摘要
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： Observation of the preventive effects of TCM therapies on NASH and the AMPK mRNA expression activated by AMP in rats＇ livers, to discuss its possible mechanism to prevent and treat NASH. Methods： 70 rats were randomly divided into 7 groups, including the normal group, model group, the group of soothing the liver, the group of reducing phlegm, the group of tonifying kidney, the group of invigorating the circulation of blood and the metformin group, with 10 rats in each group. The rats in the normal group were fed with the standard diet, and the rats in the other groups were fed with fat- rich diet. Meanwhile, the rats in every treatment group were respectively fed with 6g.kg-1.d-1 of Chaihu Shugan San, 14g.kg-1.d-1 Erchen Tang combined with Siling San, 17g.kg-1.d-1 of Jinkui Shenqi Wan, 14g.kg-1.d-1 of Gexia Zhuyu Tang and 60mg.kg -1.d-1 of metformin by daily garage. The rats in the normal and model groups were continuously fed with distilled water in 12 weeks. The serum lipid, liver lipid and serum enzymology were detected with biochemistry method. The liver pathological change was evaluated under light microscope after HE staining, serum FFA was examined by ELISA and the expression of liver AMPKαmRNA and ACC2mRNA were detected by Real Time-PCR. Results： The NAS of the rats in the model group were remarkably higher than that in the normal group （P〈0.01）, the disorder of serum lipid and amounts of hepatic TG, CHOL and serum ALT, AST of the rats in the model group were remarkably higher than those in the normal group （P〈0.01）, the AMPK mRNA expression of liver in the model rats was lower than that of the normal and ACC2mRNA expression was higher. After the treatment with Chaihu Shugan San, Erchen Tang combined with Siling San, Jinkui Shenqi Wan, Gexia Zhuyu Tang and metformin, the NAS of the rats in the treatment groups were remarkably higher than that in the normal group （P〈0.05）. The contents of serum CHOL, FFA, ALT and AST and liver TG, CHOL of the rats in the model group were remarkably higher than that in the normal group （P〈0.01, P〈0.05）, simultaneously the liver AMPKctmRNA expression in the groups of the soothing the liver, the reducing phlegm, and invigorating the circulation of blood was obviously higher than the model group （P〈0.01, P〈0.05）, and the ACC2mRNA expression was lower. But the liver expression of AMPKamRNA and ACC2mRNA in the the group of tonifying kidney showed no significant difference with the model （P〉0.05）. Conclusions： The therapies of Soothing the liver and regulating vital energy, tonifying spleen and reducing phlegm, and promoting blood circulation and removing blood stasis could correct the disorder of lipid metabolism in rats with NASH, and relieve the degree of NASH by means of reducing the hepatic inflammation in rats with varying degrees. Its mechanism maybe associated with activating AMPKα, down-regulating the expression of target gene ACC2mRNA. Jinkui Shenqi Wan probably regulate the lipid disorder of rats with other ways of regulation and control.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2014年第1期277-281,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
浙江省自然科学基金资助项目(No.Y2100170)~~
