Xing Chenhao, Lu Yamin, Liu Guangxia, Chen Fang, Hou Zhan, Zhang Yiwen
Graduate School of Hebei North University, Hebei North University, Zhangjiakou, Hebei, People's Republic of China.
Department of Nuclear Medicine, Hebei General Hospital, Shijiazhuang, Hebei, People's Republic of China.
Diabetes Metab Syndr Obes. 2024 Feb 23;17:903-911. doi: 10.2147/DMSO.S449791. eCollection 2024.
The purpose of the study was to investigate the expression levels and correlation of inflammatory factors such as miR-377-3p and TGF-β in patients with diabetic kidney disease (DKD), and to investigate the regulatory mechanism of transfection of miR-377-3p on the inflammatory response of HK-2 cell induced by high glucose.
According to UACR, patients were divided into normal albuminuria group (Con, n = 29), microalbuminuria group (Micro, n = 31) and macroalbuminuria group (Macro, n = 30), analyzed the correlation and influencing factors between DKD and inflammatory factor. HK-2 cells were randomly divided into four groups: normal control group (NC), high glucose group (HG), miR-377-3p overexpression group (MIN), and miR-377-3p inhibition group (IN). After transfection of miR-377-3p mimics and inhibitors, the contents of TGF-β, IL-6 and IL-18 were detected by RT-PCR and Western blot.
The levels of miR-377-3p, TGF-β, IL-6 and IL-18 in both Micro group and Macro group were significantly higher than those in Con group (P < 0.05); Pearson correlation analysis showed that miR-377-3p was positively correlated with UACR, TG, TGF-β, IL-6 and IL-18, and negatively correlated with GFR (P < 0.05). Cell experiment: RT-PCR and Western blot results showed that miR-377-3p, TGF-β, IL-6 and IL-18 in HG group were significantly higher than those in NC group (P < 0.05). After transfection with miR-377-3p inhibitor, the levels of miR-377-3p, TGF-β, IL-6 and IL-18 in IN group were significantly decreased compared with HG group and MIN group.
miR-377-3p expression was elevated both in serum of DKD patients and in HK-2 cells with high glucose induced injury, overexpression of miR-377-3p exacerbates the damage to HK-2 cells and promotes the progression of DKD. Silencing miR-377-3p can potentially regulate the levels of inflammatory factors in HK-2 cells by targeting downregulation of TGF-β expression, thereby mitigating the damage to HK-2 cells and delaying the development of diabetic kidney disease.
本研究旨在探讨糖尿病肾病（DKD）患者中miR-377-3p和转化生长因子-β（TGF-β）等炎症因子的表达水平及其相关性，并研究miR-377-3p转染对高糖诱导的HK-2细胞炎症反应的调控机制。
根据尿白蛋白肌酐比值（UACR），将患者分为正常蛋白尿组（Con，n = 29）、微量白蛋白尿组（Micro，n = 31）和大量白蛋白尿组（Macro，n = 30），分析DKD与炎症因子之间的相关性及影响因素。将HK-2细胞随机分为四组：正常对照组（NC）、高糖组（HG）、miR-377-3p过表达组（MIN）和miR-377-3p抑制组（IN）。转染miR-377-3p模拟物和抑制剂后，通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测TGF-β、白细胞介素-6（IL-6）和白细胞介素-18（IL-18）的含量。
Micro组和Macro组中miR-377-3p、TGF-β、IL-6和IL-18水平均显著高于Con组（P < 0.05）；Pearson相关性分析显示，miR-377-3p与UACR、甘油三酯（TG）、TGF-β、IL-6和IL-18呈正相关，与肾小球滤过率（GFR）呈负相关（P < 0.05）。细胞实验：RT-PCR和蛋白质免疫印迹结果显示，HG组中miR-377-3p、TGF-β、IL-6和IL-18水平显著高于NC组（P < 0.05）。用miR-377-3p抑制剂转染后，IN组中miR-377-3p、TGF-β、IL-6和IL-18水平与HG组和MIN组相比显著降低。
DKD患者血清及高糖诱导损伤的HK-2细胞中miR-377-3p表达均升高，miR-377-3p过表达加剧HK-2细胞损伤并促进DKD进展。沉默miR-377-3p可通过靶向下调TGF-β表达潜在调节HK-2细胞中炎症因子水平，从而减轻HK-2细胞损伤并延缓糖尿病肾病发展。
