摘要：
目的 探讨硫糖铝混悬凝胶(SC)对幽门螺杆菌(H.pylori)感染所致小鼠胃黏膜损伤和胃肠菌群的影响.方法 C57BL/6J小鼠分为正常对照组(NC组)、H.pylori组(HP组)和硫糖铝组(HP+SC组).HP组和HP+SC组以H.pylori悉尼菌株(SS1)灌胃,HP+SC组同时予SC灌胃14d.小鼠处死后取胃黏膜行苏木精-伊红(HE)染色并行胃黏膜损伤评分(EDS),免疫组织化学法(IHC)或改良吉姆萨染色阳性认为H.pylori阳性.IHC检测紧密连接蛋白及炎症因子并行免疫组化评分,实时荧光定量聚合酶链式反应(qPCR)检测炎症因子,透射电镜观察胃黏膜超微结构.取胃黏膜和肠内容物行16S核糖体RNA(16S rRNA)测序.结果 HP组小鼠H.pylori定植率为91.67％,HP+SC组定植率为50.00％.HP组小鼠胃黏膜EDS高于NC组[(3.42±0.19)比(1.17±0.17),P＜0.05],HP+SC组EDS(2.42±0.29)低于HP组(P＜0.05).HP组白细胞介素(IL)-8、IL-10、肿瘤坏死因子(TNF)-α mRNA表达水平及IHC评分高于NC组(均P＜0.05).HP+SC组IL-8 mRNA表达水平[(0.90±0.21)比(1.72±0.23)]及IHC评分[(4.50±0.22)分比(5.83±0.31)分]均低于HP组(均P＜0.05),而IL-10、TNF-α mRNA表达水平及IHC评分差异均无统计学意义(均P＞0.05).HP组闭锁小带蛋白(ZO-1)及咬合蛋白IHC评分低于NC组(均P＜0.05),HP+SC组咬合蛋白IHC评分高于HP组[(3.50±0.43)分比(2.33±0.21)分,P＜0.05],两组ZO-1的IHC评分差异无统计学意义(P＞0.05).HP组较NC组胃菌群多样性降低,肠道菌群多样性差异无统计学意义,HP+SC组较HP组胃菌群及肠道菌群差异无统计学意义.结论 SC降低H.pylori定植率,改善H.pylori所致胃黏膜损伤,降低H.pylori所致IL-8表达,增强咬合蛋白,但对H.pylori所致胃肠菌群改变无明显影响.%Objective To investigate the effects of sucralfate suspension gel (SC) on Helicobacter pylori(H.pylori)colonization,H.pylori-induced gastric mucosal injury and gastrointestinal microecology in mice.Methods C57BL/6J mice were randomly divided into 3 groups,including normal control (NC)group,H.pylori model (HP)group,and SC prevention (HP+SC)group.H.pylori infection mouse model was established by gavage with H.pylori Sydney strain (SS1).And HP+SC group was also administered with SC for 14 days.After mice were sacrificed,the gastric mucosa was taken for HE staining,immunohistochemical (IHC) staining [H.pylori,zonula occludens-1 (ZO-1),Occludin,interleukin (IL)-8,IL-10 and tumor necrosis factor-x (TNF-α)],qPCR(IL-8,IL-10,TNF-α).And cell ultrastructure was observed by electron microscopy.Microbiota communities in the gastric mucosa or fecal were investigated using 16S ribosomal RNA (rRNA) gene sequencing.The detection of H.pylori in IHC staining or Giemsa staining was defined as H.pylori infection.Results The H.pylori positive rate of HP group was 91.67％,and that of HP+ SC group was 50.00％.The EDS of HP group were significantly higher than those of the NC group [(3.42± 0.19) vs (1.17±0.17),P＜0.05],while EDS of HP+SC group (2.42±0.29) were significantly lower than that of HP group (P＜0.05).The mRNA levels and immunostaining scores of IL-8,IL-10 and TNF-α in HP group was higher than NC group (all P＜0.05).Compared with HP group,mRNA levels and IHC scores of IL-8 significantly decreased (all P＜0.05) in HP+SC group,while those of IL-10 and TNF-α was similar between two groups (all P＞0.05).The IHC scores of ZO-1 and Occludin in HP group was significantly lower than NC group (all P＜0.05),and that of ZO-1 and Occludin in the HP+SC group increased compared with HP group [(2.00±0.26) vs (1.17±0.48),P＞0.05;(3.50±0.43) vs (2.33±0.21),P＜0.05].Compared with NC group,the diversity of gastric microbiota in HP group was significantly lower,and diversity of gut microbiota decreased insignificantly.There was no significant difference between HP+SC group and HP group in gastric or gut microbiota.Conclusion SC reduces H.pylori colonization,protects H.pylori-induced gastric mucosal injury,decreases H.pylori-induced IL-8 expression,enhances Occludin.However,its effects on H.pylori-induced gastrointestinal microbiota disorders are limited.
