葶苈子中抗心肌细胞凋亡成分筛选及机制研究

Screening of anti-cardiomyocyte apoptosis components in Tinglizi and study of mechanism

关键词:  葶苈子  北葶新苷  心肌损伤  细胞凋亡  氧化应激

Key words:  Tinglizi  lepidiumoside  cardiomyocyte injury  apoptosis pathway  oxidative stress

摘要:

目的? 从葶苈子中提取分离的大量化合物中筛选对H2O2诱导的心肌细胞损伤具有改善作用的成分,初步探讨其作用机制。方法? 用200 μmol·L-1 H2O2建立H9c2心肌细胞损伤模型,用葶苈子中化合物处理细胞6 h,检测细胞活力,氧化应激,凋亡水平和凋亡通路关键蛋白。结果? 葶苈子中10种化合物可以有效提升心肌细胞活力。山奈酚-3-O-β-D-吡喃葡萄糖苷(DS-1)、槲皮素-3-O-β-D-吡喃木糖苷(DS-2)和北葶新苷A(LS-19)降低心肌细胞凋亡率,升高总超氧化物歧化酶(total superoxide dismutase, T-SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-PX)水平,降低乳酸脱羟酶(lactate dehydrogenase, LDH)、丙二醛(malondialdehyde, MDA)水平,改善氧化应激；提高线粒体膜电位水平,降低活性氧(reactive oxygen species, ROS)水平,抑制细胞凋亡；降低 Bcl-2相关X蛋白(Bcl-2-associated X protein, Bax)/B淋巴细胞瘤-2(B lymphocytoma-2, Bcl-2),抑制凋亡通路关键蛋白半胱氨酸天冬氨酸蛋白酶-3(Cysteine aspartate protease-3, Caspase-3)表达水平。结论 葶苈子中化合物DS-1、DS-2和LS-19对H2O2诱导的H9c2细胞损伤具有显著保护作用, 机制可能与改善氧化应激稳态,抑制细胞凋亡有关。

Abstract:

OBJECTIVE To screen components with ameliorative effects on H2O2-induced cardiomyocyte injury from a large number of compounds extracted and isolated from Tinglizi and initially to investigate their mechanisms of action. METHODS The H9c2 cardiomyocyte injury model was induced by 200 μmol·L-1 H2O2, and the cells were treated with monomer compounds from Tinglizi for 6 h, then measured cell viability, oxidative stress, apoptosis levels and key proteins of the apoptosis pathway. RESULTS Ten compounds in Tinglizi could effectively enhance cell viability. Further research showed Kaempferol-3-O-β-D-glucopyranoside (DS-1), quercetin-3-O-β-D-xylopyranoside (DS-2) and lepidiumoside A (LS-19) can reduce the apoptosis rate of cardiomyocyte, enhance total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-PX) levels, decrease lactate dehydrogenase (LDH), malondialdehyde (MDA) levels, improve oxidative stress, enhance mitochondrial membrane potential level, decrease reactive oxygen species (ROS) level, and inhibit apoptosis of cardiomyocytes, reduce the level of Bcl-2-associated X protein (Bax)/B lymphocytoma-2 (Bcl-2), and inhibit the expression level of the key protein Cysteine aspartate protease-3 (Caspase-3) of the apoptosis pathway. CONCLUSION DS-1, DS-2 and LS-19 from Tinglizi have significant protective effect on H9c2 cardiomyocyte in H2O2-induced injury, the mechanism may be related to improve oxidative stress homeostasis the and inhibit the apoptotic pathway.