠血清游离脂肪酸(FFA)水平;RT-PCR法检测肝组织AMPKαmRNA、ACC2 mRNA的表达。结果:模型组大鼠肝组织非酒精性脂肪性肝病活动度积分(NAS)较正常组明显增高(P<0.01),血脂紊乱,肝组织TG、总胆固醇(CHOL)含量及血清丙氨酸转氨酶(ALT)、天门冬酸转氨酶(AST)水平较正常组显著升高(P<0.01);肝组织AMPKαmRNA表达较正常组显著降低(P<0.01);ACC2mRNA表达较正常组明显增高;各给药组肝组织NAS均较模型组有不同程度下降(P<0.05),血清CHOL、FFA、ALT、AST及肝组织TG、CHOL水平较模型组不同程度降低(P<0.01,P<0.05),同时疏肝组、健脾组、活血组大鼠肝组织AMPKαmRNA表达较模型明显增强(P<0.01,P<0.05),ACC2 mRNA表达较模型明显减少,但补肾组肝组织AMPKαmRNA和ACC2 mRNA表达则与模型组差异无统计学意义。结论:疏肝理气、健脾化痰、活血化瘀等不同中医治法方药能不同程度地纠正NASH大鼠脂质代谢紊乱,减轻肝组织炎症损伤,防止NASH的进一步发展,其机制可能通过激活AMPKα,下调其靶基因的表达实现;温补脾肾代表金匮肾气丸调节则可能通过其他的调控途径调节NASH大鼠脂质代谢紊乱防止NASH的进展。
Objective: To observe the effects of different traditional Chinese medicine prescriptions on the prevention and treatment of non-alcoholic steatohepatitis (NASH) and the expression of AMPK mRNA in rat liver and to explore the possible mechanism of prevention and treatment of NASH. Methods: The rats were randomly divided into normal group, model group, Shugan group, Huatan group, Bushen group, Huoxue group and metformin group, with 7 in each group. The normal group was fed with standard feed, while the other groups were fed with high-fat diet every day. At the same time, Shugan group, Jianpi group, Bushen group, Huoxue group and metformin group were administered with 6g · kg-1 · d- , 14g · kg-1 · d-1 Erchen Tang combined with Siling Powder, 17g · kg-1 · d-1 Jinguishenqi Pills, 14g · kg-1 · d-1 Gexia Zhuyu Decoction and 60 mg · kg-1 · d-1 metformin was orally administered to the normal group and model group by intragastric administration of distilled water of equal capacity for 12 consecutive weeks. Serum lipids, hepatic lipids and serum enzymological indexes were detected by biochemical methods; pathological changes of liver tissues were observed by HE staining; Free fatty acids (FFA) levels were measured by ELISA; AMPKα mRNA and ACC2 mRNA expression. Results: Compared with the normal group, the activity integral (NAS) of nonalcoholic fatty liver disease in the model group was significantly higher than that in the normal group (P <0.01), and the levels of lipid, TG and CHOL in the liver tissue, serum alanine aminotransferase (P <0.01). The expression of AMPKαmRNA in hepatic tissue was significantly lower than that in normal group (P <0.01), and the expression of ACC2mRNA in hepatic tissue was significantly higher than that in normal group Compared with the model group, NAS decreased significantly (P <0.05), serum CHOL, FFA, ALT, AST and liver tissue TG, CHOL levels were lower than the model group (P <0.01, P <0.05) Compared with model, the expression of AMPKαmRNA in liver tissue of Shugan group, Jianpi group and Huoxue group were significantly increased (P <0.01, P <0.05), while the expression of ACC2 mRNA in model group was lower than that of model group There was no significant difference in model group. Conclusion: Shuganqiqi, spleen-resolving phlegm, promoting blood circulation and removing blood stasis and other traditional Chinese medicine prescriptions can correct the disorder of lipid metabolism in NASH rats to some degree, reduce the inflammatory injury of liver tissue and prevent the further development of NASH. The mechanism may be through activating AMPKα, and down-regulate the expression of its target gene. The regulation of WBP on behalf of Jingui Shenqi Pill may regulate the lipid metabolism disorders in NASH rats and prevent the progression of NASH through other regulatory pathways.