| 崔凯,张睿,栾仲秋.微 RNA-181a-5p介导 Sirt1/PPARγ信号通路促进肾病综合征大鼠细胞凋亡的机制研究[J].安徽医药,2024,28(9):1756-1761. |
| 微 RNA-181a-5p介导 Sirt1/PPARγ信号通路促进肾病综合征大鼠细胞凋亡的机制研究 |
| Mechanism of apoptosis in rats with nephrotic syndrome by miR-181a-5p through Sirt1/PPARγ signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2024.09.014 |
| 中文关键词: 肾病综合征 肾足细胞 微 RNA-181a-5p 沉寂信息调节因子 过氧化物酶体增殖物激活受体 γ 凋亡 阿霉素 大鼠， Sprague-Dawley |
| 英文关键词: Nephrotic syndrome Podocytes miR-181a-5p Sirt1 PPARγ Apoptosis Adriamycin Rats, Sprague-Dawley |
| 基金项目:黑龙江省自然科学基金联合引导项目( LH2022H073) |
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| 中文摘要: |
| 目的旨在探究微 RNA(miR)-181a-5p对肾病综合征大鼠细胞凋亡的调控机制。方法该研究起止年限为 2021年 1月至 2022年 12月。使用阿霉素( ADR)构建肾病综合征大鼠模型及体外肾病综合征损伤模型，转染 miR-181a-5p inhibitor/NC至体外细胞模型。使用生化分析仪分析血清肌酐、血尿素氮、血清白蛋白和尿蛋白，实时荧光定量逆转录聚合酶链反应( qRTPCR)检测肾组织及大鼠肾足细胞 miR-181a-5p表达水平，通过 TargetScan(http://www.targetscan.org/vert_72/)预测 miR-181a-5p的下游靶基因，双萤光素酶报告实验来验证 miR-181a-5p与沉默信息调节因子 1(Sirt1)靶向结合关系。使用 CCK-8试剂盒检测细胞增殖，流式细胞术检测细胞凋亡，通过蛋白质印迹法检测 Sirt1及过氧化物酶体增殖物激活受体 γ(PPARγ)蛋白水平。结果肾病综合征大鼠 miR-181a-5p相对表达水平为 3.50±0.30，敲低 miR-181a-5p，使得 Sirt1蛋白表达水平从 0.36±0.02上调至0.87±0.06，PPARγ蛋白表达水平从 0.26±0.02上调至 0.78±0.05。结论 miR-181a-5p通过抑制 Sirt1/PPARγ信号通路活性促进肾足细胞凋亡。 |
| 英文摘要: |
| Objective To explore the regulatory mechanism of microRNA (miR)-181a-5p on apoptosis in rats with nephrotic syndrome. Methods The starting and ending years of the study were from January 2021 to December 2022. A rat model of nephrotic syndromeand an in vitro nephrotic syndrome injury model were constructed using adriamycin (ADR), and miR-181a-5p inhibitor/NC was transfect-ed into the in vitro cell model. Serum creatinine, blood urea nitrogen, serum albumin and urinary protein were analyzed using a biochemi-cal analyzer, and miR-181a-5p expression levels were detected in renal tissues and glomerular podocytes of rats by real-time fluores- cence quantitative reverse transcription polymerase chain reaction (qRT-PCR). TargetScan (http://www.targetscan.org/vert_72/) was ad- opted to predict the downstream target genes of miR-181a-5p, and dual luciferase reporter assay to verify the target binding relationship between miR-181a-5p and silent information regulator 1 (Sirt1). Cell proliferation was detected using CCK-8 kit, apoptosis by flow cy-tometry, and Sirt1 and peroxisome proliferator activated receptor γ (PPARγ) protein levels by protein blotting.Results The relative ex- pression level of miR-181a-5p in nephrotic syndrome rats was 3.50±0.30. Knockdown of miR-181a-5p resulted in the up-regulation of Sirt1 protein expression level from 0.36±0.02 to 0.87±0.06, and the up-regulation of PPARγ protein expression level from 0.26±0.02 to 0.78±0.05.Conclusion miR-181a-5p promotes glomerular podocyte apoptosis by inhibiting Sirt1/PPARγ signaling pathway activity. |
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