本文選題：微小RNA　切入點(diǎn)：糖尿病性視網(wǎng)膜病變　出處：《華中科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年04期 　論文類型：期刊論文

【摘要】：目的探討miR-15a對糖尿病性視網(wǎng)膜病變抗炎和抗新生血管生成的調(diào)節(jié)作用。方法通過25mmol/L葡萄糖誘導(dǎo)人視網(wǎng)膜色素上皮細(xì)胞株HARPE-19建立高糖模型(HG),并以5mmol/L葡萄糖誘導(dǎo)的HARPE-19細(xì)胞作為對照(LG),將HG組和LG組分為4個亞組:miR-15amimic組、NC-mimic組、miR-15ainhibitor組和NC-inhibitor組,MTT增殖實(shí)驗(yàn)檢測細(xì)胞增殖能力,細(xì)胞劃痕實(shí)驗(yàn)檢測細(xì)胞遷移能力,雙熒光素酶報(bào)告基因?qū)嶒?yàn)驗(yàn)證miR-15a與酸性鞘磷脂酶(ASM)的關(guān)系,RT-PCR檢測miR-15a、ASM、VEGF、IL-1β、IL-6、TNF-αmRNA表達(dá)。結(jié)果 NC-mimic細(xì)胞和NC-inhibitor細(xì)胞中,HG組miR-15a表達(dá)均顯著低于LG組(均P0.05)。miR-15amimic組ASM mRNA表達(dá)水平顯著低于NC-mimic組(P0.05),miR-15ainhibitor組ASM mRNA表達(dá)水平顯著高于NC-inhibitor組(P0.05)。miR-15amimic組細(xì)胞遷移距離顯著大于NC-mimic組(P0.05),miR-15ainhibitor組細(xì)胞遷移距離顯著小于NC-inhibitor組(P0.05)。轉(zhuǎn)染24~120h,MTT法檢測miR-15amimic組吸光度值顯著高于NC-mimic組(P0.05),miR-15ainhibitor組吸光度值顯著低于NC-inhibitor組(P0.05)。NC-mimic細(xì)胞和NC-inhibitor細(xì)胞中,HG組VEGF、IL-1β、IL-6、TNF-αmRNA表達(dá)水平均顯著高于LG組(均P0.05)。結(jié)論降低miR-15a表達(dá)能夠促進(jìn)視網(wǎng)膜細(xì)胞炎癥反應(yīng)和血管新生,miR-15a通過炎癥反應(yīng)和血管生成雙重調(diào)節(jié)作用參與糖尿病性視網(wǎng)膜病變的發(fā)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- 偽 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.
【作者單位】： 四川省西南醫(yī)科大學(xué)附屬醫(yī)院眼科;四川省德陽市人民醫(yī)院眼科;
【基金】：四川省科學(xué)技術(shù)廳科研基金資助項(xiàng)目(No.15ZC0869)
【分類號】：R587.2;R774.1

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