秦皮乙素调控miR-486-5p表达抑制高糖诱导的肾小管上皮细胞炎症损伤的机制研究

Mechanism of esculetin inhibiting high-glucose induced inflammatory injury of renal tubular epithelial cells by regulating expression of miR-486-5p

  • 摘要:
      目的  探讨秦皮乙素(Esc)对高糖诱导的肾小管上皮细胞炎症损伤的影响及分子机制。
      方法  肾小管上皮细胞HK-2随机分为对照(Con)组,高糖(HG)组,Esc低、中、高浓度(HG+Esc-L、M、H)组,HG+anti-miR-NC组,HG+anti-miR-486-5p组,HG+Esc+miR-NC组以及HG+Esc+miR-486-5p组。采用酶联免疫吸附法(ELISA)检测白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平;采用流式细胞术检测细胞凋亡率;采用实时荧光定量聚合酶链式反应(RT-qPCR)检测miR-486-5p表达水平。
      结果  Esc低、中、高浓度处理后,高糖诱导的HK-2细胞中IL-6、TNF-α水平及miR-486-5p表达水平逐渐降低,凋亡率逐渐降低,差异有统计学意义(P < 0.05)。抑制miR-486-5p表达后,高糖诱导的HK-2细胞中IL-6、TNF-α水平及细胞的凋亡率降低,差异有统计学意义(P < 0.05)。过表达miR-486-5p和Esc同时处理后,IL-6、TNF-α水平及细胞的凋亡率升高,差异有统计学意义(P < 0.05)。
      结论  Esc通过下调miR-486-5p表达抑制高糖诱导的肾小管上皮细胞炎症损伤。

     

    Abstract:
      Objective  To explore the effect of esculetin (Esc) on the inflammatory injury of renal tubular epithelial cells induced by high glucose and its molecular mechanism.
      Methods  Renal tubular epithelial cells HK-2 were randomly divided into control (Con) group, high glucose (HG) group, low, medium and high concentration of Esc (HG+Esc-L, M, H) groups, HG+anti-miR-NC group, HG+anti-miR-486-5p group, HG+Esc+miR-NC group and HG+Esc+miR-486-5p group. Enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels. Flow cytometry was used to detect cell apoptosis rate. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect miR-486-5p expression level.
      Results  After treatment with low, medium and high concentrations of Esc, the levels of IL-6 and TNF-α and miR-486-5p expression in HK-2 cells induced by high glucose were gradually decreased, while the apoptosis rate was gradually decreased, and there were significant differences between groups (P < 0.05). After inhibiting the expression of miR-486-5p, the levels of IL-6, TNF-α and the apoptosis rate in HK-2 cells induced by high glucose were decreased (P < 0.05). After simultaneous treatments with over-expression of miR-486-5p and Esc, IL-6 and TNF-α levels and cell apoptosis rate were increased (P < 0.05).
      Conclusion  Esc inhibits the inflammatory injury of renal tubular epithelial cells induced by high glucose by down-regulating the expression of miR-486-5p.

     

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