具核梭杆菌诱导的肠上皮细胞Caco-2炎症反应及凋亡  

Fusobacterium nucleatum induces inflammation and apoptosis in cultured intestinal epithelial Caco-2 cells

作　　者：王昆[1] 贾银平[1] 朱攀[1] 方瑶[1] 李倩[1] 唐彬[1,2] 李娜 毛旭虎[1] 

机构地区：[1]第三军医大学西南医院医学检验系临床微生物暨免疫学教研室,重庆400038 [2]成都军区峨眉疗养院,峨眉山614200 [3]济南军区总院心血管研究所,济南250031

出　　处：《免疫学杂志》2015年第4期313-317,共5页Immunological Journal

基　　金：全军“十二五”重大项目(AWS11J011-04)

摘　　要：目的研究具核梭杆菌诱导人肠上皮细胞(Caco-2)的炎症反应与凋亡,并初步探讨其机制。方法具核梭杆菌感染Caco-2细胞后,采用RT-PCR及ELISA检测促炎细胞因子IL-1β、IL-8和TNF-α的转录水平和蛋白水平的表达,利用透射电镜、流式细胞术、Western blot和RT-PCR等技术检测具核梭杆菌诱导的肠上皮细胞凋亡情况及促凋亡相关蛋白PARP剪切、BECN1、Caspase3剪切和AKT磷酸化的情况。结果具核梭杆菌感染人肠上皮细胞系Caco-2后,无论在转录水平还是蛋白水平,促炎细胞因子IL-1β、IL-8和TNF-α的表达均比对照组显著升高,其中感染6 h后促炎细胞因子表达最高。其次,具核梭杆菌能够诱导Caco-2细胞凋亡,其中感染6 h凋亡率最高;具核梭杆菌感染6 h后,凋亡通路相关蛋白PARP剪切、Caspase3剪切显著增加,BECN1显著上调,而抗凋亡蛋白AKT磷酸化则显著下调。结论具核梭杆菌能够诱导人肠上皮细胞的炎性反应和凋亡。To investigate the inflammation and apoptosis induced by Fusobacterium nucleatum（F. nucleatum）in human intestinal epithelial Caco-2 cells, we detected the expressions of IL-1β, IL-8 and TNF-α by RT-PCR and ELISA after intestinal epithelial cells Caco-2 infected with F. nucleatum. Furthermore, the expression levels of cleaved PARP, BECN1, caspase3, p-AKT and AKT were measured by TEM, flow cytometry, Western blot and RTPCR, respectively. The results showed that the m RNA and protein levels of pro-inflammatory cytokines as IL-1β,IL-8 and TNF-α were significantly increased in Caco-2 cells during F. nucleatum infection, and peaked at 6 hour post-infection. F. nucleatum also induced utmost apoptosis in Caco-2 cells at the same period. The expression levels of cleaved PARP, caspase3, and BECN1 were increased significantly, whereas anti-apoptosis p-AKT was down regulated after F. nucleatum infection. In conclusion, F. nucleatum can induce inflammatory response and apoptosis in intestinal epithelial Caco-2 cells.

关 键 词：具核梭杆菌 肠上皮细胞 炎症 凋亡 

分 类 号：R378.8[医药卫生—病原生物学]