奥美拉唑对大鼠反流性食管黏膜损伤后细胞增殖相关基因表达的影响

Effects of omeprazole on the esophageal mucosal cell proliferation and expressions of related genes in rats

目的观察奥美拉唑对反流性管黏膜损伤后细胞增殖相关基因表达的影响。方法健康雄性SD大鼠随机分为5组,假手术组、单纯胃食管反流动物模型组(模型组)、模型组+奥美拉唑低剂量组、中剂量组、高剂量组,每组15只。分别取食管标本肉眼观察黏膜状况,HE染色,光镜观察组织病理学变化,应用免疫组化染色等方法检测p21,PCNA,CyclinD1,CDK4的表达情况。结果假手术组大鼠食管颜色淡红,有光泽,黏膜光滑。模型组大鼠食管颜色变淡白,黏膜粗糙、增厚、皱襞灰白色斑块状增生。与模型组比较,奥美拉唑低、中、高剂量治疗组大鼠食管病变明显减轻(P<0.05)。HE染色显示,假手术组大鼠食管各层结构完整,黏膜厚薄适中,黏膜基底膜清楚,各层细胞排列整齐,表面可见较少的角化。模型组大鼠黏膜层增厚,基底细胞层及棘细胞层细胞数目增多,角化层增厚,局部黏膜基底细胞层明显增生,部分黏膜组织向外呈乳头样突起。与模型组比较,奥美拉唑治疗组大鼠食管黏膜损伤及增生病变明显减轻(P<0.05)。免疫组织化学检测显示,假手术组大鼠食管黏膜有p21蛋白表达,PCNA,CyclinD1和CDK4蛋白呈阳性基础表达。模型组大鼠食管黏膜p21蛋白的表达减弱,PCNA,CyclinD1和CDK4蛋白表达显著增强(P<0.05)。奥美拉唑治疗组大鼠食管黏膜p21蛋白的表达增强,PCNA,CyclinD1和CDK4蛋白表达明显降低(P<0.05)。结论奥美拉唑抑制细胞增殖可能是其治疗反流性食管损伤的重要机制。

Objective To observe the effects of omeprazole on esophageal mucosal cell proliferation and expressions of related genes in rats with esophageal mucosal injury induced by reflux of acid.Methods Seventy-five healthy male SD rats,weighted 220~250g,were randomly divided into 5 groups： sham-operation group,model group （ simple gastroesophageal reflux model） ,omeprazole low-dose group,medium-dose group and high-dose group,with 15 rats in each group.The rat models of reflux esophagitis were induced by single acid reflux.The pathological changes of esophageal mucosal were examined by naked eye, HE staining and light microscopy.The expressions of p21, PCNA, CyclinD1 and CDK4 were detected by immunohistochemical staining,and the results were analyzed by HPIAS-1000 system.The data were statistically analyzed with SPSS10.0.Results The color of esophagus of rats was erythroid and lustrous, with glossy mucosal membranes in sham-operation group.As compared with that in sham-operation group,the esophagus presented light white and esophageal mucous membrane became rough, thickened, grey plaque-like proliferation in rats of model group.As compared with that in model group,the pathological changes of esophagus were significantly relieved in the three different doses omeprazole treatment groups （ P 〈0.05） .HE staining showed that the various layer structure of rat’ s esophagus was intact,the base membrane of mucosa was clear and cellular arrangement was orderly in sham-operation group.As compared with those in sham-operation group,the esophageal mucosa was thickened,the cell numbers were increased in basal cell layer and prickle cell layer,and the cuticulae layer was thickened and proliferated,with some mucosal tissue presenting nipple-like elevation in model group.As compared with those in model group,the pathological changes of esophageal mucosa were significantly alleviated in omeprazole treatment groups （ P 〈0.05 ） .The results by immunohistochemistry showed that there was positive expression of p21 and basic expression of PCNA, CyclinD1 and CDK4 in basal cell layer of esophageal mucous membrane in sham-operation group. However the expression of p21 was decreased,the expressions of PCNA,CyclinD1,CDK4 were significantly increased in model group （ P 〈0.05）.As compared with those in model group,the expression of p21 was increased,and the expressions of PCNA,CyclinD1,CDK4 were significantly decreased in omeprazole treatment groups （ P 〈0.05）.Conclusion Omeprazole can up-regulate the expression of p21 after reflux esophagus injury and down-regulate the expressions of PCNA,CyclinD1 and CDK4,moreover,its inhibition effect on cell proliferation may be important action mechanism in treatment of reflux esophagus injury.