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目的探讨石榴皮多酚对大鼠应激性胃溃疡的保护作用及与Gli1-Gli2-Sufu信号通路和细胞因子水平改变的关系。方法选取50只7周龄健康SD雄性大鼠为受试对象,按随机数字表法分为空白对照组、应激性胃溃疡组(模型组)、奥美拉唑+应激性胃溃疡模型组(阳性对照组)、石榴皮多酚组和应激性胃溃疡模型+石榴皮多酚组,每组10只。分别比较各组大鼠胃液pH值和黏膜损伤指数;采用ELISA法测定前列腺素E2(PGE2)、促肾上腺皮质激素(ATCH)和皮质酮(CORT)的含量,应用化学比色法检测丙二醛(MDA)和超氧化物歧化酶(SOD);采用qPCR和Western blot法测定各组大鼠胃组织Gli1、Gli2和Sufu的mRNA和蛋白表达水平;免疫共沉淀法观察大鼠胃组织Gli2和Sufu蛋白复合物的形成情况。结果与空白对照组比较,模型组大鼠的损伤指数、MDA、ACTH和CORT的含量明显升高,pH值、SOD和PGE2水平显著下降,差异均有统计学意义(P<0.05);但阳性对照组和石榴皮多酚+模型组的上述各项指标均较模型组明显改善(P<0.05);模型组大鼠胃组织的Gli1、Gli2和Sufu的mRNA和蛋白表达均较空白对照组明显升高(P<0.05);而阳性对照组和石榴皮多酚+模型组的Gli1、Gli2和Sufu的mRNA和蛋白表达水平较模型组却明显降低(P<0.05);模型组大鼠Gli2和Sufu蛋白复合物水平较空白对照组显著增多,而阳性对照组和石榴皮多酚+模型组的Gli2和Sufu蛋白复合物水平则较模型组明显降低(P<0.05)。结论石榴皮多酚对大鼠应激性胃溃疡的保护作用可能与Gli1-Gli2-Sufu信号通路活化和MDA、ACTH、CORT、SOD和PGE2等细胞因子水平改变有关。
Objective To investigate the protective effect of pomegranate peel polyphenols on stress ulcer in rats and its relationship with Gli1-Gli2-Sufu signaling pathway and cytokine levels. Methods Fifty healthy 7-week-old male Sprague-Dawley rats were randomly divided into blank control group, stress gastric ulcer group (model group), omeprazole + stress ulcer model Group (positive control group), pomegranate peel polyphenol group and stress ulcer model + pomegranate peel polyphenol group, with 10 rats in each group. The gastric juice pH value and mucosal injury index of rats in each group were compared respectively. The levels of prostaglandin E2 (PGE2), adrenocorticotropic hormone (ATCH) and corticosterone (CORT) were determined by ELISA. The contents of malondialdehyde (MDA) and superoxide dismutase (SOD). The mRNA and protein expressions of Gli1, Gli2 and Sufu in gastric tissues were determined by qPCR and Western blot. The expressions of Gli2 and Sufu mRNA in gastric tissues were detected by co-immunoprecipitation Protein complex formation. Results Compared with the blank control group, the injury index, the content of MDA, ACTH and CORT in model group were significantly increased, while the levels of pH, SOD and PGE2 were significantly decreased (P <0.05), but the positive The contents of Gli1, Gli2 and Sufu in the model group were significantly higher than those in the blank control group (P <0.05) (P <0.05). However, the mRNA and protein expressions of Gli1, Gli2 and Sufu in the positive control group and the pomegranate polyphenol + model group were significantly lower than those in the model group (P <0.05). The levels of Gli2 and Sufu protein complex levels increased significantly compared with the blank control group, while the Gli2 and Sufu protein complex levels of the positive control group and the pomegranate polyphenol + model group was significantly lower than the model group (P <0.05). Conclusion The protective effect of pomegranate peel polyphenols on gastric ulcer in rats may be related to the activation of Gli1-Gli2-Sufu signaling pathway and the changes of cytokines such as MDA, ACTH, CORT, SOD and PGE2.