红花黄色素对心肌细胞H9C2的保护研究

Protective effect of safflor yellow on cardiomyocyte H9C2

目的探讨红花黄色素调控缺氧/复氧后心肌细胞对H9C2氧化应激的潜在机制。方法红花黄色素处理缺氧/复氧后心肌细胞H9C2后,检测其氧化应激指标丙二醛(MDA),过氧化氢酶(CAT),超氧化物歧化酶(SOD),谷胱甘肽过氧化物酶(GSH-Px)的水平。实时荧光定量PCR检测氧化应激关键基因的表达水平。RNA-seq检测红花黄色素处理后心肌细胞H9C2的miRNA表达差异。表达差异miRNA检测氧化应激关键基因的表达水平。结果红花黄色素处理缺氧/复氧后心肌细胞H9C2后,MDA的水平下降(P<0.05),SOD、CAT、GSH-Px的水平上升(P<0.05)。红花黄色素处理缺氧/复氧后心肌细胞H9C2后,氧化应激关键基因MPO的表达水平下降(P<0.05),SOD2,CAT,谷胱甘肽过氧化物酶4(GPX4)的表达水平上升(P<0.05)。过表达hsa-miR-887-5p时MPO的表达水平下降(P<0.05)、过表达hsa-miR-382-5p时SOD2的表达水平下降(P<0.05)、过表达hsa-miR-639时CAT2的表达水平下降(P<0.05)、过表达hsa-miR-761时GPX4的表达水平下降(P<0.05)。结论红花黄色素通过调控hsa-miR-382-5p、hsa-miR-887-5p、hsa-miR-639、hsa-miR-761的表达水平,分别靶向SOD2,MPO,CAT,GPX4 mRNA的3端非编码区,之后调控其表达水平,最终调节缺氧/复氧后心肌细胞H9C2的氧化应激。

Objective To discuss the potential mechanism of safflor yellow regulation in oxidative stress of cardiomyocyte H9C2 after hypoxia/reoxygenation(H/R).Methods After safflor yellow treating H/R cardiomyocyte H9C2,the levels of malondialdehyde(MDA),catalase(CAT),superoxide dismutase(SOD)and glutathione peroxidease(GSH-Px)were detected.The expressions of key genes of oxidative stress were detected by using RT-qPCR.The expression difference of cardiomyocyte H9C2 miRNA treated by safflor yellow was detected by using RNA-seq.The expressions of key genes of oxidative stress were detected by using miRNA with differential expression.Results After H/R cardiomyocyte H9C2 treated by safflor yellow,MDA decreased(P<0.05)and SOD,CAT and GSH-Px increased(P<0.05),and the expression of myeloperoxidase(MPO,key gene of oxidative stress)decreased(P<0.05)and superoxide dismutase 2(SOD2),CAT and glutathione peroxidase 4(GPX4)increased(P<0.05).The expression of MPO decreased in over-expressed hsa-miR-887-5p(P<0.05),and SOD2 expression decreased in over-expressed hsa-miR-382-5p(P<0.05).The expression of CAT2 decreased in over-expressed hsa-miR-639(P<0.05),and GPX4 expression decreased in over-expressed hsa-miR-761(P<0.05).Conclusion Safflower yellow can target the 3-terminal non-coding regions of SOD2,MPO,CAT,and GPX4 mRNAs respectively through regulating the expressions of hsa-miR-382-5p,hsa-miR-887-5p,hsa-miR-639,and hsa-miR-761,and then regulate their expression levels and finally regulate the oxidative stress of H/R cardiomyocyte H9C2.