基于HMGB1/TLR4/NF-κB通路探讨桃仁承气汤调节肠源性脓毒症大鼠肠道肌电活动及微环境稳态的作用机制  

Study on Mechanism of Taoren Chengqitang in Regulating Intestinal Myoelectrical Activity and Microenvironment Homeostasis in Intestinal Sepsis Rats Based on HMGB1/TLR4/NF-κB Pathway

作　　者：王丽辉 孙治霞[1] 索红亮[1] 郭艳青[1] 袁彬[1] 李宗尚 李莉[1] WANG Li-hui;SUN Zhi-xia;SUO Hong-liang;GUO Yan-qing;YUAN Bin;LI Zong-shang;LI Li(Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南中医药大学第二附属医院,郑州450002

出　　处：《中国实验方剂学杂志》2020年第12期78-84,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省中医药科学研究专项(2018ZY2050)。

摘　　要：目的:基于高迁移率族蛋白B1(HMGB1)/Toll样受体4(TLR4)/核转录因子-κB(NF-κB)通路研究桃仁承气汤调节肠源性脓毒症大鼠肠道肌电活动及微环境稳态的作用机制。方法:60只SD大鼠随机分为假手术组、模型组、甘草酸(HMGB1抑制剂,0.03 g·kg^-1)组、桃仁承气汤(10 g·kg^-1)组、甘草酸+桃仁承气汤(0.03 g·kg^-1+10 g·kg^-1)组,每组12只。除假手术组外,其余各组建立肠源性脓毒症大鼠模型,各组分别以药物处理后,采用苏木素-伊红(HE)染色检测各组大鼠小肠黏膜组织病理改变,比较黏膜厚度、绒毛高度变化;采用试剂盒检测各组大鼠小肠黏膜组织分泌型免疫球蛋白A(sIgA)含量,血清二胺氧化酶(DAO)及D-乳酸水平;检测各组大鼠肠道肌电活动,比较小肠平滑肌慢波频率、慢波振幅;检测各组大鼠肠道菌群,比较大肠埃希菌、双歧杆菌和乳酸杆菌含量;采用蛋白免疫印迹法(Western blot)检测小肠组织中HMGB1/TLR4/NF-κB通路蛋白HMGB1,TLR4,MyD88,NF-κB p65表达情况。结果:与假手术组比较,模型组大鼠小肠黏膜组织绒毛高度、黏膜厚度,sIgA含量,平滑肌慢波频率及振幅、肠道双歧杆菌及乳酸杆菌含量显著降低,血清DAO及D-乳酸水平、肠道大肠埃希菌含量、小肠组织HMGB1,TLR4,MyD88及NF-κB p65蛋白表达明显升高(P<0.05);与模型组比较,桃仁承气汤组、甘草酸组、甘草酸+桃仁承气汤组大鼠小肠黏膜组织绒毛高度、黏膜厚度,sIgA含量、平滑肌慢波频率及振幅、肠道双歧杆菌及乳酸杆菌含量升高,血清DAO及D-乳酸水平、肠道大肠埃希菌含量、小肠组织HMGB1,TLR4,MyD88及NF-κB p65蛋白表达降低(P<0.05);与桃仁承气汤组、甘草酸组分别比较,甘草酸+桃仁承气汤组大鼠小肠黏膜组织绒毛高度、黏膜厚度,sIgA含量、平滑肌慢波频率及振幅、肠道双歧杆菌及乳酸杆菌含量升高,血清DAO及D-乳酸水平、肠道大肠埃希菌含量、小肠组织HMGB1,TLR4,Objective:To study the mechanism of Taoren Chengqitang in regulating intestinal myoelectric activity and microenvironment homeostasis in intestinal sepsis rats based on high mobility group protein 1(HMGB1)/Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)pathway.Method:The 60 SD rats were randomly divided into sham operation group,model group,glycyrrhizic acid(HMGB1 inhibitor,0.03 g·kg^-1)group,Taoren Chengqitang group(10 g·kg^-1),glycyrrhizic acid+Taoren Chengqitang group(0.03 g·kg^-1+10 g·kg^-1),with 12 rats in each group.Except the sham operation group,the other groups established intestinal sepsis rat models,each group was treated with medicine,hematoxylin-eosin(HE)staining was used to detect the histopathological changes of small intestinal mucosa in rats of each group,the changes of mucosal thickness and villus height were compared,the levels of secretory immunoglobulin A(sIgA),diamine oxidase(DAO)and D-lactic acid in intestinal mucosa of rats were detected by kit,the intestinal myoelectrical activity of rats in each group was measured,the slow wave frequency and amplitude of small intestinal smooth muscle were compared,the intestinal flora of rats in each group was detected,the contents of E.coli,Bifidobacterium and Lactobacillus were compared,and the expressions of HMGB1/TLR4/NF-κB pathway proteins HMGB1,TLR4,MyD88 and NF-κB p65 in small intestinal tissues were detected by Western blot.Result:Compared with sham operated group,the villus height,mucosal thickness,sIgA content,slow wave frequency and amplitude of smooth muscle,Bifidobacterium and Lactobacillus contents in intestinal mucosa of model group rats were significantly decreased,and serum DAO and D-lactic acid levels,intestinal E.coli content,intestinal tissue HMGB1,TLR4,MyD88 and NF-κB p65 proteins were significantly increased(P<0.05).Compared with the model group,the villus height,mucosal thickness,sIgA content,slow wave frequency and amplitude of smooth muscle,Bifidobacterium and Lactobacillus contents in intestinal mucosa of the Taore

关 键 词：桃仁承气汤 高迁移率族蛋白B1(HMGB1)/Toll样受体4(TLR4)/核转录因子κB(NF-κB)通路 肠源性脓毒症 肠道肌电活动 微环境稳态 

分 类 号：R2-0[医药卫生—中医学] R289