ZHENG Feng,JIN Fang-duo,JIN Mei-hua,et al.Protective Effect of Soy Isoflavones on Injury of L02 Cells Induced by H2O2[J].Soybean Science,2020,39(03):458-463.[doi:10.11861/j.issn.1000-9841.2020.03.0458]
大豆异黄酮对H2O2致L02细胞损伤的保护作用
- Title:
- Protective Effect of Soy Isoflavones on Injury of L02 Cells Induced by H2O2
- Keywords:
- Soybean; Isoflavones; L02; H2O2; Protective effect
- 文献标志码:
- A
- 摘要:
- 为探究大豆异黄酮（soy isoflavones，ISOF）对过氧化氢（H2O2）诱导的L02细胞损伤的保护作用，使用ISOF对L02对数生长期细胞进行预保护，再用H2O2诱导L02细胞氧化损伤，建立细胞损伤模型。采用CCK-8法检测L02细胞存活率，采用微板法测定L02细胞培养液中谷丙转氨酶（ALT）、谷草转氨酶（AST）、乳酸脱氢酶（LDH）活性以及细胞内还原型谷胱甘肽（GSH）含量，采用羟胺法测定细胞中超氧化物歧化酶（SOD）活性，采用硫代巴比妥酸（TBA）法检测细胞丙二醛（MDA）水平。结果显示：300 μmol?L-1 H2O2处理L02细胞2 h能够使L02细胞存活率降低51%，并显著增高L02细胞LDH向细胞培养液的漏出，说明H2O2处理已造成L02细胞的损伤。ISOF在质量浓度10~40 mg?L-1时，对L02细胞无细胞毒作用，是安全的。安全剂量ISOF预处理能够改变L02细胞的上述损伤指标，与H2O2损伤组比较，40 mg?L-1 ISOF组L02细胞存活率增高28.9%（P<0.05），LDH、ALT和AST漏出率分别降低91.4%、91.7%和74.1%，细胞MDA生成量降低118.5%，GSH含量和SOD活性分别增高184.4%和76.2%。结果表明ISOF能保护H2O2诱导的L02细胞氧化损伤。
- Abstract:
- In order to detect the protective effect of soy isoflavones (ISOF) on injury of L02 cells induced by hydrogen peroxide (H2O2), this study established the cellular injury model after the L02 cells were pretreatment with ISOF and induced with H2O2. The cell viability was investigated with cell counting kit-8 (CCK-8) assay, the medium activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and the cellular reduced glutathione (GSH) were tested with the microplate method, superoxide dismutase (SOD) activity was assayed with hydroxylamine method, the malondialdelyde (MDA) content was tested with the thibabituric acid (TBA) method. The results showed that H2O2 decreased the viability of L02 cells by 51%, and elevated the LDH leakage to culture medium, indicative of the establishment of oxidative injury model of L02 cells. ISOF had no cytotoxicity within the scope of the concentration of 10-40 mg?L-1. However, pretreatment with ISOF could alleviate the above-mentioned damage indicators of L02 cells. Compared with H2O2 injury group, the cell viability of L02 was increased by 28.9% (P<0.05), the leakage of LDH, ALT, AST was reduced by 91.7%, 91.7% and 74.1%, respectively, the cellular MDA was decreased by 118.5%, and the cellular GSH and SOD were increased by 184.4% and 76.2% respectively in the 40 mg?L-1 ISOF group. It is suggested that ISOF could protect L02 cells from oxidative damage induced by H2O2.
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