miR- 34c- 5p targets ROCK1 expression to inhibit kidney injury in diabetic nephropathy rats through MAPK/ERK signaling pathway.

IF 1.5 4区生物学 Q4 CELL BIOLOGY

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-06-12 DOI:10.1007/s11626-025-01039-w

HuaJuan Wei, Ye Li, HongDe Liu, Li Pan, HuiLing Duo, ShaoYing Dong

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引用次数: 0

Abstract

This study was to investigate the mechanism of miR- 34c- 5p in alleviating kidney injury in diabetic nephropathy (DN) rats by targeting ROCK1 and MAPK/ERK signaling pathway. The rat model of DN was established and fasting blood glucose, 24-h proteinuria, blood urea nitrogen, and serum creatinine were measured to quantify kidney injury. Kidney tissue was dissected for H&E and TUNEL staining. Renal injury factor KIM- 1 was measured by Western blot analysis. Retinal Muller cells (RMCs) were treated with high glucose and transfected. Cell viability was detected by CCK- 8 and apoptosis by flow cytometry. Inflammatory factors in DN rats and RMCs were analyzed by ELISA. The targeting effect of miR- 34c- 54p on ROCK1 was demonstrated by RNA pull-down and dual-luciferase reporter gene. Finally, ROCK1, p-MEK1/2, and p-ERK were assessed by Western blot. Elevating miR- 34c- 5p could inhibit DN kidney injury and high glucose-induced cell injury, and reduce inflammation in kidney tissue of DN rats and RMCs. miR- 34c- 5p targeted to regulate ROCK1 expression, and restoring ROCK1 abolished the therapeutic effect of elevating miR- 34c- 5p. Phosphorylated MEK and ERK were increased in DN rats and RMCs induced by high glucose. miR- 34c- 5p can inhibit kidney injury induced by DN by targeting ROCK1, and the MAPK/ERK pathway may represent the pathological mechanism of DN.

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miR- 34c- 5p通过MAPK/ERK信号通路靶向ROCK1表达，抑制糖尿病肾病大鼠肾损伤。

本研究旨在探讨miR- 34c- 5p通过靶向ROCK1和MAPK/ERK信号通路减轻糖尿病肾病（DN）大鼠肾损伤的机制。建立DN大鼠模型，测定空腹血糖、24小时蛋白尿、尿素氮、血清肌酐，定量测定肾损伤。取肾组织进行H&E和TUNEL染色。Western blot法检测肾损伤因子KIM- 1。用高糖处理视网膜Muller细胞（RMCs）并转染。CCK- 8检测细胞活力，流式细胞术检测细胞凋亡。采用ELISA法分析DN大鼠和RMCs的炎症因子。miR- 34c- 54p对ROCK1的靶向作用通过RNA下拉和双荧光素酶报告基因得到证实。最后用Western blot检测ROCK1、p-MEK1/2和p-ERK。升高miR- 34c- 5p可抑制DN大鼠肾损伤和高糖诱导的细胞损伤，减轻DN大鼠和RMCs肾组织炎症反应。miR- 34c- 5p靶向调控ROCK1的表达，而ROCK1的恢复抵消了miR- 34c- 5p升高的治疗效果。高糖诱导的DN大鼠和RMCs中MEK和ERK磷酸化升高。miR- 34c- 5p可通过靶向ROCK1抑制DN所致肾损伤，MAPK/ERK通路可能代表DN的病理机制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

In Vitro Cellular & Developmental Biology. Animal 生物-发育生物学

CiteScore

3.70

自引率

4.80%

发文量

审稿时长

3 months

期刊介绍： In Vitro Cellular & Developmental Biology - Animal is a journal of the Society for In Vitro Biology (SIVB). Original manuscripts reporting results of research in cellular, molecular, and developmental biology that employ or are relevant to organs, tissue, tumors, and cells in vitro will be considered for publication. Topics covered include: Biotechnology; Cell and Tissue Models; Cell Growth/Differentiation/Apoptosis; Cellular Pathology/Virology; Cytokines/Growth Factors/Adhesion Factors; Establishment of Cell Lines; Signal Transduction; Stem Cells; Toxicology/Chemical Carcinogenesis; Product Applications.