逍遥散加味颗粒对大鼠肝纤维化TGF-β1/Smad通路的影响

Effect of Xiaoyao SAN Jiawei Granule on liver fibrosis through the TGF-β1/Smad pathway

目的:探讨逍遥散加味颗粒通过转化生长因子-β1(TGF-β1)/Smad通路改善肝纤维化的作用与机制。方法:将78只雌雄各半的SD大鼠随机分为正常组(n=12)和模型组(n=66)。模型组采用四氯化碳皮下注射,喂养高脂饲料造模9周后,将其随机分为对照组(n=12)、高剂量组(n=13)、低剂量组(n=13)、复方鳖甲组(n=13)及扶正化瘀组(n=12)。正常组与模型组不进行干预;高剂量组用逍遥散加味颗粒灌胃16 g•kg^(-1)•d^(-1);低剂量组用逍遥散加味颗粒灌胃8 g•kg^(-1)•d^(-1);复方鳖甲组采用复方鳖甲软肝片灌胃0.5 g•kg^(-1)•d^(-1);扶正化瘀组采用扶正化瘀胶囊灌胃0.5 g•kg^(-1)•d^(-1),1次/d,连续4周。末次给药禁食8 h后,大鼠进行目内眦取血,采用酶联免疫法(ELISA)测定血浆TGF-β1的含量;禁食24 h后,麻醉,固定,解剖肝组织,苏木素-伊红(HE)染色观察肝脏组织的变化情况,蛋白质免疫印迹检测肝组织中TGF-β1、Smad2蛋白的表达情况,并用RT-qPCR检测肝组织中TGF-β1 mRNA的表达情况。结果:与正常组对比,对照组肝脏组织可见肝纤维化改变,肝脏组织中的TGF-β1、Smad2表达水平上升;其余各治疗组肝脏组织肝纤维化改变较轻,高、低剂量治疗组肝脏组织病理改变明显,较扶正化瘀组及复方鳖甲组轻,且组织中TGF-β1、Smad2表达水平低于对照组、扶正化瘀组及复方鳖甲组。结论:逍遥散加味颗粒可以通过抑制TGF-β1/Smad2通路,改善大鼠肝纤维化病变及逆转肝纤维化。

Objective:To investigate the effect and mechanism of Xiaoyao SAN Jiawei Granule on liver fibrosis through the transforming growth factor-β1(TGF-β1)/Smad pathway.Methods:A total of 78 SD rats were randomly assigned to the normal group(n=12)and the model group(n=66).The model group was administered subcutaneous injections of carbon tetrachloride and fed a high-fat diet for a duration of 9 weeks.Subsequently,they were randomly divided into the following groups:control group(n=12),high-dose group(n=13),low-dose group(n=13),compound Biejia group(n=12),and Fuzheng Huayu group(n=12).No intervention was performed in the normal group and model group,while the other groups received corresponding drug doses via intragastric administration.The high-dose group was administered 16 g/(kg•d)of Xiaoyao SAN Jiawei granules,the low-dose group was given 8 g/(kg•d)of Xiaoyao SAN Jiawei granules,the compound Biejia group was administered with compound Biejia Ruangan tablet at a dose of 0.5 g/(kg•d),and the Fuzheng Huayu group was administered with Fuzheng Huayu capsule at a dose of 0.5 g/(kg•d)by gavage,once a day for 4 weeks.After fasting for 8 h after the last administration of the drug,the rats were collected from the inner canthus,and the plasma TGF-β1 content was determined by enzyme-linked immunoassay(ELISA).After fasting for 24 h,the liver tissues were anesthetized,immobilized,dissected,and stained with hematoxylin-eosin(HE)to observe the changes in the liver tissues.The expression of TGF-β1 and Smad2 proteins in the liver tissues was detected by western blot,and the expression of TGF-β1 mRNA in the liver tissues was detected by real-time quantitative fluorescent PCR.Results:Compared with the normal group,liver fibrosis was observed in the control group,and the expression levels of TGF-β1 and Smad2 in liver tissues were increased.The liver fibrosis changes in the other treatment groups were mild,and the liver histological changes in the high and low dose treatment groups were lighter than that in the Fuzheng Huayu group and the compound Biejia group.The expression levels of TGF-β1 and Smad2 in the liver tissues were lower than those in the control group,the Fuzheng Huayu group and the compound Biejia group.Conclusion:Xiaoyao SAN Jiawei Granule can improve liver fibrosis in rats and reversing liver fibrosis through inhibit TGF-β1/Smad2 pathway.