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目的探讨狭鳕鱼皮胶原蛋白肽对H_2O_2诱导Ha Ca T细胞氧化损伤保护作用及分子作用机制。方法狭鳕鱼皮胶原蛋白肽50,100,200 mg·L~(-1)组预处理12 h后,加入300μmol·L~(-1)的H_2O_2培养12 h。采用酶生化法测定细胞上清液中过氧化氢酶(CAT),谷胱甘肽过氧化物酶(GSH-Px),总超氧化物歧化酶(T-SOD)和总抗氧化能力(T-AOC)的水平。Western蛋白印迹法检测细胞内水通道蛋白质3(AQP3)表达水平。结果与正常组相比,狭鳕鱼皮胶原蛋白肽浓度小于300 mg·L~(-1)时无细胞毒性。H_2O_2300μmol·L~(-1)组的细胞存活率降低到54.0%(P<0.01)。预先给狭鳕鱼皮胶原蛋白肽50,100,200 mg·L~(-1)组处理12 h使得细胞存活率相应增加到69.4%,79.4%和89.1%(P<0.05,P<0.01)。与模型组相比,狭鳕鱼皮胶原蛋白肽50,100,200 mg·L~(-1)组的CAT,GSH-Px,T-SOD和T-AOC含量提高,其中200 mg·L~(-1)组的CAT,GSH-Px,T-SOD和T-AOC含量相应升高了56.90%,48.68%,48.24%和71.43%(P<0.01)。与模型组相比,狭鳕鱼皮胶原蛋白肽200 mg·L~(-1)组AQP3表达量减少得最多,降低到67.1%(P<0.01)。结论狭鳕鱼皮胶原蛋白肽可保护H_2O_2诱导的氧化损伤,其机制可能与增强抗氧化能力和减少AQP3表达水平有关。
Objective To investigate the protective effect and the molecular mechanism of Polypeptide Cephalotaxus on the oxidative injury induced by H_2O_2 in HaCa T cells. Methods After pretreated with 50, 100, 200 mg · L -1 collagen peptide of Cape codfish skin for 12 h, 300 μmol·L -1 H 2 O 2 was added to culture for 12 h. The activities of catalase (CAT), glutathione peroxidase (GSH-Px), total superoxide dismutase (T-SOD) and total antioxidant capacity (T -AOC) levels. Western blotting was used to detect the expression of aquaporin 3 (AQP3) in cells. Results Compared with the normal group, the collagen density of the pollack skin collagen peptide was less than 300 mg · L -1 without cytotoxicity. The cell viability decreased to 54.0% (P <0.01) in H 2 O 2300μmol·L -1 group. Pre-treatment of 50, 200 and 200 μg · L -1 collagen peptide for 12 h resulted in 69.4%, 79.4% and 89.1% increase in cell viability (P <0.05, P <0.01). Compared with the model group, CAT, GSH-Px, T-SOD and T-AOC in 50, 200 and 200 mg · L -1 groups increased, The contents of CAT, GSH-Px, T-SOD and T-AOC increased 56.90%, 48.68%, 48.24% and 71.43% (P <0.01) correspondingly. Compared with the model group, the expression of AQP3 in the group of 200 mg · L -1 collagen decreased significantly to 67.1% (P <0.01). Conclusion Polypeptide of codfish skin collagen can protect H 2 O 2 -induced oxidative injury. The mechanism may be related to the enhancement of antioxidant capacity and the reduction of AQP3 expression.