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黄芪多糖对心肌过氧化氢损伤的保护机制研究

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摘要:

目的 探索黄芪多糖(APS)对过氧化氢(H2O2)诱发的大鼠心肌细胞H9C2的保护作用及可能机制。方法 实验分为①对照组;②对照+APS组;③H2O2损伤组;④H2O2损伤+APS修复组,采用乳酸脱氢酶(LDH)漏出量测定、3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]检测、免疫荧光染色、原位末端标记法(TUNEL)、逆转录-聚合酶链反应(RT-PCR)以及蛋白质印迹(WB)等实验方法,观察APS对H2O2引起的心肌细胞损伤、死亡的影响以及过氧化物酶体增殖物激活受体γ共激活因子1(PGC-1)/肌肉因子(irisin)信号通路在其中的参与情况。结果 与对照组相比,H2O2损伤组,细胞死亡率显著提高[(23.3±1.14)% vs.(12.81±0.71)%,P<0.001];与H2O2组相比,H2O2损伤+APS修复组黄芪多糖组能够显著降低过氧化氢引起的细胞死亡[(16.93±0.64)% vs. (23.3±1.14)%,P<0.001]、提高细胞生存率[(65.77±5.08)% vs. (39.95±4.90)%,P<0.05)]、减轻细胞凋亡[(18.90±1.14)% vs. (4.80±0.39)%,P<0.001]。黄芪多糖处理后,irisin及其上游调控因子PGC-1α的信使核糖核酸(mRNA)转录水平和蛋白表达量均明显提高,差异具有统计学意义(P<0.05)。结论 黄芪多糖对心肌细胞的保护作用至少部分是通过PGC-1α/irisin信号通路实现。

Abstract:

Objective This study explores the effect and mechanism of Astragalus polysaccharide in hydrogen peroxide-induced cardiomyocyte injury. Methods H9C2 cells were divided into four groups: ①control, ②control+APS group, ③H2O2 injury, ④H2O2+APS group. LDH leakage assay, MTT assay, immunofluorescence staining, TUNEL, RT-PCR, and western blot were employed to evaluate the effects of astragalus polysaccharides on cardiomyocyte apoptosis and death caused by hydrogen peroxide. PCR and western blot also examined PGC-1 α and irisin expression. Results Compared with the control group, the cell death rate after H2O2 admission increased from [(12.81±0.71)% vs. (23.3±1.14)%, P<0.001]. Astragalus polysaccharides could significantly reduce cell death caused by hydrogen peroxide [(16.93±0.64)% vs. (23.3±1.14)%, P<0.001], improve cell survival rate [(65.77 ±5.08)% vs. (39.95 ± 4.90)%, P<0.05], and inhibit cell apoptosis [(18.90±1.14)% vs. (4.80±0.39)%, P<0.001]. In addition, the mRNA and expression of irisin and its upstream regulator PGC-1α increased remarkably after astragalus polysaccharide treatment (P<0.05). Conclusion The protective effect of astragalus polysaccharides on cardiomyocytes is at least partly mediated by PGC-1 α/ Irisin signal path activation.

基金项目:

河南省医学科技攻关计划(联合共建项目) (LHGJ20191207)

参考文献:

  • 2008

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