丝素肽对过氧化氢致A549细胞损伤的修复作用
Restoration Effect of Silk Fibroin Peptide on Hydrogen Dioxide Induced Human Lung Cancer Cell Injury
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摘要:
目的 探究丝素肽（SF）对过氧化氢（H2O2）致人肺癌（A549）细胞损伤的修复作用。
方法 将细胞分为空白对照组，H2O2组（600 μmol/L），SF前处理组（10、20、30、50 mg/mL SF预孵育24 h再加600 μmol/L H2O2损伤24 h），SF后处理组（600 μmol/L H2O2损伤24 h再加10、20、30、50 mg/mL SF孵育24 h）及阳性对照前、后处理组抗氧化剂N-乙酰半胱氨酸（NAC）。用MTT法测定细胞活力，生物化学法测定细胞中丙二醛（MDA）、超氧化物岐化酶（SOD）、过氧化氢酶（CAT）的含量和总抗氧化能力（T-AOC）。
结果 H2O2组细胞活力为（46.67±2.19）%，MDA含量为（64.31±3.22）nmol/mg（以蛋白计）。与H2O2组相比，前处理组和后处理组中细胞活力增加，而MDA含量降低（
P < 0.05）；SF后处理组细胞活力的增加和MDA含量的降低更明显（P < 0.05）；尤其50 mg/mL SF后处理组比前处理组细胞活力增加值高2.33%，而MDA含量降低值少44.51 nmol/mg（以蛋白计）。H2O2组中SOD、CAT活性和T-AOC都明显降低（P < 0.05），SF后处理明显提高SOD、CAT活性和T-AOC水平（P < 0.05）。结论 SF对H2O2致A549细胞损伤有修复作用，部分原因可能是由于其可增加细胞活力和提高细胞中抗氧化酶活性。
Abstract:Objective To assess the restoration effect of silk fibroin peptide (SF) on hydrogen dioxide (H2O2) induced human lung cancer cell (A549) injury.
Methods The A549 cells were divided into blank (without drug treatment), H2O2 (600 μmol/L), pretreatment (10, 20, 30, and 50 mg/mL SF pretreatment for 24 h plus 600 μmol/L H2O2 treatment for another 24 h), post-treatment (600μmol/L H2O2 treatment for 24 h and 10, 20, 30, and 50 mg/mL SF treatment for another 24 h), and positive control pretreatment and post-treatment groups antioxidant, N-acetylcysteine (NAC). Cell viability was measured by MTT assay, and levels of malondialdehyde (MDA), super oxide dismutase (SOD), catalase (CAT), and total antioxidant capacity (T-AOC) by biochemical methods.
Results The cell viability was (46.67±2.19)% and the content of MDA was (64.31±3.22)nmol/mg (protein) after H2O2 treatment. Compared with the H2O2 group, the cell viability increased and the content of MDA decreased (
P < 0.05) in the pretreatment groups and the post-treatment groups, and the post-treatment groups showed greater effects (P < 0.05). Especially when comparing the 50 mg/mL SF treatment effects, post-treatment increased cell viability by 2.33% and decreased MDA content by 44.51 nmol/mg (protein) than the pretreatment did. The activities of SOD, CAT, and T-AOC were all decreased in the H2O2 group (P < 0.05), but the activities were increased after post-treatment with SF (P < 0.05).Conclusion The study demonstrates that SF could promote cell viability and intracellular antioxidase activity to ameliorate A549 cell injury induced by H2O2.
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