| 宋现静,刘明,吕文山.miR-103靶向钙调磷酸酶调节蛋白 1对糖尿病肾病足细胞损伤的影响[J].安徽医药,2021,25(10):1966-1971. |
| miR-103靶向钙调磷酸酶调节蛋白 1对糖尿病肾病足细胞损伤的影响 |
| miR-103 affects diabetic nephropathy podocyte injury by targeting RCAN1 gene |
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| DOI：10.3969/j.issn.1009-6469.2021.10.014 |
| 中文关键词: 糖尿病肾病 足细胞 微小 RNA-103 钙调磷酸酶调节蛋白 1(RCAN1) |
| 英文关键词: Diabetic nephropathy Podocyte miR-103 Regulator of calcineurin 1(RCAN1) |
| 基金项目:山东省自然科学基金( ZR2017MH069) |
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| 中文摘要: |
| 目的探讨微小 RNA-103(miR-103)在高糖诱导足细胞损伤中的作用及其可能的分子机制。方法本研究从 2018年 1月至2019年1月期间，体外培养人足细胞(HPC)以高糖刺激足细胞 6、12、24 h，qRT-PCR与Western blotting分别检测高糖刺激下 HPC细胞 miR-103、钙调磷酸酶调节蛋白 1(RCAN1)表，达；采用 ELISA检测白细胞介素 6(IL-6)、肿瘤坏死因子 -α(TNF-α)含量；流式细胞仪检测细胞凋亡率。检测干扰 miR-103的表达及 RCAN1过表达后细胞凋亡率及 IL-6、TNF-α水平变化；双荧光素酶报告基因检测验证miR-103与RCAN1的靶向调控关系。结果高糖诱导的足细胞中 miR-103的表达量升高［(1.00±0.07)比(4.51±0.34)］(P<0.05)，而 RCAN1表达水平降低［mRNA(1.00±0.08)(0.59±0.06)；蛋白(0.86±0.05)(0.36±0.04)］(P<0.05)，足细胞凋亡率增加［(5.31±0.52)%比( 24.36±1.58)%］(P<0.05)IL-6、 (18.84±0.85)ng/mL；(0.34±0.03)ng/mL比( 0.95±TNF-比α水平升高［(7.23±0.33)ng/mL比比0.07)ng/mL］(P<0.05)；干扰 miR-103表达及 RCAN1过表达后，足细胞凋亡率降低的，［( 22.87±1.55)%比( 14.72±0.82)%；(22.76±1.14)%比( 13.88±0.85)%］(P<0.05)IL-6［( 18.54±0.62)ng/mL比( 12.74±0.44)ng/mL；(19.74±0.61)ng/mL比( 14.87±0.56)ng/mL］、 TNF-α［(0.92±0.05)ng/mL比(0.54±0)ng/mL；(0.90±0.06)ng/mL比(0.56±0.04)ng/mL］水平降低(P<0.05)；miR-103与RCAN1存在.06， |
| 英文摘要: |
| Objective To explore the role of microRNA-103 (miR-103) in high glucose-induced podocyte injury and its possible molecular mechanism.Methods From January 2018 to January 2019 in this study, human foot cells (HPC) were cultured in vitro, and thepodocytes were stimulated with high glucose for 6, 12, and 24 h, respectively. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expressions of high-glucose-stimulated HPC cell miR-103, calcineurin regulatory protein 1 gene (RCAN1). The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). Apoptosis rate was measured by flow cytometry. The interfering expression of miR-103 was detected and the apoptosis rate and the expression levels of IL-6 and TNF-αwere measured after RCAN1 overexpression. Dual luciferase reporter gene assay verified the targeted regulation of miR-103 and RCAN1.Results The expression of miR-103 in podocytes induced by high glucose was increased [(1.00±0.07) vs. (4.51±0.34)] (P<0.05), while the expression level of RCAN1 was decreased [mRNA (1.00±0.08) vs. (0.59±0.06); protein (0.86±0.05) vs. (0.36±0.04)] (P<0.05), podocyte apoptosis rate was increased [(5.31±0.52)% vs. (24.36±1.58)%] (P<0.05), and the levels of IL-6, TNF-α were increased [(7.23±0.33) ng/mL vs. (18.84±0.85) ng/mL; (0.34±0.03) ng/mL vs. (0.95±0.07) ng/mL] (P<0.05). After interference with the expression of miR-103 and overexpression of RCAN1, the apoptosis rate of podocytes was decreased [(22.87±1.55)% vs. (14.72±0.82)%; (22.76±1.14)% vs. (13.88±0.85)%] (P<0.05), IL-6 [(18.54±0.62) ng/mL vs. (12.74±0.44)ng/mL; (19.74±0.61) ng/mL vs. (14.87±0.56)ng/mL], TNF-α[(0.92±0.05) ) ng/mL vs. (0.54±0.06)ng/mL; (0.90±0.06) ng/mL vs. (0.56±0.04)ng/mL] levels were decreased (P<0.05). The dual luciferase reporter gene assay demonstrated that miR-103 had a targeting relationship with RCAN1, and miR-103 could negatively regulate the expression and activity of RCAN1. Interference with the expression of RCAN1 partially reversed the protective effect of miR-103 expression on high glucose-stimulated podocytes.Conclusion Interfering with the expression of miR-103 can attenuate diabetic nephropathy podocyte injury by promoting the expression of RCAN1. |
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