基于胆汁酸代谢组学探讨复方大黄煎剂保留灌肠对轻微型肝性脑病大鼠模型的治疗作用  

Therapeutic effect of retention enema with compound rhubarb decoction on a rat model of minimal hepatic encephalopathy based on bile acid metabolomics

作　　者：杜沅沁 王萌[2] 黄国初[2] 姚春[1] 钟瑞熙 黄良江 徐健 黄晶晶[2] 谭钦文 毛德文[2] DU Yuanqin;WANG Meng;HUANG Guochu;YAO Chun;ZHONG Ruixi;HUANG Liangjiang;XU Jian;HUANG Jingjing;TAN Qinwen;MAO Dewen(Graduate School,Guangxi University of Chinese Medicine,Nanning 530000,China;Department of Spleen and Stomach Disease,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)

机构地区：[1]广西中医药大学研究生院,南宁530000 [2]广西中医药大学第一附属医院脾胃科,南宁530023

出　　处：《临床肝胆病杂志》2023年第10期2348-2357,共10页Journal of Clinical Hepatology

基　　金：国家自然科学基金(81804019);广西自然科学基金(2020GXNSFBA297033,2022GXNSFAA035460,2022GXNSFBA035485,2018GXNSFBA050041);广西研究生教育创新计划资助项目(YCBXJ2021013,YCXJ2021056);中医壮瑶治疗优势病种研究项目(GZBZ22-3)。

摘　　要：目的 观察大黄煎剂保留灌肠对轻微型肝性脑病(MHE)大鼠模型的治疗作用,并基于胆汁酸(BA)代谢组学探讨其作用机制。方法 将55只雄性SD大鼠随机分为空白组(NC组,n=10)、肝性脑病组(HE组,n=15)、轻微型肝性脑病组(MHE组,n=15)和MHE大黄煎剂治疗组(MHEY组,n=15)。腹腔注射CCl4和硫代乙酰胺(TAA)诱导MHE、HE大鼠模型,给药2周后处死。检测血清AST、ALT、ALP、TBil、总胆汁酸(TBA)和血氨的含量;取结肠内容物检测pH值;取肝组织和脑组织进行HE染色观察大鼠肝组织病理形态学改变;取胆汁用LC-MS进行BA靶向代谢组学分析。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 与NC组比较,HE组、MHE组寻台潜伏期(造模后、用药后)显著增加,穿台次数显著减少(P值均<0.05);与MHE组相比,MHEY组寻台潜伏期(用药后)显著降低而穿台次数显著增加,HE组寻台潜伏期显著增加而穿台次数显著减少(P值均<0.05)。与NC组比较,HE组和MHE组AST、ALT、ALP、TBil、TBA、血氨及结肠pH值显著增加(P值均<0.05);与MHE组相比,MHEY组AST、ALT、ALP、TBil、TBA及血氨及结肠pH值减少(P值均<0.05),HE组AST、ALT、ALP、TBil、TBA、血氨及结肠pH值增加(P值均<0.05)。MHE组TBA、初级BA和次级BA均低于NC组(P值均<0.05);HE组TBA和初级BA低于MHE组(P值均<0.05);MHEY组TBA、初级BA高于MHE组(P值均<0.05)。MHE组与NC组对比,GCDCA、GUDCA、GHDCA、TCDCA、TUDCA、GLCA和TLCA减少(P值均<0.05),γ-MCA、THCA、7-KDCA、AlloLCA、α-MCA增加(P值均<0.05)。MHEY组与MHE组对比,THDCA、TMCA、TCDCA、TUDCA和TLCA增加(P值均<0.05)。结论 大黄煎剂保留灌肠可通过调节BA肠-肝循环改善CCl4和TAA诱导的MHE大鼠模型肝损伤和认知功能,其作用机制可能与牛磺酸结合BA合成增加有关。Objective To investigate the therapeutic effect of rhubarb decoction(RD)retention enema on a rat model of minimal hepatic encephalopathy (MHE) and its mechanism of action based on bile acid (BA) metabolomics. Methods A total of 55 male Sprague-Dawley rats were randomly divided into blank group (NC group with 10 rats), hepatic encephalopathy group (HE group with 15 rats), MHE group with 15 rats, and MHE+rhubarb decoction treatment group (MHEY group with 15 rats). Intraperitoneal injection of carbon tetrachloride (CCl4) and thioacetamide (TAA) was performed to establish a rat model of MHE or HE, and the rats were sacrificed after 2 weeks of administration. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin (TBil), and total bile acid (TBA) and the concentration of blood ammonia were measured;the colonic contents were collected to measure pH value;liver and brain tissue samples were collected, and HE staining was used to observe the histopathological changes of the liver;the bile was collected, and liquid chromatography-mass spectrometry was used to perform BA-targeted metabolomics analysis. Continuous data were expressed as mean±standard deviation;a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results Compared with the NC group, the HE group and the MHE group had a significant increase in searching platform latency (after modelling and after administration) and a significant reduction in the number of platform crossings (all P<0.05);compared with the MHE group, the MHEY group had a significant reduction in searching platform latency (after administration) and a significant increase in the number of platform crossings, and the HE group had a significant increase in searching platform latency and a significant reduction in the number of platform crossings (all P<0.05). Compared with the NC group, the HE group an

关 键 词：肝性脑病 大黄煎剂 胆汁酸类 

分 类 号：R285.5[医药卫生—中药学]