枯草芽孢杆菌通过TGR5/TRPA1信号通路调节慢性传输型便秘小鼠的肠蠕动  

Bacillus Subtilis Regulates Intestinal Peristalsis in Mice with Slow Transit Constipation Through the TGR5/TRPA1 Signaling Pathway

作　　者：陈振海 冯江毅 胡淞 华烨[1] 马莎英 付伟杰 杨钱[1] 张鑫 CHEN Zhenhai;FENG Jiangyi;HU Song;HUA Ye;MA Shaying;FU Weijie;YANG Qian;ZHANG Xin(Department of General Surgery,Chongqing Emergency Medical Center/Chongqing University Central Hospital,Chongqing 400014,China)

机构地区：[1]重庆市急救医疗中心普通外科/重庆大学附属中心医院,重庆400014

出　　处：《中国现代应用药学》2022年第7期878-884,共7页Chinese Journal of Modern Applied Pharmacy

基　　金：重庆市渝中区基础研究与前沿探索项目(20190140)。

摘　　要：目的探究枯草芽孢杆菌对慢性传输型便秘(slow transit constipation,STC)小鼠5-HT释放及TGR5/TRPA1信号通路的影响,以明确其对STC的治疗作用及机制。方法24只♂C57BL/6小鼠分为对照组、模型组、枯草芽孢杆菌治疗组和INT-777组。复方地芬诺酯用于构建STC小鼠模型,枯草芽孢杆菌治疗组和INT-777组于复方地芬诺酯灌胃1 h后分别灌胃枯草芽孢杆菌悬液和INT-777,连续14 d。实验期间统计小鼠24 h排便量、粪便含水量和肠道推进率;HE染色观察结肠病变程度;ELISA和免疫荧光检测结肠5-HT的表达;Western blotting检测结肠TGR5/TRPA1通路蛋白的表达。结果与对照组相比,模型组小鼠24 h排便量、粪便含水量和肠道推进率均显著降低(P<0.01或P<0.001),表现出肠蠕动障碍和结肠病理损伤;枯草芽孢杆菌和INT-777显著改善了肠蠕动障碍和结肠病理损伤。另外,模型组小鼠5-HT的释放和TGR5/TRPA1通路被抑制,而枯草芽孢杆菌和INT-777显著增加了STC小鼠结肠组织中5-HT的表达(P<0.05或P<0.001),激活了TGR5/TRPA1通路。结论枯草芽孢杆菌可能通过上调TGR5/TRPA1通路并促进肠嗜铬细胞释放5-HT调节STC小鼠肠蠕动,对STC发挥较好的治疗作用。OBJECTIVE To explore the effects of Bacillus subtilis on the release of 5-HT and the TGR5/TRPA1 signaling pathway in mice with slow transit constipation(STC),so as to clarify the therapeutic effect and mechanism of Bacillus subtilis on STC.METHODS Twenty-four male C57BL/6 mice were divided into control group,model group,Bacillus subtilis group and INT-777 group.STC mouse model was established by compound diphenoxylate,and Bacillus subtilis suspension and INT-777 were gavaged after 1 h compound diphenoxylate treatment,respectively,for 14 d.The 24 h defecation volume,fecal water content and intestinal transport rate of mice were counted during the experiment.HE staining was used to observe the histological morphology and pathological degree of colon.The expression of 5-HT in colon was detected by ELISA and immunofluorescence,and the protein expressions of TGR5/TRPA1 pathway were detected by Western blotting.RESULTS It was found that compared with the control group,the 24 h defecation volume,fecal water content and intestinal transit rate in the model group were significantly decreased(P<0.01 or P<0.001),showing intestinal peristalsis disorder and pathological lesions of the colon.Bacillus subtilis and INT-777 significantly improved intestinal peristalsis and pathological damage of colon.In addition,the release of 5-HT(P<0.05 or P<0.001)and the TGR5/TRPA1 signaling pathway in the model group were inhibited,while Bacillus subtilis and INT-777 significantly increased the expression of 5-HT and activated the TGR5/TRPA1 pathway in STC mice.CONCLUSION Bacillus subtilis regulates intestinal peristalsis of STC mice by upregulating the TGR5/TRPA1 signaling pathway and promoting the release of 5-HT from enterochromaffin cells,thus playing a good role in the treatment of STC.

关 键 词：枯草芽孢杆菌 肠蠕动 肠嗜铬细胞 5-HT TGR5/TRPA1信号通路 慢性传输型便秘 

分 类 号：R965.1[医药卫生—药理学]