基于高通量测序分析凉血解毒方对HBV相关慢加急性肝衰竭患者肠道菌群的影响  

Effect of Liangxue Jiedu decoction on intestinal flora in patients with hepatitis B virus-related acute-on-chronic liver failure:An analysis based on high-throughput sequencing

作　　者：侯艺鑫[1] 张群 江宇泳[1] 于浩[1] 杨玉英[1] 王宪波[1] HOU Yixin;ZHANG Qun;JIANG Yuyong;YU Hao;YANG Yuying;WANG Xianbo(Center of Integrative Medicine,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China)

机构地区：[1]首都医科大学附属北京地坛医院中西医结合中心,北京100015

出　　处：《临床肝胆病杂志》2022年第6期1280-1287,共8页Journal of Clinical Hepatology

基　　金：北京市科学技术委员会“首都临床特色应用研究”资助项目(Z181100001718052);北京市医院管理中心“青苗”计划(QML20201802);北京市中医药科技发展资金项目(QN-2020-25);国家中医药管理局重大疑难疾病中西医临床协作试点项目(2018-6-4);国家中医药管理局区域中医诊疗中心(2019-3-18)。

摘　　要：目的观察凉血解毒方对HBV相关慢加急性肝衰竭(HBV-ACLF)患者肠道菌群的影响。方法选取2018年10月—2019年10月于北京地坛医院住院诊断为HBV-ACLF的患者,招募同期正常健康人群作为对照(HP组),通过高通量测序的方法筛选出HBV-ACLF患者与正常健康人群的菌群多样性和菌种的差异,从门、属水平筛选出两组的差异菌;利用体外模拟发酵实验,HBV-ACLF患者给予不同浓度(0、10%、50%、100%)凉血解毒方培养基培养24 h,从属分类水平比较分析HBV-ACLF用药组、HBV-ACLF未用药组及HP组菌群变化。正态分布的计量资料两组间比较采用t检验;非正态分布的计量资料两组间比较采用Mann-Whitney U检验。结果共纳入HBV-ACLF患者10例,其中HBV-ACLF用药组5例,未用药组5例;HP组15例。HBV-ACLF未用药组较HP组肠道菌群多样性及丰度下降。在门分类水平,HP组每个样本均以拟杆菌门和厚壁菌门为主;HBV-ACLF未用药组拟杆菌门明显减少,梭杆菌门、变形菌门、纤维杆菌门增多。在属分类水平上,HBV-ACLF未用药组瘤胃菌、布劳特菌属、真杆菌属较HP组明显减少,而副杆菌属、乳酸杆菌、梭杆菌、链球菌较HP组增多。体外模拟发酵实验结果显示,HBV-ACLF用药组瘤胃菌、毛螺菌属、拟杆菌、氏菌属相对丰度值较HBV-ACLF未用药组显著升高,而梭杆菌、变形菌较未用药组显著减少(P值均<0.05)。结论凉血解毒方能够调节肠道菌群紊乱,恢复肠道菌群多样性,增加优势菌,减少致病菌,这可能是其治疗HBV-ACLF重要的作用机制之一。Objective To investigate the effect of Liangxue Jiedu decoction on intestinal flora in patients with hepatitis B virus-related acute-on-chronic liver failure(HBV-ACLF).Methods The patients who were hospitalized and diagnosed with HBV-ACLF in Bei-jing Ditan Hospital from October 2018 to October 2019 were enrolled,and healthy individuals were enrolled as HP group.High-throughput sequencing was used to screen for the differences in bacterial diversity and species between HBV-ACLF patients and healthy individuals,and differentially expressed bacteria between the two groups were screened out at the phylum and genus levels.With the help of in vitro sim-ulated fermentation experiment,fecal samples were collected from the patients with HBV-ACLF and were then cultured in the medium con-taining different concentrations of Liangxue Jiedu decoction(0,10%,50%,and 100%)for 24 hours,and the changes in intestinal flora were analyzed and compared between the HBV-ACLF treatment group,the HBV-ACLF non-treatment group,and the HP group at the genus level.The t-test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups.Results A total of 10 HBV-ACLF patients were enrolled,with 5 in the HBV-ACLF treatment group and 5 in the HBV-ACLF non-treatment group,and there were 15 individuals in the HP group.Compared with the HP group,the HBV-ACLF non-treatment group had significant reductions in the diversity and abun-dance of intestinal flora.At the phylum level,Bacteroidetes and Firmicutes were mainly observed in the samples of the HP group,while the HBV-ACLF non-treatment group had a significant reduction in Bacteroidetes and significant increases in Fusobacteria,Proteobacteria,and Fibrobacteres.At the genus level,compared with the HP group,the HBV-ACLF non-treatment group had significant reductions in Rumi-nococcus,Blautia,and Eubacterium and significant increases in Parabacteroides,Lactobacillus,Fus

关 键 词：乙型肝炎病毒 慢加急性肝功能衰竭 中西医结合疗法 胃肠道微生物组 

分 类 号：R512.62[医药卫生—内科学] R575.3[医药卫生—临床医学]