miR-15a对糖尿病性视网膜病变抗炎和抗新生血管生成的双重调节作用

发布时间：2018-03-14 21:38

本文选题：微小RNA　切入点：糖尿病性视网膜病变　出处：《华中科技大学学报(医学版)》2017年04期 　论文类型：期刊论文

【摘要】：目的探讨miR-15a对糖尿病性视网膜病变抗炎和抗新生血管生成的调节作用。方法通过25mmol/L葡萄糖诱导人视网膜色素上皮细胞株HARPE-19建立高糖模型(HG),并以5mmol/L葡萄糖诱导的HARPE-19细胞作为对照(LG),将HG组和LG组分为4个亚组:miR-15amimic组、NC-mimic组、miR-15ainhibitor组和NC-inhibitor组,MTT增殖实验检测细胞增殖能力,细胞划痕实验检测细胞迁移能力,双荧光素酶报告基因实验验证miR-15a与酸性鞘磷脂酶(ASM)的关系,RT-PCR检测miR-15a、ASM、VEGF、IL-1β、IL-6、TNF-αmRNA表达。结果 NC-mimic细胞和NC-inhibitor细胞中,HG组miR-15a表达均显著低于LG组(均P0.05)。miR-15amimic组ASM mRNA表达水平显著低于NC-mimic组(P0.05),miR-15ainhibitor组ASM mRNA表达水平显著高于NC-inhibitor组(P0.05)。miR-15amimic组细胞迁移距离显著大于NC-mimic组(P0.05),miR-15ainhibitor组细胞迁移距离显著小于NC-inhibitor组(P0.05)。转染24~120h,MTT法检测miR-15amimic组吸光度值显著高于NC-mimic组(P0.05),miR-15ainhibitor组吸光度值显著低于NC-inhibitor组(P0.05)。NC-mimic细胞和NC-inhibitor细胞中,HG组VEGF、IL-1β、IL-6、TNF-αmRNA表达水平均显著高于LG组(均P0.05)。结论降低miR-15a表达能够促进视网膜细胞炎症反应和血管新生,miR-15a通过炎症反应和血管生成双重调节作用参与糖尿病性视网膜病变的发生。
[Abstract]:Objective to investigate the effects of miR-15a on anti-inflammatory and anti-angiogenesis of diabetic retinopathy. Methods High glucose glucose induced human retinal pigment epithelium cell line HARPE-19 was used to establish a high-glucose model, and HARPE-19 was induced by 5 mmol / L glucose. The HG group and LG group were divided into 4 subgroups: 1 miR-15amimic group, NC-mimiic group and NC-inhibitor group. The proliferative ability of HG group and LG group was detected by MTT assay. The cell migration ability was measured by cell scratch assay. Double luciferase reporter gene experiment confirmed the relationship between miR-15a and acid sphingolipase (ASM). RT-PCR was used to detect the expression of ASM mRNA in NC-mimic cells and NC-inhibitor cells. Results the expression of miR-15a in NC-mimic cells and NC-inhibitor cells was significantly lower than that in LG group (P 0.05. MiR-15amimic group) and ASM mRNA expression level was significantly lower than that in NC-mimic group (P0.05. MiR-15amimic group). The expression of ASM mRNA in group P0.05 miR-15ainhibitor was significantly higher than that in group NC-inhibitor (P0.05) .miR-15amimic group was significantly longer than that in group NC-mimic (P 0.05%). The absorbance of miR-15amimic group was significantly higher than that of NC-mimic group (P 0.05 miR-15aininhibitor group), and the absorbance of miR-15amimic group was significantly higher than that of NC-mimic group (P 0.05 miR-15aininhibitor group). Compared with NC-inhibitor group, the level of VEGFU IL-1 尾 IL-6 TNF- 伪 mRNA in HG group was significantly higher than that in LG group (P 0.05). Conclusion the decrease of miR-15a expression can promote the inflammatory response of retinal cells and the regulation of angiogenesis through inflammation and angiogenesis. It is involved in the development of diabetic retinopathy.
【作者单位】：四川省西南医科大学附属医院眼科;四川省德阳市人民医院眼科;
【基金】：四川省科学技术厅科研基金资助项目(No.15ZC0869)
【分类号】：R587.2;R774.1

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