circMAT2B靶向miR-103a-3p对高糖诱导的肾小管上皮细胞损伤的影响

    Effect of circMAT2B on renal tubular epithelial cell damage induced by high glucose through targeting miR-103a-3p

      摘要
    • 摘要: 目的 探讨环状RNA MAT2B（circular RNA MAT2B，circMAT2B）靶向微小RNA-103a-3p（miR-103a-3p）对高糖诱导的肾小管上皮细胞损伤的影响及其作用机制。方法 体外培养人肾小管上皮细胞HK-2，采用高糖诱导HK-2细胞建立细胞损伤模型（HG组），正常培养的HK-2细胞记为Con组；si-NC、si-circMAT2B、miR-NC、miR-103a-3p mimics、si-circMAT2B与anti-miR-NC、si-circMAT2B与anti-miR-103a-3p分别转染入HK-2细胞后加入25 mmol/L葡萄糖处理细胞（HG+si-NC组、HG+si-circMAT2B组、HG+miR-NC组、HG+miR-103a-3p组、HG+si-circMAT2B+anti-miR-NC组、HG+si-circMAT2B+anti-miR-103a-3p组）；采用定量反转录聚合酶链式反应法检测circMAT2B与miR-103a-3p的表达量；采用酶联免疫吸附法检测细胞培养上清液白细胞介素6（interleukin-6，IL-6）与肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）的水平；采用流式细胞术检测细胞凋亡率；双荧光素酶报告基因实验检测circMAT2B与miR-103a-3p的靶向关系。结果 与Con组比较，HG组circMAT2B的表达水平与凋亡率升高（P<0.05），IL-6、TNF-α的水平升高（P<0.05），miR-103a-3p的表达水平降低（P<0.05）；与HG+si-NC组比较，HG+si-circMAT2B组IL-6、TNF-α的水平降低（P<0.05），凋亡率降低（P<0.05）；circMAT2B能够吸附miR-103a-3p而靶向调控其表达；与HG+miR-NC组比较，HG+miR-103a-3p组IL-6、TNF-α的水平与Bax蛋白水平降低（P<0.05），凋亡率降低（P<0.05）；与HG+si-circMAT2B+anti-miR-NC组比较，HG+si-circMAT2B+anti-miR-103a-3p组IL-6、TNF-α的水平升高（P<0.05），凋亡率升高（P<0.05）。结论 抑制circMAT2B表达可通过负向调控miR-103a-3p的表达而抑制细胞炎症因子的分泌及凋亡从而减轻高糖诱导的肾小管上皮细胞损伤。

       

      Abstract: Objective To explore the effect of circMAT2B targeting miR-103a-3p on renal tubular epithelial cell damage induced by high glucose and elucidate its mechanism.Methods Human renal tubular epithelial cells HK-2 were cultured in vitro and HK-2 cells induced by high glucose for modeling cell injury (HG group).Normally cultured HK-2 cells were recorded as Con group.And si-NC,si-circMAT2B,miR-NC,miR-103a-3p mimics,si-circMAT2B,anti-miR-NC,si-circMAT2B and anti-miR-103a-3p were transfected into HK-2 cells and then added 25 mmol/L glucose-treated cells (HG+si-NC group,HG+si-circMAT2B group,HG+miR-NC group,HG+miR-103a-3p group,HG+si-circMAT2B+anti-miR-NC group,HG+si-circMAT2B+anti-miR-103a-3p group).Quantitative real-time polymerase chain reaction (qRT-PCR)was utilized for detecting the expression of circMAT2B and miR-103a-3p;Enzyme-linked immunosorbent assay (ELISA0 for detecting the levels of IL-6 and TNF-α;Flow cytometry for detecting apoptotic rate;Dual luciferase reporter gene experiment for examining the targeting relationship between circMAT2B and miR-103a-3p.Results As compared with Con group,the expression level of circMAT2B and apoptotic rate rose in HG group (P<0.05),the levels of IL-6 and TNF-α spiked (P<0.05)while the expression level of miR-103a-3p dropped (P<0.05).As compared with HG+si-NC group,the levels of IL-6 and TNF-α declined in HG+si-circMAT2B group (P<0.05) and apoptotic rate dropped (P<0.05).circMAT2B could adsorb miR-103a-3p and target its expression.Compared with HG+miR-NC group,the levels of IL-6 and TNF-α declined in HG+miR-103a-3p group (P<0.05)while apoptotic rate dipped (P<0.05).As compared with HG+si-circMAT2B+anti-miR-NC group,the levels of IL-6,TNF-α rose in HG+si-circMAT2B+anti-miR-103a-3p group (P<0.05) and apoptotic rate rose (P<0.05).Conclusions An inhibition of circMAT2B expression may inhibit the secretion and apoptosis of cytoinflammatory factors by negatively regulating the expression of miR-103a-3p,thereby reducing the damage of renal tubular epithelial cells induced by high glucose.

       

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