西番莲果皮花色苷对H2O2诱导L02细胞氧化损伤的抑制作用

Inhibition effect of peel of passion fruits anthocyanins on H2O2-induced oxidative damage in L02 cells

中文关键词:  西番莲果皮  花色苷  L02细胞  抗氧化活性  非靶向代谢组学

英文关键词:peel of passion fruits  anthocyanins  L02 cell  antioxidant activity  non-targeted metabolomics

中文摘要:

      目的 探究西番莲果皮花色苷(peel of passion fruits anthocyanins, PPFA)对H2O2诱导人正常肝细胞L02氧化损伤的抑制作用。方法 采用噻唑蓝[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT]法建立H2O2诱导L02细胞氧化损伤的模型, 通过测定细胞丙二醛(malondialdehyde, MDA)、乳酸脱氢酶(lactate dehydrogenase, LDH)、超氧化物歧化酶(superoxide dismutase, SOD)、过氧化氢酶(catalase, CAT)和谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)指标, 观察PPFA对细胞氧化损伤的抑制作用, 结合非靶向代谢组学显著差异代谢物分析, 探讨其抗氧化机制。结果 PPFA体外清除1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl, DPPH)自由基的半抑制浓度(half maximal inhibitory concentration, IC50)值为34.62 μg/mL, 有一定的羟自由基、超氧阴离子自由基清除能力。细胞实验表明, PPFA能够显著提高H2O2损伤的L02细胞总抗氧化力(P<0.05), 并显著降低损伤细胞中MDA生成量和LDH的释放量(P<0.05); 与损伤组相比, PPFA各剂量组SOD活性均显著升高(P<0.05), 中剂量组CAT与低剂量组GSH-Px的活性显著升高且达空白组水平(P<0.05)。代谢组学分析表明, 与损伤组相比, PPFA处理后细胞存在23种显著差异代谢物, 涉及到谷胱甘肽代谢、胆汁分泌、铁死亡和脂肪酸生物合成途径。结论 PPFA对H2O2诱导L02细胞的氧化损伤有抑制作用, 表现出显著的细胞抗氧化活性。

英文摘要:

      Objective To investigate the inhibitory effect of peel of passion fruits anthocyanins (PPFA) on H2O2-induced oxidative damage in human normal liver cells L02. Methods A model of H2O2-induced oxidative damage in L02 cells was established by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method, and the inhibitory effect of PPFA on cellular oxidative damage was observed by measuring cellular malondialdehyde (MDA), lactate dehydrogenase (LDH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) indexes, combined with non-targeted metabolomics significantly different metabolite analysis to explore its antioxidant mechanism. Results The half maximal inhibitory concentration (IC50) value of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging by PPFA in vitro was 34.62 μg/mL, and it had certain scavenging ability hydroxyl radicals and superoxide anion radicals. Cellular experiments showed that PPFA could significantly increase the total antioxidant power of H2O2-induced L02 cells (P<0.05), and significantly decrease the MDA production and LDH release in the damaged cells (P<0.05); compared with the damaged group, the SOD activity of PPFA was significantly increased in all dose groups (P<0.05), and the activity of CAT in the middle dose group and GSH-Px in the low dose group were significantly increased and reached the normal level (P<0.05). Metabolomic analysis showed that there were 23 kinds of significantly different metabolites in PPFA-treated cells compared with the damaged group, involving glutathione metabolism, bile secretion, ferroptosis and fatty acid biosynthesis pathways. Conclusion PPFA can inhibit the H2O2-induced oxidative damage in L02 cells, showing significant cellular antioxidant activity.