Akkermansia muciniphila上清通过抑制内质网应激改善小肠黏膜损伤  

Akkermansia muciniphila supernatant improves intestinal mucosal injury by inhibiting endoplasmic reticulum stress

作　　者：张丽君 袁俊婕 赵经文 陈鑫[1,2] ZHANG Lijun;YUAN Junjie;ZHAO Jingwen;CHEN Xin(Department of Gastroenterology and Hepatology,General Hospital,Tianjin Medical University,Tianjin 300052,China;Tianjin Institute of Digestive Diseases,Tianjin Key Laboratory of Digestive Diseases,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院消化科,天津300052 [2]天津市消化疾病研究所,天津市消化病学重点实验室,天津300052

出　　处：《天津医科大学学报》2024年第6期528-534,共7页Journal of Tianjin Medical University

基　　金：天津市卫生健康科技项目(TJWJ2021MS005)。

摘　　要：目的:探究Akkermansia muciniphila(AKK)上清对非甾体类抗炎药(NSAIDs)导致小肠黏膜损伤的干预效果及作用机制。方法:将18只C57BL/6小鼠(6~8周龄)采用随机数字表法随机分为3组:正常对照组(CON组)、造模组(INDO组)、干预组(INDO+AKK组),每组6只。INDO组使用吲哚美辛构建NSAIDs小肠损伤模型,INDO+AKK组使用AKK上清灌胃。采集各组肠道标本,常规苏木精-伊红(HE)染色观察黏膜屏障形态,免疫组织化学染色法(IHC)观察黏膜屏障指标咬合蛋白(Occuldin)、闭锁小带蛋白(ZO1)、黏蛋白2(MUC2)的表达水平。采用实时荧光定量PCR(qRT-PCR)从mRNA水平验证MUC2、葡萄糖调节蛋白78(GRP78)、X盒结合蛋白1(XBP1s)、C/EBP坏死因子相关蛋白(CHOP)、激活转录因子4(ATF4)、激活转录因子6(ATF6)、真核翻译起始因子2α激酶3(PERK)等基因的表达变化。结果:小肠组织HE染色观察发现,与CON组相比,INDO组出现小肠黏膜结构破坏、绒毛明显减少等特征。INDO+AKK组小肠黏膜损伤减轻、绒毛丧失现象减少、黏膜及其下层的炎症细胞浸润区域也显著减小。IHC分析显示,与INDO组相比,INDO+AKK组Occuldin(F=11.48,P<0.05)、ZO1(F=68.10,P<0.001)、MUC2(F=19.93,P<0.01)表达水平增加。PAS结果显示,INDO+AKK组杯状细胞数量相对于INDO组明显增加(F=205.9,P<0.001)。qRTPCR结果发现,与INDO组相比,INDO+AKK组MUC2的基因(F=23.67,P<0.01)表达水平升高,GRP78(F=7.869,P<0.01)、CHOP(F=11.45,P<0.05)、XBP1s(F=8.344,P<0.05)、ATF4(F=16.37,P<0.001)、ATF6(F=12.99,P<0.001)、PERK(F=11.58,P<0.01)的基因表达水平均下降。结论:AKK上清通过抑制内质网应激,改善NSAIDs诱导的小肠黏膜屏障损伤。Objective:To investigate the effect and mechanism of Akkermansia muciniphila(AKK)supernatant on intestinal mucosal injury induced by non-steroidal anti-inflammatory drugs(NSAIDs).Methods:Eighteen C57BL/6 mice(6-8 weeks of age)were randomly divided into three groups by random number table method:normal control group(CON),modeling group(INDO)and intervention group(INDO+AKK),with 6 mice in each group.INDO group was treated with Indomethacin to establish the small intestine injury model of NSAIDs,INDO+AKK group was treated with AKK supernatant by oral gavage.Intestinal samples of each group were collected,and mucosal barrier morphology was observed by hematoxylin-eosin(HE)staining,mucosal barrier indexes occuldin,zona occlusa(ZO1),and mucin 2(MUC2)were observed by immunohistochemical staining(IHC),and goblet cell number was observed by periodic Schiff(PAS)staining.Real-time quantitative fluorescent PCR(qRT-PCR)was used to verify the expression changes of MUC2,glucose regulatory protein 78(GRP78),X-box binding protein 1(XBP1s),C/EBP necrosis factor related protein(CHOP),activated transcription factor 4(ATF4),activated transcription factor 6(ATF6),and eukaryotic translation initiation factor 2αkinase 3(PERK).and other genes at mRNA level.Results:HE staining of small intestine tissue revealed that compared with CON group,INDO group showed damage of intestinal mucosal structure and reduced villi.In the INDO+AKK group,the damage of small intestinal mucosa and the loss of villi were reduced,and the areas infiltrated by inflammatory cells in the mucosa and its sublayer were also significantly reduced.IHC analysis showed that Occuldin(F=11.48,P<0.05),ZO1(F=68.10,P<0.001)and MUC2(F=19.93,P<0.01)expression levels were increased in INDO+AKK group compared with INDO group.PAS results showed that the number of goblet cells in INDO+AKK group was significantly increased compared with INDO group(F=205.9,P<0.001).The results of qRT-PCR showed that compared with the INDO group,the expression level of MUC2 gene(F=23.67,P<0.01)was incr

关 键 词：AKK上清 内质网应激 NSAIDS 小肠黏膜损伤 

分 类 号：R574.5[医药卫生—消化系统]