摘要
目的探讨扶脾柔肝方及拆方对肝纤维化(hepatic fibrosis,HF)大鼠肝组织线粒体自噬相关蛋白表达的影响。方法48只SD大鼠随机分为空白组、模型组、秋水仙碱组(0.2 mg·kg^(-1))、扶脾柔肝方组(8.54 g·kg^(-1))、扶脾方组(4.58 g·kg^(-1))、柔肝方组(3.96 g·kg^(-1))。采用四氯化碳腹腔注射和乙醇灌胃复制HF大鼠模型,灌胃给药4周。检测大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST);HE、Masson染色观察病理变化情况;电镜观察大鼠肝组织超微结构;Western Blot和qRT-PCR方法检测大鼠肝组织PINK1、Parkin、Beclin 1和LC3-Ⅱ基因及蛋白表达水平。结果与正常组比较,模型组的HF程度明显,胶原容积分数明显增加(P<0.01),电镜发现自噬小体增多,线粒体结构损害,大鼠血清ALT、AST明显升高(P<0.01),肝组织PINK1、Parkin、Beclin 1和LC3-ⅡmRNA及蛋白表达上调(P<0.01);与模型组比较,扶脾柔肝方及拆方HF程度有减轻,胶原容积分数明显下降(P<0.01),自噬小体减少,线粒体损害减轻,全方及扶脾方组大鼠血清ALT、AST明显降低(P<0.01),肝组织PINK1、Parkin、Beclin 1和LC3-ⅡmRNA及蛋白表达下调(P<0.01,P<0.05)。结论扶脾柔肝方及拆方抗肝纤维化,全方可发挥最佳疗效,其机制可能与下调PINK1、Parkin、Beclin 1和LC3-Ⅱ基因及蛋白的表达,干预细胞线粒体自噬水平有关。
Objective To explore the effects of Fupi Rougan Formula(扶脾柔肝方)and its splited prescriptions on the expressions of mitochondrial autophagy-related proteins in liver tissues of hepatic fibrosis(HF)rats.Methods Forty-eight SD rats were randomly divided into blank group,model group,colchicine group(0.2 mg·kg^(-1)),Fupi Rougan Formula group(8.54 g·kg^(-1))group,and Fupi Formula(扶脾方)group(4.58 g·kg^(-1)),Rougan Formula(柔肝方)group(3.96 g·kg^(-1)).The HF rat model was replicated by intraperitoneal injection of carbon tetrachloride and ethanol gavage for 4 weeks.The levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the serum were observed.HE and Masson staining methods were used to observe pathological changes.The ultrastructure of rats liver tissue was observed by electron microscope.Western blot and qRT-PCR methods were used to detect PINK1,Parkin,Beclin 1 and LC3-Ⅱgene and protein expression levels in rats liver tissues.Results Compared with that of the normal group,the degree of HF in the model group was obvious.The volume fraction of collagen was increased significantly(P<0.01).The electron microscope found that autophagosomes increased,mitochondrial structure was damaged,and the levels of ALT and AST in serum increased significantly(P<0.01).The expressions of PINK1,Parkin,Beclin1 and LC3-ⅡmRNA and protein in liver tissues were up-regulated(P<0.01).Compared with the model group,Fupi Rougan Formula and the splited prescriptions reduced HF degree and lowered the collagen volume fraction significantly(P<0.01).The autophagosomes were reduced,and mitochondrial damage was alleviated.ALT and AST were significantly reduced(P<0.01),and expressions of PINK1,Parkin,Beclin1,and LC3-II mRNA and protein in liver tissue were down-regulated(P<0.01,P<0.05).Conclusion Fupi Rougan Formula and its splited prescriptions can exert the best effect on hepatic fibrosis.The mechanism may be related to down-regulating the expressions of PINK1,Parkin,Beclin1 and LC3-Ⅱgenes and proteins,and interfering with the level of mitochondrial autophagy.
作者
安祯祥
何远利
王芳
唐东昕
王敏
粟周云
AN Zhenxiang;HE Yuanli;WANG Fang;TANG Dongxin;WANG Min;SU Zhouyun(The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,Guizhou,China)
出处
《中华中医药学刊》
CAS
北大核心
2022年第2期11-15,I0025-I0028,共9页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金(81760865,81460726)
贵州省卫生与计划生育委员会科学技术基金(gzwjkj2015-1-058)。
