益肠通秘汤对便秘小鼠菌群多...Occludin表达的影响_黄玲.pdf

益肠通秘汤对便秘小鼠菌群多样性及肠道Occludin 表达的影响黄玲1，潘姣2，石镇阳1，林爱珍3(1 湖北中医药大学中医临床学院，湖北 武汉 430065;2 岳阳市人民医院，湖南 岳阳414100;3 湖北省中医院，湖北 武汉 430065)摘要 目的探讨益肠通秘汤对便秘小鼠菌群多样性及肠道紧密连接蛋白(Occludin)表达的影响。方法将 40 只 Balb/C 小鼠随机分为正常组、便秘组、益肠通秘汤组和莫沙必利组，每组10 只。除正常组外，其余组小鼠使用洛哌丁胺(10 mg/kg)灌胃 2 周建立便秘模型。第 3 周开始，益肠通秘汤组给予益肠通秘汤 14 g/(kgd)灌胃，莫沙必利组给予枸橼酸莫沙必利 5 g/(kgd)灌胃，正常组及便秘组给予等量生理盐水灌胃，均持续 2 周。灌胃结束后，观察各组小鼠粪便数量、粪便含水量及小肠活性炭推进长度、肠道推进率，使用 16S rDNA 测序方法分析各组小鼠粪便肠道菌群，使用免疫荧光染色方法检测小鼠结肠组织中 Occludin 表达情况。结果便秘组小鼠粪便数量、粪便含水量及小肠活性炭推进长度、肠道推进率均明显低于正常组(P 均 0 05);益肠通秘汤组和莫沙必利组小鼠粪便数量、粪便含水量、肠道推进率及益肠通秘汤组小肠活性炭推进长度均明显高于便秘组(P 均 0 05)。便秘组小鼠菌群多样性中 Chao1、shannon 水平和粪便中 Lactobacil-lus、Bifidobacterium、Prevotella、Akkermansia 相对丰度均明显低于正常组(P 均 0 05)，Coprococcus和 Clostridium 相对丰度均明显高于正常组(P 均 0 05);益肠通秘汤组 Chao1、shannon 水平和Lactobacillus、Bifidobacterium、Prevotella、Akkermansia 相对丰度均明显高于便秘组(P 均 0 05)，Coprococcus 和 Clostridium 相对丰度均明显低于便秘组(P 均 0 05);莫沙必利组各指标与便秘组及益肠通秘汤组比较差异均无统计学意义(P 均 0 05)。便秘组小鼠结肠上皮中 Occludin 表达较正常组少;益肠通秘汤组和莫沙必利组 Occludin 表达较便秘组多，但莫沙必利组略少于益肠通秘汤组。结论益肠通秘汤能够通过恢复肠道菌群的多样性，增加有益菌和减少有害菌，发挥维护肠道屏障的作用而改善便秘。关键词 便秘;肠道菌群;益肠通秘汤;Occludin;肠道屏障doi:10 3969/j issn 1008 8849 2022 22 006 中图分类号 332 文献标识码 A 文章编号 1008 8849(2022)22 3095 06 作者简介 黄玲，女，在读硕士研究生，研究方向为中医药治疗肛肠疾病。通信作者 林爱珍，E mail:Linaizhen2003163 com 基金项目 湖北省卫生健康委员会中医药科研项目(ZY2021M015)Effect of Yichang Tongmi decoction on diversity of bacterial flora and expressionof intestinal Occludin in constipated miceHUANG Ling1，PAN Jiao2，SHI Zhenyang1，LIN Aizhen3(1 Clinical College of Traditional Chinese Medicine，Hubei University of Traditional Chinese Medicine，Wuhan 430065，Hubei，China;2 Yueyang Peoples Hospital，Yueyang 414100，Hunan，China;3 Hubei Provincial Hospital of Tradition-al Chinese Medicine，Wuhan 430065，Hubei，China)Abstract:Objective It is to explore the effect of Yichang Tongmi decoction on the diversity of bacterial flora and theexpression of intestinal tight junction protein(Occludin)in mice with constipation Methods Forty Balb/C mice were ran-domly divided into normal group，constipation group，Yichang Tongmi decoction group and mosapride group，with 10 micein each group All the mice in each group except for the normal group were gavaged with loperamide(10 mg/kg)for 2weeks to establish constipation models From the third week，the Yichang Tongmi decoction group was given Yichang Tong-5903现代中西医结合杂志 Modern Journal of Integrated Traditional Chinese and Western Medicine 2022 Nov，31(22)mi decoction 14 g/(kg d)by gavage，the mosapride group was given mosapride citrate 5 g/(kg d)by gavage，thenormal group and the constipation group were given the same amount of normal saline by gavage，all groups were gavagedfor 2 weeks After gavage，the fecal quantity，fecal water content，active carbon advance length and intestinal advance rateof mice in each group were observed，the intestinal flora of mice in each group was analyzed by 16S rDNA sequencing meth-od，and the expression of Occludin in colon tissue of the mice was detected by immunofluorescence staining method e-sults The fecal quantity，fecal water content，intestinal active carbon propulsion length and intestinal propulsion rate ofmice in constipation group were significantly lower than those in normal group(all P 0 05)，the fecal quantity，fecal wa-ter content，intestinal propulsive rate in Yichang Tongmi decoction group and mosapride group and intestinal active carbonpropulsive length of mice in Yichang Tongmi decoction group were significantly higher than those in constipation group(allP 0 05)The levels of Chao1 and Shannon in the bacterial diversity and the relative abundance of Lactobacillus，Bifidobacterium，Prevotella，Akkermansia in feces of mice in constipation group were significantly lower than those in nor-mal group(all P 0 05)，while the relative abundance of Coprococcus and Clostridium were significantly higher than thosein normal group(all P 0 05);The levels of Chao1 and Shannon and the relative abundance of Lactobacillus，Bifidobac-terium，Prevotella，Akkermansia in Yichang Tongmi decoction group were significantly higher than those in constipationgroup(all P 0 05)，while the relative abundance of Coprococcus and Clostridium were significantly lower than those inconstipation group(all P 0 05);There was no significant difference in all indexes among mosapride group，constipationgroup and Yichang Tongbi decoction group(all P 0 05)The expression of Occludin in colonic epithelium of mice inconstipation group was less than that in normal group，the expression of Occludin in Yichang Tongmi decoction group andmosapride group was more than that in constipation group，but mosapride group was slightly less than Yichang Tongmi de-coction group Conclusion Yichang Tongmi decoction can improve constipation by restoring the diversity of intestinal flora，increasing beneficial bacteria and reducing harmful bacteria to maintain intestinal barrierKeywords:constipation;intestinal flora;Yichang Tongmi decoction;Occludin;intestinal barrier便秘主要以大便干结、排便次数减少和排便困难为主要表现，中老年人多发，不仅影响患者日常生活，严重者还可发展成结肠黑变病、结肠癌等1。现代研究已经证明便秘的发病受饮食、遗传、情绪的影响，同时与肠道菌群的