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)
凌琪华,女,博士,副主任医师,主要从事中医药防治肝病、急诊内科临床研究
乐敏,主任医师,硕士生导师;E-mail:lemin725@126.com
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凌琪华,吴荻,乐敏等.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[J].上海中医药大学学报,2022,36(S1):143-148.
LING Qihua,WU Di,LE min,et al.Study on mechanism of Cangju Qinggan Formula in treatment of non⁃alcoholic fatty liver based on “liver⁃bile acid⁃intestinal microecology”[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2022,36(S1):143-148.
凌琪华,吴荻,乐敏等.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[J].上海中医药大学学报,2022,36(S1):143-148. DOI: 10.16306/j.1008-861x.2022.S1.034.
LING Qihua,WU Di,LE min,et al.Study on mechanism of Cangju Qinggan Formula in treatment of non⁃alcoholic fatty liver based on “liver⁃bile acid⁃intestinal microecology”[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2022,36(S1):143-148. DOI: 10.16306/j.1008-861x.2022.S1.034.
目的,2,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,2,32 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,2,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,2,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 水平,并能改善NAFLD 大鼠病理状态以及脂质沉积状况。其主要通过改善群落物种数目,使高脂饮食喂养的NAFLD 大鼠紊乱的肠道菌群组成和结构逐渐恢复正常,从而改善NAFLD。
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,2,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,2,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 group presented diffuse vacuoles of different sizes and balloon-like changes, the infiltration of inflammatory cells in the portal area was obvious, and the hepatocytes were disordered with lots of lipid droplet of different sizes and dark color. All treatment groups could improve the damage degree of liver cells in NAFLD rats, with partially scattered vacuoles, and some liver cells returned to normal. The intestinal microflora of Cangju Formula groups were different from those of model group, and gradually approached to the state of the normal group.,Conclusion,2,Cangju Formula can reduce the levels of ALT, AST, TC, TG, LDL-C in serum of rats, and increase the level of HDL-C in serum of rats. It can also improve the pathological status and lipid deposition of NAFLD rats. And it improves NAFLD mainly by improving the number of community species and gradually restoring the composition and structure of disturbed intestinal flora in NAFLD rats fed with high-fat diet to normal.
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
Cangju Qinggan Formulanon-alcoholic fatty liver diseaseintestinal floramodel ratsresearch of traditional Chinese medicine
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