甘肃中医药大学学报2025，Vol.42Issue(6)：17-23,7.DOI:10.16841/j.issn1003-8450.2025.06.03
通脉消斑汤对大鼠心肌细胞缺氧复氧后N6-甲基腺苷RNA修饰及炎症通路的调节作用
Regulatory effect of Tongmai Xiaoban Tang(通脉消斑汤)on N6-methyladenosine RNA modification and inflammatory pathway in rat cardiomyocytes following hypoxia-reoxygenation
摘要
Abstract
Objective To investigate the protective effect of Tongmai Xiaoban Tang(通脉消斑汤,TMXBT)on hypoxia-reoxygenation injury in rat cardiomyocytes and its regulatory mechanisms on N6-methyladenosine(m6A)RNA modification and inflammatory pathways.Methods Rat cardiomyocyte line H9C2 was divided into four groups:control group,hypoxia group,hypoxia-reoxygenation group,and Tongmai Xiaoban Tang group.A hypoxia-reoxygenation injury model was established,with the Tongmai Xiaoban Tang group receiving Tongmai Xiaoban Tang intervention during reoxygenation.Cell proliferation and cytotoxicity were assessed using Cell Counting Kit-8(CCK-8).m6A RNA methylation levels were quantified via an m6A RNA methylation assay kit.Nuclear translocation of phosphorylated nuclear factor kappa B p65 subunit(p-NF-κB p65)was observed by immunofluorescence staining.Protein expression of fat mass and obesity-associated protein(FTO),p-NF-κB p65,tumor necrosis factor-alpha(TNF-α),and interleukin-6(IL-6)was measured using Western Blot.Results Compared with control group,hy-poxia group exhibited significantly increased cell inhibition rate,m6A RNA levels,and p-NF-κB p65,IL-6 protein expression levels(P＜0.05 or P＜0.01),while FTO protein expression level decreased(P＜0.05).Hypoxia-reoxy-genation group significantly elevated cell inhibition rate,m6A RNA levels,and FTO,p-NF-κB p65,TNF-α pro-tein expression levels compared to hypoxia group(P＜0.05).Tongmai Xiaoban Tang intervention significantly re-duced cell inhibition rate,m6A RNA levels,and p-NF-κB p65,TNF-α,IL-6 protein expression levels compared to hypoxia-reoxygenation group(P＜0.05 or P＜0.01),alongside increased FTO protein expression level(P＜0.05).Immunofluorescence staining revealed that weak nuclear expression of p-NF-κB p65 protein was observed in the control group.In contrast,nuclear expression of this protein was significantly elevated in both the hypoxia group and the hypoxia-reoxygenation group.Notably,the nuclear expression level of p-NF-κB p65 protein in the Tongmai Xiaoban Tang group was significantly lower than that in the hypoxia-reoxygenation group.Conclusion Tongmai Xiaoban Tang mitigates hypoxia-reoxygenation injury in rat cardiomyocytes by upregulating FTO protein expression to reduce m6A RNA methylation levels,suppressing NF-κB inflammatory pathway activation,and inhibiting down-stream inflammatory cytokine release,thereby exerting cardioprotective effects.关键词
心肌损伤/缺氧复氧/通脉消斑汤/N6-甲基腺苷/炎症通路/实验研究Key words
myocardial injury/hypoxia-reoxygenation/Tongmai Xiaoban Tang(通脉消斑汤)/N6-methylade-nosine(m6A)/inflammatory pathway/experimental study分类
医药卫生引用本文复制引用
赵海梅,程红,焦萁荟..通脉消斑汤对大鼠心肌细胞缺氧复氧后N6-甲基腺苷RNA修饰及炎症通路的调节作用[J].甘肃中医药大学学报,2025,42(6):17-23,7.基金项目
深圳市科技计划项目(JCYJ20220531091817039). （JCYJ20220531091817039）
