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目的通过建立H_2O_2导致人脐静脉内皮细胞损伤的模型来研究狭鳕鱼皮胶原蛋白肽作用。方法用人脐静脉内皮细胞作为体外研究对象,采用不同的浓度(40、200、400μg/mL)胶原蛋白和维生素E预处理12h,然后在加入50μmol/L H_2O_2继续培养12h造成损伤模型。模型成功后用CCK-8检测吸光度,根据试剂盒操作检测细胞内及培养上清液谷胱甘肽过氧化物酶(GXH-PX),一氧化氮(NO)、过氧化氢酶(CAT),丙二醛(MDA)等指标的含量和活性变化。结果经胶原蛋白预保护后细胞增值率升高,GXH-PX、CAT等活性增强,NO含量升高,丙二醛含量降低。结论 50mmol/L H2O2能够成功复制人脐静脉内皮细胞损伤模型,胶原蛋白肽能够有效的对抗这种损伤作用且呈剂量依赖性。其保护作用机制可能与调控GXH-PX、CAT的活性以及NO、丙二醛的含量有关。
OBJECTIVE: To study the effect of collagen peptide of codfish skin by establishing a model of H 2 O 2 -induced injury of human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cells were used as in vitro study. Collagen and vitamin E at different concentrations (40, 200 and 400μg / mL) were pretreated with collagen and vitamin E for 12h, then cultured in HUVEC with 50μmol / L H2O2 for 12h. After the model was successfully used, the absorbance was measured by CCK-8. The activities of GXH-PX, NO and CAT in cells and culture supernatants were detected by kit operation. , Malondialdehyde (MDA) and other indicators of content and activity changes. Results After pre-protected by collagen, cell proliferation rate increased, GXH-PX, CAT activity increased, NO content increased, malondialdehyde content decreased. Conclusion 50 mmol / L H2O2 can successfully replicate human umbilical vein endothelial cell injury model. Collagen peptide can effectively resist this injury in a dose-dependent manner. The protective mechanism may be related to the regulation of GXH-PX, CAT activity and NO, MDA content.