疏肝健脾方药对非酒精性脂肪性肝病大鼠肝组织LXRαmRNA及蛋白表达的影响

目的 探讨疏肝健脾方药对非酒精性脂肪性肝病( NAFLD)大鼠肝组织LXRα mRNA及蛋白表达的影响.方法 选用SD大鼠55只,随机分为正常组、模型组、疏肝组(灌服3.2g·kg-1·d-1剂量的柴胡疏肝散)、健脾组(灌服10.0g·kg -1·d-1剂量的参苓白术散)、综合组(灌服11.9g·kg-1·d-1剂量的柴胡疏肝散和参苓白术散合方),模型组15只,其余各组10只.采用灌饲高脂肪乳剂(10 mL/kg)的方法复制大鼠NAFLD实验动物模型,给药8 w后处死动物,腹主动脉采血,用全自动生化分析仪检测血脂及肝功;常规HE染色观察肝组织病理变化;RT-PCR方法检测肝组织LXRα mRNA的表达;免疫组织化学方法检测肝组织LXRα蛋白的表达.结果 与正常组相比,模型组大鼠肝细胞脂肪变性明显,血脂及肝功均有不同程度的升高(P＜0.05,P＜0.01),大鼠肝组织LXRα mRNA及蛋白表达明显升高(P＜0.01);各给药组血脂及肝功和肝组织LXRαmRNA及蛋白的表达均较模型组显著降低(P＜0.05,P＜0.01),其中以健脾组下降最为明显.结论 疏肝健脾方药对高脂饮食诱导的大鼠NAFLD有较好的治疗作用,其机制可能与其下调肝脏LXRα的表达有关.     

Effects of different therapeutic methods and typical recipes of Chinese medicine on activation of c-Jun N-terminal Kinase in Kupffer cells of rats with fatty liver disease

<正>Objective:To observe the effects of different therapeutic methods and the recipes of Chinese medicine(CM) on the activation of c-Jun N-terminal kinase(JNK) in Kupffer cells of rats with fatty liver disease and to explore the mechanisms of these therapeutic methods.Methods:By using a random number table,98 rats were randomly divided into 7 groups:control group,model group,and 5 treatment groups,including soothing Liver(Gan) recipe group,invigorating Spleen(Pi) recipe group,dispelling dampness recipe group,promoting blood recipe group,and complex recipe group.Rats in the control group were fed with normal food and distilled water by gastric perfusion,while rats in the model group were fed with high-fat food and distilled spirits by gastric perfusion.Rats in the 5 treatment groups were fed with high-fat food and corresponding recipes by gastric perfusion.Twelve weeks later,all rats were sacrificed and liver tissues were stained for pathohistological observation.Kupffer cells were isolated from livers of rats to evaluate JNK and phospho-JNK expressions by Western blotting.Results:The grade of hepatic steatosis was higher in the model group than the control group(P0.05).Compared with the model group,the grade of fatty degeneration in soothing Liver recipe group and invigorating Spleen recipe group were significantly ameliorated(P0.05).Expressions of JNK and phospho-JNK in Kupffer cells were significantly higher in the model group than those in the control group(P0.05,P0.01).Compared with the model group,expressions of JNK in all treatment groups decreased,especially in invigorating Spleen recipe group and promoting blood recipe group(P0.05).Compared with the model group,expressions of phospho-JNK in all treatment groups declined significantly(P0.01),especially in soothing Live recipe group and invigorating Spleen recipe group. Conclusions:The high expressions of JNK and phospho-JNK in Kupffer cells might play an important role in the pathogenesis of fatty liver disease in rats.The recipes of CM,especially invigorating Spleen recipe and soothing Liver recipe,might protect liver against injury by reducing the total JNK protein content and inhibiting the activation of JNK protein in Kupffer cells of fatty liver model rats,which showed beneficial effects on fatty liver disease.     

浅论经前期综合征从肝脾治疗

经前期综合征（Premenstrual Syndrome,PMS）是育龄期妇女发病率较高的疾病之一。笔者通过对中医经典著作的整理、研究认为对该病的治疗应当从肝脾二脏入手,采用疏肝健脾之法取得较好疗效。     

Effect of berberine on expressions of uncoupling protein-2 mRNA and protein in hepatic tissue of non-alcoholic fatty liver disease in rats

Objective  

To observe the effect of berberine on uncoupling protein-2 (UCP2) mRNA and protein expressions in the hepatic tissue of non-alcoholic fatty liver disease (NAFLD) in rats, and to explore the molecular mechanism.     

