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目的探讨重组幽门螺杆菌过氧化氢酶(rHpCAT)对大鼠结肠粘膜上皮细胞氧化应激的影响。方法在分离培养正常大鼠结肠粘膜上皮细胞的基础上,利用FeSO4+H2O2反应产生羟自由基攻击制备结肠粘膜细胞氧化损伤模型,同时预先加入rHpCAT并观察损伤减轻程度。实验设正常对照组、模型对照组、5-氨基水杨酸给药组(0.1 mmol/L)、重组幽门螺杆菌过氧化氢酶给药组(1×105、1×106、1×107 U/kg·b.w.)。培养一定时间后,取上清测乳酸脱氢酶(LDH)、丙二醛(MDA) 和髓过氧化物酶(MPO)的活性进行测定,并同时测定上述各组谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)和超氧化物歧化酶(SOD)含量。结果模型组大鼠结肠粘膜上皮细胞MDA、LDH和MPO水平增高,CAT、GSH-Px和SOD含量减少。不同单位rHpCAT呈剂量依赖性的减少LDH释放,抑制MDA和MPO的形成,而使CAT、GSH-Px及SOD恢复到正常水平。结论重组幽门螺杆菌过氧化氢酶对大鼠结肠粘膜氧化损伤具有保护作用。
Objective To investigate the effect of recombinant H. pylori catalase (rHpCAT) on oxidative stress in colonic epithelial cells of rats. Methods The colonic mucosal epithelial cells of normal rats were isolated and cultured. The oxidative damage model of colonic mucosal cells was established by FeSO 4 + H2O2 reaction to produce oxidative damage of colonic mucosa. At the same time, rHpCAT was added to observe the extent of lesion. In the experiment, normal control group, model control group, 5-aminosalicylic acid administration group (0.1 mmol / L), recombinant H. pylori catalase administration group (1 × 105, 1 × 106, 1 × 107 U / kg · bw). After cultured for a certain period of time, the activities of lactate dehydrogenase (LDH), malondialdehyde (MDA) and myeloperoxidase (MPO) were measured and the glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) content. Results The levels of MDA, LDH, MPO and the contents of CAT, GSH-Px and SOD in colonic mucosal epithelial cells of model rats decreased. Different units of rHpCAT in a dose-dependent manner to reduce the release of LDH, inhibit the formation of MDA and MPO, leaving CAT, GSH-Px and SOD returned to normal levels. Conclusion Recombinant H.pylori catalase has a protective effect on oxidative damage of colonic mucosa in rats.