miR-29c 通过 AMPK/mTOR 信号通路对糖尿病大鼠肾纤维化的影响。
Effect of miR-29c on renal fibrosis in diabetic rats via the AMPK/mTOR signaling pathway.
机构信息
Department of Nephrology, Jining No. 1 People's Hospital, Jining, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Jul;23(14):6250-6256. doi: 10.26355/eurrev_201907_18445.
OBJECTIVE
To explore the effect of micro-ribonucleic acid (miR)-29c on renal fibrosis in diabetic rats through the adenosine 5'-monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway, and to investigate its related mechanism by the research on the effect of miR-29c on the expression of fibrosis-related genes.
MATERIALS AND METHODS
The rat model of diabetic nephropathy (DN) was established, and the levels of fasting blood glucose (FBG), 24 h urine protein (24h-Pro), blood urea nitrogen (BUN), and serum creatinine (sCr) were monitored. After the rats were executed, kidney tissues were dissected, stained with paraffin and embedded in hematoxylin and eosin (H&E). Then, Western blotting was used to detect the levels of miR-29c, phosphorylated-AMPK (p-AMPK), α-smooth muscle actin (α-SMA), tumor necrosis factor-α (TNF-α), and macrophage migration inhibitory factor (MIF). The human renal tubular epithelial HK-2 cell line was treated with high glucose (HG) to simulate DN cell status in vivo. After that, the expressions of miR-29c and the renal fibrosis marker α-SMA were examined via Western blotting. Finally, the level of α-SMA was measured by Western blotting after HG treatment combined with miR-29c silencing.
RESULTS
The levels of FBG, BUN, sCr, and 24h-Pro in DN model rats were significantly higher than those in rats of control group. The data manifested that the DN model was successfully established. The expression level of miR-29c in DN model rats was markedly increased and that of the downstream protein p-AMPK also exhibited a significantly increasing trend. In addition, the levels of α-SMA, TNF-α, and MIF were elevated. The expression levels of miR-29c and α-SMA were increased markedly after the human renal tubular epithelial HK-2 cell line was treated with HG, but the expression of α-SMA was reduced after HG treatment combined with miR-29c silencing for 24 h.
CONCLUSIONS
MiR-29c is probably related to the occurrence and development of DN. Besides, miR-29c silencing may inhibit the DN renal fibrosis through AMPK/mTOR signals, so miR-29c may be an effective target for the treatment of DN renal fibrosis.
目的
通过腺苷 5'-单磷酸激活蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路探讨微小 RNA（miR）-29c 对糖尿病大鼠肾纤维化的影响，并通过研究 miR-29c 对纤维化相关基因表达的影响来探讨其相关机制。
材料与方法
建立糖尿病肾病（DN）大鼠模型，监测空腹血糖（FBG）、24 小时尿蛋白（24h-Pro）、血尿素氮（BUN）和血肌酐（sCr）水平。处死大鼠后，分离肾脏组织，进行石蜡包埋和苏木精-伊红（H&E）染色。然后，采用 Western blot 法检测 miR-29c、磷酸化 AMPK（p-AMPK）、α-平滑肌肌动蛋白（α-SMA）、肿瘤坏死因子-α（TNF-α）和巨噬细胞移动抑制因子（MIF）的水平。采用高糖（HG）处理人肾小管上皮细胞 HK-2 细胞系，模拟体内 DN 细胞状态。然后，通过 Western blot 法检测 miR-29c 和肾纤维化标志物α-SMA 的表达。最后，在 HG 处理结合 miR-29c 沉默后，通过 Western blot 法测量α-SMA 的水平。
结果
DN 模型大鼠的 FBG、BUN、sCr 和 24h-Pro 水平明显高于对照组大鼠。这表明 DN 模型成功建立。DN 模型大鼠 miR-29c 表达水平明显升高，下游蛋白 p-AMPK 也呈明显升高趋势。此外，α-SMA、TNF-α和 MIF 的水平也升高。人肾小管上皮细胞 HK-2 细胞系用 HG 处理后，miR-29c 和α-SMA 的表达水平明显升高，但 HG 处理 24 小时后结合 miR-29c 沉默，α-SMA 的表达水平降低。
结论
miR-29c 可能与 DN 的发生发展有关。此外，miR-29c 沉默可能通过 AMPK/mTOR 信号抑制 DN 肾纤维化，因此 miR-29c 可能是治疗 DN 肾纤维化的有效靶点。
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