江明礼,莫非,渠巍,孙朝琴,罗昭逊,张姝,吴银杰
JIANG Mingli,MO Fei,QU Wei,SUN Chaoqin,LUO Zhaoxun,ZHANG Shu,WU Yinjie

• 1:贵州医科大学
• 2:贵阳市儿童医院检验科

摘要(Abstract)：

目的:观察苗药头花蓼对感染幽门螺杆菌(H. pylori)胃炎细胞TLRs信号通路的影响。方法:将人永生化胃上皮细胞(GES-1)随机分为空白组、模型组及药物组,模型组和药物组H. pylori与GES-1细胞以100∶1的比例共培养12 h、空白组给予等量培养基,头花蓼组给予相应药物干预48 h、空白组及模型组给予等量培养基;干预结束后,采用光学及电子显微镜观察各组细胞形态结构的变化,并以Real-Time PCR及Western blot法检测各组GES-1细胞TLR2、TLR4、TLR5、TLR9及下游信号因子MyD88、TRAF6 mRNA和蛋白表达水平。结果:感染H. pylori后,模型组细胞镜下较空白组偏瘦长、表面粗糙、颗粒物质明显增多、细胞膜表面绒毛减少、伪足多肿胀,药物组细胞大体饱满、杂质颗粒明显减少、细胞膜边缘整齐、结构完整、伪足增多外泌体较少; Western blot及Real-time PCR结果显示,模型组GES-1细胞中TLR2、TLR4、TLR5、TLR9及MyD88、TRAF6 mRNA及蛋白表达较空白组明显升高(P <0. 05);与模型组比较,药物组GES-1细胞中TLR2、TLR4、TLR5、TLR9及MyD88、TRAF6mRNA及蛋白下调(P <0. 05)。结论:头花蓼可能通过抑制TLRs信号通路的激活,从而抑制下游炎症因子的激活和释放,改善H. pylori引起的相关炎症反应。
Objective: To observe the effect of Polygonum capitatum on TLRs signal pathway in Helicobacter pylori( H. Pylori) gastritis cells. Methods: The human immortalized gastric epithelial cells( GES-1) were randomly divided into blank group and experimental group. The experimental group included model group and Polygonum capitatum group. The experimental group was treated with H. pylori ATCC 700392,which was co-cultured with GES-1 cells for 12 h at a ratio of 100∶ 1 to establish a gastritis cell model infected with H. pylori. The blank group was given the same amount of medium. The Polygonum capitatum group was then given the corresponding drug intervention for 48 h and the blank group and the model group were given the same amount of medium. After the intervention,optical and electron microscope were used to observe morphological changes of each group,and Real-Time PCR and Western Blot were used to detect the expression of TLR2,TLR4,TLR5,TLR9 and downstream signaling molecules MyD88,TRAF6 mRNA and protein in GES-1 cells. Results: After treated with H. pylori,the cells of the model group were elongated,the surface was rough and the granular substance was obviously increased. The villi on the surface of the cell membrane were reduced compared with the blank group,and the pseudopod was swollen. The cells in the drug group were generally full and the impurity particles were obviously reduced. The cell membrane was obviously improved; the edges were neat; the structure was intact and the pseudopodia increased. The results of Western blot and Real time PCR showed that the expression of TLR2,TLR4,TLR5,TLR9,MyD88,TRAF6 mRNA and protein in GES-1 cells was significantly higher than that in the blank group( P < 0. 05); Compared with the model group,TLR2,TLR4,TLR5,TLR9 and MyD88,TRAF6 mRNA and protein were down-regulated in GES-1 cells( P < 0. 05). Conclusion: Polygonum capitatum may inhibit the activation of TLRs signal pathway,regulate the expression of innate immune receptors on cell membrane surface and thus improve the related inflammation induced by H. pylori.

关键词(KeyWords)： 苗药;头花蓼;螺杆菌,幽门;胃炎;Toll样受体;信号通路
Miao medicine;Polygonum capitatum;Helicobacter pylori;gastritis;Toll-like receptors;signal pathway

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技合作计划基金资助项目[黔科合LH(2015)7417号];; 贵州省卫生计生委科学技术基金资助项目(gzwjkj201 5-1-003);; 贵州省贵阳市科技计划项目[筑科合同(20141001)号];; 贵州省中医药管理局中医药、民族医药科学技术研究课题(QZYY-2016-005)

作者(Author): 江明礼,莫非,渠巍,孙朝琴,罗昭逊,张姝,吴银杰
JIANG Mingli,MO Fei,QU Wei,SUN Chaoqin,LUO Zhaoxun,ZHANG Shu,WU Yinjie

DOI: 10.19367/j.cnki.1000-2707.2019.02.012

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