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目的:观察疏肝健脾方药对非酒精性脂肪性肝病(NASH)大鼠肝组织TLR4 mRNA及蛋白表达的影响,探讨疏肝健脾方药抗大鼠NASH的作用机制。方法:SPF级雄性SD大鼠72只,随机分为正常对照组、模型组、疏肝高剂量组(9.6 g/kg柴胡疏肝散)、疏肝低剂量组(3.2 g/kg柴胡疏肝散)、健脾高剂量组(30 g/kg参苓白术散)、健脾低剂量组(10 g/kg参苓白术散)、合方高剂量组(39.6 g/kg柴胡疏肝散合参苓白术散)、合方低剂量组(13.2 g/kg柴胡疏肝散合参苓白术散),每组9只。采用高脂饲料(10 mL/kg)喂养建立NASH大鼠模型,16 w后各组大鼠以3%戊巴比妥钠腹腔麻醉,腹主动脉采血,全自动生化仪检测血脂、肝脂和肝功能;HE和油红O染色观察肝组织病理变化;Real time Q-PCR检测肝组织TLR4 mRNA表达水平;Western blot检测肝组织TLR4蛋白表达水平。结果:与正常对照组比较,模型组大鼠肝细胞脂肪变性明显,肝细胞肿大,小叶内、汇管区炎细胞浸润,血清TC、LDL-C,肝组织TC、TG、肝组织TLR4 mRNA和蛋白表达均显著升高(P<0.01);与模型组比较,各用药组大鼠血脂、肝脂、肝组织TLR4 mRNA和蛋白表达均有不同程度降低(P<0.05或P<0.01),以高剂量作用最为显著。结论:疏肝健脾方药可能是通过下调TLR4 mRNA和蛋白表达水平发挥抗NASH的作用,TLR4可能是疏肝健脾方药抗NASH的有效靶点之一。疏肝健脾方药防治NASH可能存在一定的量效依赖关系。
Objective: To observe the effects of Shuganjianpi Decoction on TLR4 mRNA and protein expression in the liver of non-alcoholic fatty liver disease (NASH) rats and to explore the mechanism of action of Shuganjianpi Decoction against NASH in rats. Methods: Seventy two Sprague-Dawley male Sprague-Dawley rats were randomly divided into three groups: normal control group, model group, Shugan high dose group (9.6g / kg Chaihu Shugan San), Shugan low dose group (3.2g / kg Bupleurum Shugunsan), high-dose spleen (30 g / kg Shenling Baizhu San), low-dose spleen (10 g / kg Shenling Baizhu San) Liver scattered Shenling Baizhu Powder), low-dose combination group (13.2 g / kg Chai Shu Shugan San Shen Ling Baizhu San), 9 in each group. NASH rat model was established by feeding with high-fat diet (10 mL / kg). After 16 w, rats in each group were anesthetized with 3% sodium pentobarbital, blood collected from the abdominal aorta, and blood lipid, Liver function, HE and oil red O staining liver tissue pathological changes; Real time Q-PCR detection of liver tissue TLR4 mRNA expression; Western blot detection of liver TLR4 protein expression levels. Results: Compared with the normal control group, the fatty degeneration of hepatic cells in the model group was obvious, the hepatocytes were enlarged, the infiltration of inflammatory cells in the lobes and portal area, the levels of TC, LDL-C, TG, TLR4 mRNA (P <0.01 or P <0.01). Compared with the model group, the expression of TLR4 mRNA and protein in the blood lipids, liver lipids and liver tissues of rats in each treatment group decreased to some extent (P <0.05 or P <0.01) High-dose effect is the most significant. Conclusion: Shuganjianpi Recipe may play an anti-NASH role by down-regulating the mRNA and protein expression of TLR4. TLR4 may be one of the effective anti-NASH targets of Shuganjianpi Decoction. Shuganjianpi Fang prevent NASH there may be a certain amount of dose-dependent relationship.