摘要
目的：观察不同类型幽门螺杆菌（Helicobacter pylori，Hpylori）对GES一1细胞间隙连接通讯功能（gap junctionin tercellular communication，GJIC）的影响，探讨与GJIc功能异常有关的Hpylori菌株类型。方法：将临床培养分离的不同日．pylori菌株类型包括东亚型CagA＋Hpylori、西方型CagA＋Hpylori及CagA—Hpylori与GES一1细胞共培养24，48h；对照组不加Mpylor培养24，48h。采用划痕标记荧光染料示踪技术（scrape—loading and dye transfer，SLDT）检测GES一1细胞GJIc功能，四甲基偶氮唑盐（methyl thiazolyl tetrazoli—Um bromide，MTT）比色法检测细胞增殖情况。结果：对照组培养24，48h后GES一1细胞荧光染料向邻近细胞最远传递4～5列，具有较强的GJIC功能；CagA—Hpylori组最远传递3列，GJIC功能减弱；西方型CagA＋H．pylori组最远传递l～2列，GJIC功能明显减弱；东亚型CagA＋Hpylori组荧光染料大多局限于划痕旁的单列细胞，仅极少数传递1～2列，GJIC功能明显减弱或缺失。CagA—Hpylori对细胞增殖无明显影响，西方型或东亚型CagA＋Hpylori在细菌／细胞比例为100：1和200：1时细胞增殖增强，但东亚型CagA＋Hpylori细菌／细胞比例400：1培养48h时细胞增殖受到抑制。结论：Hpylori抑制GES一1细胞GJIc功能，以CagA＋Hpylori特别是东亚型CagA＋H.pylori抑制作用更明显，Hpylori对GES一1细胞增殖的影响与毒力因子CagA有关。
Objective To determine the effect of different types of Helicobacter pylori （H. pylori） on the gap junction intercellular communication （GJIC） in GES-1 cells, and investigate the types of H. pylori related to the dysfunction of GJIC. Methods Different types of H. pylori clinical strains were isolated and cultured, including the East Asian CagA-positive H. pylori （ East Asian CagA ＋ H. pylori） , Western CagA-positive H. pylori （Western CagA ＋ H. pylori ） , and the CagA-negative H. pylori （CagA-H. pylori）. We co-cultured these H. pylori strains with GES-1 cells for 24 and 48 h, respectively. The control group was cultured without any H. pylori for 24 and 48 h. Change of the GJIC function in GES-1 cells was detected by the scrape-loading dye transfer （SLDT） technique. The cell proliferation of each group was examined by the methyl thiazolyl tetrazolium bromide （MTT） assay. Results The control group showed better GJIC function in the GES-1 cells, and the fluorescent dye migrated 4 -5 rows to the adjacent cells at 24 and 48 h. Compared with the control group, the GJIC function of GES-1 cells in the CagA - H. pylori group decreased and the fluorescent dye migrated 3 rows to the adjacent cells. Compared with the control group and the CagA - H. pylori group, the GJIC function of GES-1 cells in the Western CagA ＋ H. pylori group decreased and the fluorescent dye migrated 1 - 2 rows to the adjacent cells. The East Asian CagA ＋ H. pylori group showed no GJIC function or weak GJIC fimction, and most of the fluorescent dye was confined to the area of scratched single row cells and only a few migrated 1 -2 rows to the adjacent cells. Difference in the cell proliferation between the CagA-H. pylori group and the control group was not significant. The cell proliferation of the Western CagA ＋ H. pylori group and the East Asian CagA ＋ H. pylori group at bacterium-to-cell ratio of 100：1 and 200：1 was higher than that of the control group. The cell proliferation of the East Asian CagA ＋ H. pylori group at bacterium-to-cell ratio of 400 ： 1 was significantly lower than that of the control group at 48 h. Conclusion H. pylori can inhibit the GJIC function in GES-1 cells, which may be associated with CagA ＋H. pylori, especially with East Asian CagA ＋H. pylori. The effect of H. pylori on the proliferation of GES-1 cells is related to virulence factor CagA.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2011年第4期294-300,共7页
Journal of Central South University ：Medical Science
基金
supported by the Natural Science Foundation of Hunan Province,P.R.China(09JJ6047)
关键词
幽门螺杆菌
菌株类型
胃黏膜上皮细胞
细胞间隙连接通讯
细胞增殖
Helicobacter pylori
type of Helicobacter pylori strain
gastric epithelial cell
gap junction intercellular communication
cell proliferation
