幽门螺杆菌感染小鼠胃黏膜组织细胞因子变化的实验研究

目的：研究幽门螺旋杆菌（简称幽门螺杆菌）感染小鼠胃黏膜组织细胞因子变化，初步探讨幽门螺杆菌感染的免疫机制。方法选取8～10周龄BALB／c小鼠24只，雌雄各半，分为观察组和对照组，两组造模前均禁食12h，观察组以2×109CFU／mL浓度的幽门螺杆菌菌液0．5mL／d灌胃，连续7d。对照组灌胃给生理盐水，0．5mL／d，连续7d，进行幽门螺杆菌感染小鼠胃黏膜造模。采用实时定量荧光PCR（Real‐timePCR）检测检测感染2、4周后，小鼠血浆中γ干扰素（IFN‐γ）、重组改构人肿瘤坏死因子α（TNF‐α）、白细胞介素‐8（IL‐8）以及IL‐10mRNA的表达量，并于感染4周后，处死小鼠，无菌取各组小鼠的胃组织，采用酶联免疫吸附试验夹心法检测胃黏膜组织中IFN‐γ、TNF‐α、IL‐8及IL‐10蛋白的水平。结果两组小鼠感染2周后，观察组IL‐8高于对照组，差异有统计学意义（P＜0．05）。感染4周后，IFN‐γ、TNF‐α、IL‐8的mRNA及蛋白表达量均明显上升，但观察组上升幅度明显高于对照组，差异有统计学意义（P＜0．05）。结论TH2细胞在幽门螺杆菌感染中的作用表现不明显，能促进TH1型细胞细胞因子的表达，引发TH1为主的免疫反应。

Experimental study of cytokines in gastric mucosa of mice with Helicobacter pylori infection

Objective To study the change of cytokines in gastric mucosa of mice with Helicobacter pylori infection and explore the immune mechanism .Methods Twenty‐four 8-10 weeks‐old BALB/c mice were randomly divided into observation group and control group .After first 12 hours of fasting ,observation group was given 0 .5 mL/d Helicobacter pylori bacterial liquid with con‐centration of 2 × 109 CFU/mL for 7 consecutive days .The control group was fed with 0 .5 mL/d saline for 7 consecutive days .Then the mice were infected with Helicobacter pylori gast model was established .Using Real‐time PCR to detect the expressions of IFN‐γ ,TNF‐α,IL‐8 and IL‐10 in murine plasma within 2 ,4 weeks after infection .And after 4 weeks of infection ,mice were sacrificed , and their stomach tissue sterile were separated .ELISA was used to detect the levels of IFN‐γ,TNF‐α,IL‐8 ,and IL‐10 mRNA in gastric mucosa .Results There was significantly difference between groups 2 weeks after infection(P<0 .05) .With lastingness of infection ,the expression and the mRNA of IFN‐γ,TNF‐α,IL‐8 were significantly increased ,and the rate of increase of observation group was higher(P<0 .05) .Conclusion TH2 cells may have no effect on Helicobacter pylori infection in mice gastric mucosa and promote the expression of T H1‐type cytokines ,triggering immune response of T H1 .