抑制长链非编码RNA-C920021L13Rik靶向miR-2861减轻糖尿病肾脏疾病系膜细胞损伤

孙琳琳; 朱鼎玉; 陈富华; 丁淼; 谢心苗

doi:10.3969/j.issn.1671-2390.2024.02.007

抑制长链非编码RNA-C920021L13Rik靶向miR-2861减轻糖尿病肾脏疾病系膜细胞损伤

lncRNA-C920021L13Rik depletion lessened high glucose-induced mesangial cell injury through regulating miR-2861

摘要

摘要:
目的探讨长链非编码RNA C920021L13Rik（简称 L13Rik）对高糖（high glucose, HG）培养的肾小球系膜细胞（mesangial cell, MC）损伤的影响及分子机制。
方法荧光定量PCR（quantitative real-time polymerase chain reaction, qRT-PCR）检测L13Rik和miR-2861在HG培养的肾小球MC中的表达水平；设计合成 L13Rik siRNA （siL13Rik）转染细胞，流式细胞术检测细胞周期，35S-甲硫氨酸标记实验评估细胞蛋白质合成，蛋白免疫印迹检测MC合成细胞外基质能力；双荧光素酶报告实验、RNA pull-down及RNA免疫沉淀实验评估L13Rik和miR-2861的靶向关系。
结果 HG诱导的MC中 L13Rik水平显著增加（P<0.001），而 miR-2861 水平降低（P<0.001）。L13Rik 消耗和 miR-2861过表达都有效地降低了 HG 诱导的 MC 损伤和细胞外基质集聚（P<0.001）。L13Rik 靶向并负调控miR-2861表达。
结论抑制L13Rik通过靶向miR-2861可减轻HG诱导的MC损伤。

Abstract:
Objective To explore the biological role of lncRNA-C920021L13Rik (L13Rik for short) in regulating high glucose (HG)-induced injury of glomerular mesangial cell (MC).
Methods The expressions of L13Rik and miR-2861 in HG-induced glomerular MC were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). And the function of L13Rik on regulating HG-induced MC injury and extracellular matrix (ECM) accumulation was examined by flow cytometry, 35S-methionine labeling protein synthesis assay and Western blot. A targeted conjugation of miR-2861 with L13Rik was verified by dual-luciferase reporter assay, RNA pull-down and RNA immunoprecipitation (RIP).

Results L13Rik level spiked markedly while miR-2861 level dropped in HG-treated MCs (P<0.001). Both L13Rik depletion and miR-2861 overexpression effectively lessened HG-induced cell injury and ECM accumulation. L13Rik functioned as a competing endogenous RNA (ceRNA) to sponge miR-2861 for accentuating MC injury.
Conclusion L13Rik promotes HG-induced MC injury through regulating miR-2861.

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