Akkermansia muciniphila在结肠炎相关结肠癌中的作用和机制  

Role and mechanism of Akkermansia muciniphila incolon cancer associated with colitis

作　　者：刘益娟 魏凯艳 胡惠祥 王承党[1,2,3] 张琳[3,4] LIU Yijuan;WEI Kaiyan;HU Huixiang;WANG Chengdang;ZHANG Lin(Department of Gastroenterology,National Regional Medical Center,Binhai Campus of the First Affiliated Hospital,Fujian Medical University,Fuzhou,Fujian 350212,China)

机构地区：[1]福建医科大学附属第一医院滨海院区国家区域医疗中心消化内科,福建福州350212 [2]福建医科大学附属第一医院消化内科 [3]福建医科大学第一临床医学院 [4]福建医科大学附属第一医院胃肠外科

出　　处：《中国微生态学杂志》2023年第8期900-907,916,共9页Chinese Journal of Microecology

基　　金：福建省科技厅创双高联合基金(2019Y9120)。

摘　　要：目的 探究Akkermansia muciniphila在结肠炎相关结肠癌中的作用以及Akkermansia muciniphila通过影响肠道黏膜屏障延缓结肠炎癌变的机制。方法 C57BL/C小鼠共30只,分为对照组、结肠炎相关结肠癌组、结肠炎相关结肠癌+AKK组,每组10只,在建模结束后采用断头法处死小鼠,HE染色检测结肠病理结构,计算成瘤率。采用ELISA法检测3组小鼠结肠组织IL-1β、IL-6、TNF-α水平,免疫荧光双染法检测3组小鼠ZO-1和E-cadherin的表达情况,免疫组化法检测3组小鼠结肠组织Occludin-1、 N-cadherin、GPCR41、GPCR43、Bcl-2、 Bax等蛋白的水平,GC-MS法检测3组小鼠短链脂肪酸的水平。结果 HE发现结肠炎相关结肠癌组小鼠的结肠组织炎症浸润,肠腺水肿,存在不同程度的异型增生,结肠炎相关结肠癌+AKK组小鼠的结肠水肿和出血减轻,肠腺结构清晰。结肠炎相关结肠癌组中小鼠IL-1β、IL-6、TNF-α水平(pg/mL)(9.12±0.36、 6.36±0.42、 45.20±3.63)较对照组(3.80±0.38、 0.06±0.89、 2.10±0.14)升高(Tamhane’s T2分别为0.001、2.549、12.831,均P<0.05),经Akkermansia muciniphila灌胃后,IL-1β、IL-6、TNF-α水平(3.92±0.19、3.54±1.28、18.20±8.47)下降(Tamhane’s T2分别为6.553、2.414、5.743,均P<0.05)。免疫荧光双染法显示ZO-1和E-cadherin主要定位在结肠柱状上皮和肠腺,结肠炎相关结肠癌组ZO-1和Ecadherin的表达(ng/mL)(0.04±0.01、0.34±0.55)比对照组(0.10±0.18、0.48±0.13)降低(Tamhane’s T2分别为0.681、4.379,均P<0.05),而经Akkermansia muciniphila处理后显著上调小鼠结肠的ZO-1和E-cadherin的表达(0.08±0.05、1.08±0.28)(Tamhane’s T2分别为7.059、5.873,均P<0.05)。免疫组化显示结肠炎相关结肠癌组Occludin-1、 GPCR41、 GPCR43、 Bcl-2表达(ng/mL)(0.36±0.06、 0.48±0.13、 0.38±0.13、 0.34±0.55)较对照组(1.08±0.08、 0.74±0.06、 0.48±0.13、 1.64±0.11)减低(LSD-t分别为2.369、 0.304、 8.119、 2.298,均P<0.05),Bax表达(ng/mL)(1.34±0.27)较�Objective To explore the role of Akkermansia muciniphila(A.muciniphila)in colitis-related colon cancer(CC)and the mechanism of its effect on intestinal mucosal barrier in delaying colitis cancerosis.Methods A total of 30 C57BL/C mice were divided into blank control group,colitis associated CC group and colitis associated CC+AKK group,with 10 mice in each group.After modeling,the mice were sacrificed with decapitated method.HE was used to detect the pathological structure of colon,and the tumor formation rate was calculated.The levels of IL-1β,IL-6 and TNF-αin colon tissues in the 3 groups were detected with ELISA,and the expressions of ZO-1 and E-cadherin in the 3 groups were detected with immunofluorescence double staining.The protein levels of Occludin-1,N-cadherin,GPCR41,GPCR43,Bcl-2 and Bax in the colon tissue in the 3 groups were detected using immunohistochemistry,and the levels of short-chain fatty acids in the 3 groups were detected with GC-MS.Results HE found that colitis related CC group had inflammatory infiltration of colon tissue,intestinal glandular edema,and different degrees of dysplasia.Colitis related CC+AKK group had reduced colonic edema and bleeding,and intestinal glandular structure was clear.The contents of IL-1β,IL-6 and TNF-αin colitis associated CC group(pg/mL)(9.12±0.36,6.36±0.42,45.20±3.63,respectively)were higher than those in blank control group(3.80±0.38,0.06±0.89,2.10±0.14,respectively)(Tamhane's T2 were:0.001,2.549,12.831,all P<0.05),and those of IL-1β,IL-6 and TNF-αdecreased after inoculated with A.muciniphila(respectively:3.92±0.19,3.54±1.28,18.20±8.47)(Tamhane's T2 was 6.553,2.414,5.743,all P<0.05).Immunofluorescence double staining showed that ZO-1 and E-cadherin were mainly located in colon columnar epithelium and intestinal gland.The expressions of ZO-1 and E-cadherin in colitis related CC group(ng/mL)(0.04±0.01,0.34±0.55,respectively)were lower than those in blank control group(0.10±0.18,0.48±0.13)(Tamhane's T2 was:0.681,4.379,all P<0.05),and A.muciniphil

关 键 词：Akkermansia muciniphila 结肠炎 结肠癌 黏膜屏障 炎症因子 

分 类 号：R574.1[医药卫生—消化系统]