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目的建立幽门螺杆菌(Hp)感染蒙古沙鼠模型,探讨线粒体途径在Hp诱导胃上皮细胞凋亡中的作用。方法 48只雄性蒙古沙鼠均分为Hp感染组和对照组,每组分别于1、3和6个月3个时相点各处死8只动物,取胃黏膜行组织学检查:用Warthin-Starry银染、PCR和快速尿素酶法检测 Hp;通过H-E染色,光镜下观察胃黏膜病理变化;流式细胞仪测定细胞凋亡、线粒体膜电位及胞内游离 Ca2+含量。结果 Hp感染蒙古沙鼠后,胃黏膜出现慢性胃炎、肠化生及异型增生改变,而对照组胃黏膜基本正常,Hp感染组肠化生及异型增生发生率明显高于对照组(P<0．05)。Hp感染胃上皮细胞1、3、 6个月后的凋亡率分别为(16．71±3．30)％、(5．90±0．82)％、(5．69±0．70)％,而对照组的凋亡率分别为(4．20±0．94)％、(3．17±0．43)％、(4．70±0．55)％。其中Hp感染1个月后胃上皮细胞的凋亡率高于其他各组(P<0．05)。Hp感染胃上皮细胞1、3、6个月后的线粒体膜电位分别为43．10±17．62、 71．19±38．03、80．56±32．90,而对照组分别为84．70±23．50、84．39±37．51、79．54±30．24,其中Hp 感染1个月后胃上皮细胞的线粒体膜电位低于其他各组(P<0．05);Hp感染1、3、6个月后胃上皮细胞内游离Ca2+含量分别为18．60±9．32、5．18±2．06、4．94±3．25,而对照组分别为4．82±3．70、6．86 ±2．34、5．28±3．13,Hp感染1个月后胃上皮细胞内游离Ca2+含量高于其他各组(P<0．05)。结论 Hp诱导蒙古沙鼠胃上皮细胞凋亡主要发生在Hp感染早期;线粒体膜电位的下降和胞内游离Ca2+含量的升高参与了Hp诱发蒙古沙鼠胃上皮细胞凋亡的过程。
Objective To establish H. pylori infection model of Helicobacter pylori (Hp) in rats and explore the role of mitochondrial pathway in Hp-induced gastric epithelial cell apoptosis. Methods 48 Mongolian gerbils were divided into Hp infection group and control group. Each group was sacrificed at 3, 6 and 6 months respectively. Eight animals were sacrificed at each time point. Gastric mucosa were examined histologically with Warthin- Starry silver staining, PCR and rapid urease assay were used to detect Hp. The pathological changes of gastric mucosa were observed under light microscope by HE staining. Apoptosis, mitochondrial membrane potential and intracellular free Ca2 + content were determined by flow cytometry. Results The gastric mucosa showed chronic gastritis, intestinal metaplasia and dysplasia in H. pylori infected with H. pylori, while the gastric mucosa in control group was almost normal. The incidence of intestinal metaplasia and dysplasia in H. pylori infection group was significantly higher than that in control group (P <0. .05). The apoptotic rate of Hp-infected gastric epithelial cells at 1, 3 and 6 months were (16.71 ± 3.30)%, (5.90 ± 0.82)% and (5.69 ± 0.70) %, While the apoptotic rates in the control group were (4.20 ± 0.94)%, (3.17 ± 0.43)%, (4.70 ± 0.55)%, respectively. The apoptosis rate of gastric epithelial cells after Hp infection for 1 month was higher than other groups (P <0.05). Mitochondrial membrane potential of Hp-infected gastric epithelial cells at 1, 3 and 6 months were 43.10 ± 17.62, 71.19 ± 38.03 and 80.56 ± 32.90, respectively, while the control group was 84. 70 ± 23.50, 84.39 ± 37.51 and 79.54 ± 30.24, respectively. The mitochondrial membrane potential of gastric epithelial cells in Hp infected group was lower than that in other groups (P <0.05) In 1,3,6 months after infection, the content of intracellular free Ca2 + in gastric epithelial cells were 18.60 ± 9.32, 5.18 ± 2.06 and 4.94 ± 3.25, respectively, while the control group were 4.82 ± 3.70, 6.86 ± 2.34 and 5.28 ± 3.13, respectively. The content of intracellular free Ca2 + in gastric epithelial cells one month after Hp infection was higher than other groups (P <0.05). Conclusions The apoptosis of gastric epithelial cells in Mongolian gerbils mainly occurs in the early stage of Hp infection induced by Hp. The decrease of mitochondrial membrane potential and the increase of intracellular free Ca2 + are involved in the process of Hp-induced apoptosis of gastric epithelial cells in Mongolian gerbils.