【作者】 陈凝；

【导师】 彭孝纬；

【作者基本信息】 福建医科大学 ， 内科学， 2008， 硕士

【摘要】 目的:通过体外实验研究NF-κB信号通路在幽门螺杆菌引起慢性胃炎的机制中所起的作用,以及评价特异性调节NF-κB活性对于减轻幽门螺杆菌引起的慢性炎症反应的作用,为将来的动物及临床实验做好准备,为阻断慢性炎症向胃癌发展这一进程提供依据,从而为最终解决由幽门螺杆菌所引起的慢性胃炎等疾病提供有效的方案。方法:分别培养幽门螺杆菌11637标准菌株以及人胃粘膜上皮细胞(GES-1),将GES-1细胞分为3组:空白组、GES-1+Hp组及GES-1+Hp+NF-κB抑制剂组,每组各做10例,在后两组中按细胞与细菌为1:50的量加入Hp11637菌株,共培养24小时后,提取上清液用ELISA法检测IL-8、TNF-α的含量,裂解细胞,用Western Blot法检测NF-κB的表达含量。以空白组为对照分别比较Hp及NF-κB抑制剂对于NF-κB信号通路的影响作用。结果:1、成功培养Hp11637标准菌株及GES-1细胞。2、在GES-1细胞中加入Hp菌株后共培养24小时后提取上清液再次进行Hp培养,Hp培养成功。3、在GES-1细胞中按比例加入定量的Hp菌株,共培养24小时后检测上清液中的IL-8、TNF-α含量,较空白组明显升高(p<0.05),裂解细胞后检测裂解液中的NF-κB表达量较空白组升高(p<0.05)。4、在GES-1细胞中加入定量的Hp菌株以及特异性NF-κB信号通路抑制剂,共培养24小时后检测上清液中的IL-8、TNF-α含量,与仅加入Hp的干预组相比,表达量有所降低(p<0.05),裂解细胞后检测裂解液中的NF-κB表达量较干预组降低(p<0.05),但较空白组升高(p<0.05)。结论:NF-κB信号通路在幽门螺杆菌引起慢性胃炎的发病机制中起到核心作用,采用特异性阻断NF-κB信号通路的方法可以有效减少幽门螺杆菌诱导的炎性因子的分泌。更多还原

【Abstract】 Objective: Use in vitro experiment to study the function of NF-κB signal pathway in the pathogenic mechanism of chronic gastritis associated with helicobacter pylori infection to evaluate the effect of accommodation of NF-κB in abatement chronic inflammation associated with helicobacter pylori infection , to make up the basement of future animal or clinical experiment , so that we can stop the progress of chronic gastritis to gastric cancer and can support effect way to treat chronic gastritis associated with helicobacter pylori.Methods: Helicobacter pylori standard strain(11637 strain) and gastric epithelial cell were cultured. The GES-1 were divided in to three groups:the blank group, the GES-1 and Hp group, the GES-1,Hp and inhibitor of NF-κB group. Each group contains 10 examples. The GES-1 and Hp were put together at the ratio of 1:50 in the last two groups. After the three groups were cultured for 24 hours, the supernatant liquid was separated to detect the contents of IL-8 and TNF-αby ELISA. Then the cells were splitted to detect the content of NF-κB by Western Blot. The datas were compared between three groups to evaluate the influence of Hp and NF-κB inhibitor to the NF-κB signal pathway.Results: The Hp standard strain(11637 strain) and GES-1 were cultured successfully. 2.After GES-1 and Hp were cultured for 24 hours, the supernatant liquid was cultured in microaerobic atmosphere, then Hp can be cultured again. 3.After GES-1 and Hp were cultured together for 24 hours, the contents of IL-8 and TNF-αin the supernatant liquid were more than the blank group(p<0.05), and the content of NF-κB was more than the blank group(p<0.05). 4.After GES-1,Hp and NF-κB inhibitor were cultured for 24 hours, the contents of IL-8 and TNF-αin the supernatant liquid were less than the group of GES-1and Hp (p<0.05), the content of NF-κB was less than the group of GES-1 and Hp(p<0.05), but more than the blank group.Conclusions: The NF-κB signal pathway plays an important role in the pathogenic mechanism of chronic gastritis associated with helicobacter pylori infection. We can block the NF-κB signal pathway to decrease the secretion of inflammatal factor induced by helicobacter pylori.更多还原