肠源反应性关节炎中HLA-B27表达情况及其与肠道细菌相互作用机制研究  

HLA-B27 expression in intestinal reactive arthritis and its interaction with intestinal bacteria

作　　者：李路[1] 王进海[1] 时志斌[2] 秦斌[1] 沙素梅 史海涛[1] LI Lu;WANG Jinhai;SHI Zhibin;QIN Bin;SHA Sumei;SHI Haitao(Department of Gastroenterology,the Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China;Department of Orthopedics,the Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China)

机构地区：[1]西安交通大学第二附属医院消化内科,陕西西安710004 [2]西安交通大学第二附属医院骨科,陕西西安710004

出　　处：《胃肠病学和肝病学杂志》2021年第1期102-108,共7页Chinese Journal of Gastroenterology and Hepatology

基　　金：陕西省重点研发计划项目(2017SF-100)。

摘　　要：目的探讨HLA-B27与反应性关节炎患者肠道沙门氏菌(S.enteritidis)胞内清除、复制、运输的机制情况。方法采用共聚焦显微镜、流式细胞术、夹心酶联免疫吸附试验(ELISA)评估相关蛋白的定位,评估肠杆菌在细胞内的转运,并测量相关细胞因子的产生。结果通过基因检测后发现,与对照组比较,病例组HLA-B27表达明显升高,差异有统计学意义(P<0.05);HLA-B27+/ReA+处理的细胞中细菌数量多于HLA-B27-/ReA-细胞。与HLA-B27-/ReA-细胞相比,HLA-B27+/ReA+细胞清除细胞内细菌较少(分别在4 h、24 h、72 h时,P<0.05);在感染后24 h、72 h、7 d,HLA-B27细胞中含有肠炎的细胞比HLA-A2细胞中明显增多,差异有统计学意义(P<0.05);通过ELISA法检测肠炎感染后不同组的TNF-α和IL-10的表达情况发现,与HLA-B27-/ReA-细胞比较,HLA-B27+/ReA+细胞TNF-α的含量明显升高,差异有统计学意义(P<0.05);而IL-10的表达情况同样在24 h后明显下降,差异有统计学意义(P<0.05);在HLA-B27细胞和HLA-A2细胞中,含有吞噬体的肠内菌体均可表达LAMP1和LAMP2。感染后30 min~24 h内,60%以上的含有GFP-S的吞噬体中检测到LAMP1和LAMP2;在感染后30 min和24 h,在HLA-B27细胞和HLA-A2细胞中仅检测到12%~17%的含有肠杆菌素的液泡中的转铁蛋白。低于20%的HLA-B27阳性转染细胞与30 min和24 h的对照细胞中发现了含有细菌的吞噬体和内吞标志物。结论反应性关节炎患者中HLA-B27呈阳性且其可能通过影响肠道细菌的存活及转运进行影响疾病的发生、发展。Objective To investigate the correlation between HLA-B27 and intracellular clearance,replication,and transport of S.enteritidis in patients with reactive arthritis.Methods In this study,confocal microscopy,flow cytometry,and sandwich enzyme-linked immunosorbent assay(ELISA)were used to evaluate the localization of related proteins,assess the transport of enterobacteria in cells,and measure the production of related cytokines.Results After genetic testing,compared with the control group,the expression of HLA-B27 in the case group was significantly increased and the difference was statistically significant(P<0.05);the number of bacteria in HLA-B27+/ReA+treated cells was more than that in HLA-B27-/ReA-cells.Compared with HLA-B27-/ReA-cells,HLA-B27+/ReA+cells eliminated fewer intracellular bacteria(P<0.05 at 4 hours,24 hours,and 72 hours,respectively);24 hours,72 hours,and 7 days after infection,HLA-B27 cells had significantly more enteritis-containing cells than HLA-A2 cells,and the difference was statistically significant(P<0.05);the expression of TNF-αand IL-10 in different groups after enteritis infection was detected by ELISA.Compared with HLA-B27-/ReA-cells,the content of TNF-αin HLA-B27+/ReA+cells increased significantly and the difference was statistically significant(P<0.05);the expression of IL-10 also appeared after 24 hours,significantly decreased and the difference was statistically significant(P<0.05);in HLA-B27 cells and HLA-A2 cells,both intestinal bacteria containing phagosomes could express LAMP1 and LAMP2.LAMP1 and LAMP2 were detected in more than 60%of phagosomes containing GFP-S within 30 minutes to 24 hours after infection.At 30 minutes and 24 hours after infection,only 12%-17%of transferrins in enterobactin-containing vacuoles were detected in HLA-B27 cells and HLA-A2 cells.Bacterial phagosomes and endocytic markers were found in less than 20%of HLA-B27 positive transfected cells and in control cells at 30 minutes and 24 hours.Conclusion HLA-B27 is positive in patients with reactive arthritis

关 键 词：反应性关节炎 HLA-B27 肠道细菌 相互作用 

分 类 号：R57[医药卫生—消化系统] R684.3[医药卫生—内科学]