基于“肝脏-胆汁酸-肠道微生态”的苍菊清肝方治疗非酒精性脂肪肝的机制研究  

Study on mechanism of Cangju Qinggan Formula in treatment of non-alcoholic fatty liver based on “liverbile acid-intestinal microecology”

作　　者：凌琪华[1] 吴荻 乐敏[2] 陈建杰[2] LING Qihua;WU Di;LE min;CHEN Jianjie(Department of Emergency,Shuguang Hospital Affiliate to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Hepatology,Shuguang Hospital Affiliate to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院急诊科,上海201203 [2]上海中医药大学附属曙光医院肝科,上海201203

出　　处：《上海中医药大学学报》2022年第S01期143-148,共6页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：上海市“科技创新行动计划”医学创新研究专项项目(21Y11920500);上海中医药大学预算内项目(2020LK044)

摘　　要：目的:探索高脂饮食诱导的非酒精性脂肪性肝病(NAFLD)模型小鼠的“肝脏-胆汁酸-肠道微生态轴”失衡的病理机制,验证苍菊清肝方对“肝脏-胆汁酸-肠道微生态轴”失衡的调节,进一步阐明苍菊方治疗NAFLD的机制。方法:32只雄性SPF级SD大鼠随机分为空白组(N,n=5)和高脂饮食组(HFD,n=27)。空白组5只给予普通饲料,高脂饲料组以高脂饲料(胆固醇1%,猪油10%,蔗糖20%)喂养,诱导NAFLD模型。饲养8周后,高脂饮食组造模成功的25只大鼠再随机分为模型组(HFDM,n=5)、益生菌组(YSJ,n=5)、苍菊方高剂量组(CJFH,n=5)、苍菊方中剂量组(CJFM,n=5)、苍菊方低剂量组(CJFL,n=5)。利用全自动生化仪检测各组大鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、三酰甘油(TG)、总胆固醇(TC)含量。HE染色观察大鼠肝组织病变程度以及脂质沉积情况。用Illumina高通量测序法,对大鼠粪便样本总DNA的16Sr RNA的V3~V4区进行基因测序以及生物信息学分析。结果:与空白组比较,模型组血清ALT、AST升高,TC、TG、LDL-C升高,HLD-C降低,差异均有统计学意义(P<0.05)。与模型组比较,益生菌组、苍菊方各剂量组大鼠血清ALT、AST、TC下降(P<0.05),苍菊方中高剂量组大鼠血清大鼠血清TG下降(P<0.05),益生菌组、苍菊方中高剂量组大鼠血清HLD-C升高,苍菊方高剂量组大鼠血清LDL-C下降(P<0.05)。HE染色显示,模型组大鼠肝脏中肝细胞出现弥漫性、大小不等的空泡,出现气球样变,汇管区炎症细胞浸润明显,肝细胞排列紊乱可见大量大小脂滴,颜色深。各给药组均能改善NAFLD大鼠肝细胞内损伤程度,存在部分散在的空泡,部分肝细胞恢复正常。苍菊方各剂量组肠道菌群与模型组有不同程度的差异,逐渐接近正常组状态。结论:苍菊方能降低大鼠血清中ALT、AST、TC、TG、LDL-C水平,升高大鼠血清中HDL-C�Objective:To explore the pathological mechanism of the imbalance of“liver-bile acid-intestinal microbiological axis”in non-alcoholic fatty liver disease(NAFLD)model mice induced by high-fat diet,verify the regulation of Cangju Qinggan Formula on the imbalance of“liver-bile acid-intestinal microbiological axis”,and further illustrate the mechanism of Cangju Formula in the treatment of NAFLD.Methods:Thirty-two SPF male SD rats were randomly divided into blank group(N,n=5)and high-fat diet group(HFD,n=27).Five mice in blank group were only given ordinary diet,and high-fat diet group was fed with hrgh-fat diet(cholestenol 1%,lard 10%,sucrose 20%)to induce NAFLD model.After 8 weeks of feeding,25 rats in the high-fat diet group were randomly divided into model group(HFDM,n=5),probiotic group(YSJ,n=5),Cangju Formula high-dose group(CJFH,n=5),Cangju Formula medium-dose group(CJFM,n=5),Cangju Formula low-dose group(CJFL,n=5).The contents of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),triglyceride(TG)and total cholesterol(TC)in serum of rats in each group were measured by automatic biochemical instrument.The degree of liver lesion and lipid deposition in rats were observed by HE staining.Using Illumina high-throughput sequencing method,the V3~V4 region of 16sr RNA of total DNA of rat fecal samples was sequenced and bioinformatics was analyzed.Results:Compared with blank group,serum ALT and AST increased,TC,TG and LDL-C increased,and HLD-C decreased in model group,the differences were statistically significant(P<0.05).Compared with the model group,serum ALT,AST and TC decreased in the probiotic group and Cangju Formula groups(P<0.05),serum TG decreased in Cangju Formula medium and high dose group(P<0.05),serum HDD-C increased in the probiotic group,Cangju Formula medium and high dose group,and serum LDL-C decreased in Cangju Formula high-dose group(P<0.05).HE staining showed that hepatocytes in the model gr

关 键 词：苍菊清肝方 非酒精性脂肪性肝病 肠道菌群 模型大鼠 中药研究 

分 类 号：R285.5[医药卫生—中药学]