刘静, 李晓莉, 陈蕊蕊, 魏婷, 艾永飞, 李炜, 刘慧. IRE1α调节miRNA34a在高糖诱导的心肌细胞肥大中的作用[J]. 心脏杂志, 2019, 31(4): 373-378. DOI: 10.12125/j.chj.201811034
    引用本文: 刘静, 李晓莉, 陈蕊蕊, 魏婷, 艾永飞, 李炜, 刘慧. IRE1α调节miRNA34a在高糖诱导的心肌细胞肥大中的作用[J]. 心脏杂志, 2019, 31(4): 373-378. DOI: 10.12125/j.chj.201811034
    Jing LIU, Xiao-li LI, Rui-rui CHEN, Ting WEI, Yong-fei AI, Wei LI, Hui LIU. Role of IRE1α in regulating miRNA34a in high glucose-induced cardiomyocyte hypertrophy[J]. Chinese Heart Journal, 2019, 31(4): 373-378. DOI: 10.12125/j.chj.201811034
    Citation: Jing LIU, Xiao-li LI, Rui-rui CHEN, Ting WEI, Yong-fei AI, Wei LI, Hui LIU. Role of IRE1α in regulating miRNA34a in high glucose-induced cardiomyocyte hypertrophy[J]. Chinese Heart Journal, 2019, 31(4): 373-378. DOI: 10.12125/j.chj.201811034

    IRE1α调节miRNA34a在高糖诱导的心肌细胞肥大中的作用

    Role of IRE1α in regulating miRNA34a in high glucose-induced cardiomyocyte hypertrophy

    • 摘要:
        目的  研究肌醇需求因子(inositol-requiring enzyme,IRE)1α在高糖诱导的心肌细胞肥大中的作用及与miRNA34a的关系。
        方法  分离培养心肌细胞并分为对照组,高糖组,对照+过表达IRE1α组,高糖+过表达IRE1α组。利用IRE1α的腺病毒干预细胞,观察高糖条件下其对心肌细胞活力、肥大及心钠肽(ANP)表达的影响。Western blot检测ANP和IRE1α的表达,qRT-PCR检测ANP、IRE1α和miRNA34a表达,免疫荧光染色检测细胞纯度及横截面积。
        结果  与对照组比较,随着葡萄糖浓度上升,细胞活力持续下降(P<0.05),心肌细胞肥大标志ANP表达增加(P<0.05),IRE1α基因及蛋白表达降低(P<0.05),且于葡萄糖浓度达到30 mmol/L最低。与对照组比较,高糖能够降低细胞活力,刺激心肌细胞上调ANP表达(P<0.05),刺激心肌细胞横截面积增大(P<0.05);而过表达IRE1α基因能显著上调心肌细胞的活力水平,降低ANP蛋白及基因的表达,抑制高糖培养下心肌细胞的肥大(P<0.05)。qRT-PCR结果显示,随着血糖浓度上升,miRNA34a表达明显升高,且当过表达IRE1α后,miRNA34a表达明显降低。
        结论  过表达IRE1α基因可有效抑制高糖诱导的心肌细胞肥大,其机制可能与下调miRNA34a表达有关。

       

      Abstract:
        AIM  To study the role of inositol-requiring enzyme1 α (IRE1 α) gene in cardiomyocyte hypertrophy induced by high glucose and its relationship with miRNA34a.
        METHODS  Cardiomyocytes were isolated and cultured. They were divided into different groups: a control group, a high glucose overexpression IRE1α group, and s control IRE1α group (Con Ad-IRE1α). IRE1α adenovirus was used to interfere with these cells to observe the effects of high glucose on cardiomyocyte viability, hypertrophy, and the expression of atrial natriuretic peptide (ANP). Western blot was used to detect the expression of ANP and IRE1α. qRT-PCR was utilized to detect the expression of ANP, IRE1α, and miRNA34a. Immunofluorescence staining was utilized to detect cell purity and cross-sectional area.
        RESULTS  Compared with the control group, with the increase of glucose concentration, the cell viability continued to decrease, the expression of ANP increased, the expression of IRE1α gene and protein decreased, and the glucose concentration reached a lowest level of 30 mmol/L. Compared with the control group, high glucose decreased the cell viability, stimulated the expression of ANP and increased the cross sectional area of the cardiomyocytes (P < 0.05), while over-expression of IRE1α gene significantly up-regulated the levels of cell viability of cardiomyocytes. The expression of ANP protein and gene decreased and hypertrophy of cardiomyocytes was inhibited in the high glucose culture (P < 0.05). In addition, qRT-PCR results showed that miRNA34a expression gradient increased with the increase of high glucose concentration and miRNA34a expression decreased significantly after over-expression of IRE1α gene.
        CONCLUSION  Over-expression of IRE1α gene effectively inhibits hypertrophy induced by high glucose and its mechanism may be related to down-regulation of miRNA34a expression.

       

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