hsa-miR-423-5p激活IL1R1基因在冠心病中的作用机制

Mechanism of hsa-miR-423-5p activating IL1R1 gene in coronary heart disease

  • 摘要: 目的:通过人群表达水平及转录组测序分析hsa-miR-423-5p靶向目标基因在冠心病(CHD)中的作用机制。方法:选取广西医科大学第一附属医院收治的151 例CHD 患者(CHD 组),同期选择健康体检者作为健康对照组。通过实时荧光定量PCR(RT-qPCR)检测外周血白细胞中hsa-miR-423-5p表达水平,通过Spearman相关分析对hsa-miR-423-5p与各临床指标的相关关系进行分析。在EA.hy 926细胞中过表达hsa-miR-423-5p后进行hsa-miR-423-5p的亚细胞定位实验及转录组测序。针对测序结果预测hsa-miR-423-5p的潜在靶基因,对其进行RT-qPCR实验验证并分析目标基因与hsa-miR-423-5p表达水平及各临床指标之间的相关关系。结果:CHD 组外周血白细胞中hsa-miR-423-5p 的表达水平显著低于健康对照组(P< 0.05)。hsamiR-423-5p 的表达量与总胆固醇(TC)呈负相关关系,与高密度脂蛋白胆固醇(HDL-C)呈正相关关系(P< 0.05)。在EA.hy 926细胞中过表达hsa-miR-423-5p,亚细胞定位实验显示hsa-miR-423-5p主要存在于细胞核内。转录组测序结果经过分析验证后,发现过表达hsa-miR-423-5p后可上调IL1R1表达。CHD组中IL1R1的表达水平低于健康对照组(P< 0.01),IL1R1表达水平与hsa-miR-423-5p表达水平、HDL-C均呈正相关关系(P< 0.01)。结论:hsa-miR-423-5p及IL1R1在CHD患者中呈低表达且与TC和HDL-C相关;hsa-miR-423-5p在内皮细胞上调IL1R1的表达进而影响血脂水平,提示hsa-miR-423-5p可能通过调控IL1R1基因影响脂质代谢及内皮细胞功能从而参与CHD的进展。

     

    Abstract: Objective:To analyze the mechanism of hsa-miR-423-5p target genes in coronary heart disease(CHD) by population expression level and transcriptome sequencing.Methods:A total of 151 CHD patients admitted to the First Affiliated Hospital of Guangxi Medical University were selected as the CHD group, and healthy physical examination subjects were selected as the healthy control group.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of hsa-miR-423-5p in peripheral blood leukocytes.Spearman correlation analysis was performed to analyze the correlation between hsa-miR-423-5p and various clinical indicators.Subcellular localization experiments and transcriptome sequencing were conducted after overexpression of hsa-miR-423-5p in EA.hy 926 cells.The sequencing results were used to predict the potential target genes of hsa-miR-423-5p, and RT-qPCR experiments were performed to validate and analyze the correlation between the target genes and hsa-miR-423-5p expression levels and various clinical indicators.Results:The expression level of hsa-miR-423-5p in peripheral blood leukocytes of the CHD group was significantly lower than that of the healthy control group(P< 0.05).Hsa-miR-423-5p expression level was negatively correlated with total cholesterol(TC)and positively correlated with high density lipoprotein cholesterol(HDLC) (P< 0.05).Subcellular localization experiments after overexpression of hsa-miR-423-5p in EA.hy 926 cells showed that hsa-miR-423-5p was mainly present in the nucleus.After transcriptome sequencing results were analyzed and validated, it was found that IL1R1 expression could be up-regulated after overexpression of hsa-miR-423-5p.The expression level of IL1R1 was lower in the CHD group than that in the healthy control group (P< 0.01), and IL1R1 expression level was positively correlated with the expression levels of hsa-miR-423-5p and HDL-C (P< 0.01).Conclusion:The expressions of hsa-miR-423-5p and IL1R1 are low in patients with CHD and correlated with TC and HDL-C.hsa-miR-423-5p up-regulates the expression of IL1R1 in endothelial cells, thereby affecting lipid levels, suggesting that hsa-miR-423-5p may affect lipid metabolism and endothelial cell function through the regulation of IL1R1 gene, thus participating in the development of CHD.

     

/

返回文章
返回