三七总皂甙对脂肪变性L02肝细胞TG水平及SREBP-1c mRNA表达的影响

目的观察三七总皂甙（PNS）对脂肪变性肝细胞的降脂作用及机制。方法采用50％小牛血清诱导L02肝细胞48h建立L02肝细胞脂肪变性模型,分为模型组、自然恢复组、PNS低剂量组、PNS高剂量组,并设正常组。除模型组继续予含50％小牛血清的1640培养基培养外,余组均改予含10％小牛血清培养。PNS低、高剂量组分别予10、50μg／ml PNS作用。药物作用24h后,油红O染色观察肝细胞内脂滴变化,全自动生化仪检测TG水平,RT—PCR法检测固醇元件结合蛋白-1C（SREBP—lc mRNA）表达。结果与自然恢复组比较,PNS各治疗组细胞内脂滴少于其他组,低剂量组更为明显;TG水平明显低于其他各组（P〈0．05）,低剂量组下降更为显著（P〈0．01）;SREBP—lc mRNA的表达量均有下降,低剂量组更为明显（P〈0．05）。结论PNS能显著降低脂肪变性肝细胞内TG水平,减轻肝细胞脂肪变性;机制可能与下调SREBP-1cm RNA的表达有关。     

从痰湿体质探讨非酒精性脂肪性肝病的遗传易感性及防治

非酒精性脂肪性肝病是遗传-环境-代谢应激性疾病,遗传易感性与其发生密切相关。痰湿体质的先天禀赋因素具有遗传易感性基础,从痰湿体质角度理解非酒精性脂肪性肝病发病易感性,加深了对该病中医病因、病机的认识,以辨体质为基础的辨证论治对治疗具有指导意义。     

疏肝健脾方药对非酒精性脂肪性肝病大鼠肝组织UCP2 mRNA及蛋白表达的影响

目的观察疏肝健脾方药对非酒精性脂肪性肝病（non—alcoholic fatty liver disease,NAFLD）大鼠肝组织解偶联蛋白2（uncoupling protein 2,UCP2）mRNA及蛋白表达的影响,初步探讨疏肝健脾方药治疗NAFLD的作用机制。方法采用高脂饲料喂养12周复制雄性SD大鼠NAFLD模型,将模型大鼠分为疏肝纽、健脾组、疏肝健脾综合组（综合组）、三七脂肝丸组（三七组）、模型组,各组给予相应治疗8周。测定各组大鼠血清总胆固醇、三酰甘油、低密度脂蛋白胆固醇和高密度脂蛋白胆固醇含量。采用RT—PCR方法检测肝组织UCP2 mRNA的表达,免疫组织化学方法检测肝组织中UCP2蛋白活性变化。结果与正常组比较,模型组大鼠肝组织中UCP2 mRNA表达水平显著升高（P〈0．05）;与模型组比较,疏肝组、健脾组和综合组UCP2 mRNA表达水平显著下降（P〈0．05）。模型组肝组织UCP2蛋白表达水平显著升高;与模型组比较,各用药组UCP2蛋白表达水平均显著下调（P〈0．01）。结论疏肝健脾方药可使NAFLD大鼠肝组织中UCP2基因和蛋白表达水平降低。     

疏肝健脾方药对非酒精性脂肪性肝病大鼠肝组织脂肪酸合成酶mRNA及蛋白表达的影响

目的:观察疏肝健脾方药对非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)大鼠肝组织脂肪酸合成酶(fatty acid synthase,FAS)mRNA及蛋白表达的影响,探讨疏肝健脾方药抗大鼠NAFLD的作用机制。方法:雄性SD大鼠,分为正常组、模型组、健脾组、疏肝组、综合组。使用高脂肪乳剂灌胃造模,治疗组给予相应药物灌胃,8周后处死大鼠,检测血清转氨酶、血脂和肝脂水平;HE染色观察肝组织病理变化;RT-PCR和免疫组织化学方法检测肝组织FAS基因和蛋白的表达。结果:与模型组相比,各药物干预组大鼠血清AST、TC、TG含量和肝组织TC含量均显著下降(P<0.05,P<0.01),健脾组和疏肝组肝组织TG含量显著下降(P<0.05);光镜观察各药物均能改善大鼠肝细胞脂肪变性,以疏肝组更为显著;各用药组FAS mRNA表达水平和蛋白的表达均有下降(P<0.05),其中疏肝组表达的水平最低。结论:疏肝健脾方药能显著改善NAFLD大鼠脂质代谢紊乱,减轻肝损伤,具有良好的抗NAFLD的作用。其机制可能与下调FAS mRNA及蛋白表达有关。