%0 Journal Article
%T miR-384-5p通过阻断BMP7转录后调控促进肾纤维化<br>miR-384-5p promotes renal fibrosis through blocking post-transcriptional regulation of BMP7
%A 张文静
%A 孙吉平
%A 吕 佳
%A 李 燕
%A 耿瀛洲
%A 邵耀中
%A 尹爱萍
%A 路万虹
%J 西安交通大学学报(医学版)
%D 2019
%R 10.7652/jdyxb201906011
%X 摘要：目的 研究miR-384-5p是否通过调节肾小管上皮细胞BMP7转录而影响肾纤维化。方法 建立小鼠单侧输尿管阻塞模型(unilateral ureteral obstructive, UUO)，流式细胞仪荧光激活细胞分选(FACS)分离肾组织获取肾小管上皮细胞进行培养，通过脂质体转染表达miR-384-5p的质粒、表达反义miR-384-5p的质粒及空白质粒；静脉注射空质粒(n=10)或反义-miR-384-5p质粒(n=10)，14周后处死UUO模型小鼠，肾组织切片进行Masson三色染色评估肾纤维化程度；Western blot和实时定量PCR检测miRNA靶基因(miR-384-5p、miR-30、miR-92和miR-128)表达。结果 UUO模型14d时肾脏出现明显纤维化，肾组织BMP7 mRNA明显增加，但BMP7蛋白无明显增加。miR-384-5p没有影响肾小管上皮细胞中BMP7 mRNA表达，但是过表达miR-384-5p的肾小管上皮细胞中BMP7蛋白明显减少，过表达反义miR-384-5p的肾小管上皮细胞中BMP7蛋白明显增加。免疫组化和定量PCR检测结果显示，UUO小鼠14 d肾组织纤维化显著增加，而注射反义miR-384-5p组肾组织纤维化程度明显减轻。虽然反义-miR-384-5p处理后不影响BMP7 mRNA水平，但能明显增加肾脏组织BMP7蛋白表达，提示抑制miR-384-5p能减轻UUO大鼠的肾损伤。结论 miR-384-5p是调节肾损伤后纤维化机制的关键因子，靶向miR-384-5p有希望成为防治肾脏纤维化的新方法。<br>ABSTRACT: Objective To investigate whether microRNA-384-5p can interfere with renal fibrosis by regulating BMP7 transcription in renal tubular epithelial cells. Methods Unilateral ureteral obstructive (UUO) model was established, and fluorescence activated cell sorter (FACS) was used to isolate renal tubular epithelial cells for culture. Plasmid expressing miR-384-5p, antisense miR-384-5p, and blank plasmid were transfected by liposome and injected in UUO mice. Masson staining was used to detect renal fibrosis, and Western blot and real-time PCR were used to detect the expressions of target-genes of miRNA (miR-384-5p, miR-30, miR-92, and miR-128). Results There was obvious renal fibrosis at 14 day in UUO model group; the expression of BMP7 mRNA in the renal tissue increased obviously while the expression of BMP7 protein was not increased. Although miR-384-5p could not change BMP7 mRNA in the renal tubular epithelial cells, BMP7 protein in the renal tubular epithelial cells that overexpressed miR-384-5p decreased significantly, and BMP7 protein in these cells overexpressing antisense miR-384-5p increased significantly. Immunohistochemistry and quantitative PCR showed that renal tissue fibrosis in UUO mice increased significantly in 14 days, while antisense miR-384-5p could significantly reduce the renal tissue fibrosis. Although antisense-miR-384-5p treatment did not affect the level of BMP7 mRNA, it could significantly increase the expression of BMP7 protein in renal tissue, suggesting that inhibiting miR-384-5p could lessen renal injury in UUO mice. Conclusion As a key factor regulating the mechanism of renal fibrosis after renal injury, targeting miR-384-5p may become a new method to prevent and treat renal fibrosis
%K 骨形态发生蛋白-7(BMP7)
%K 肾小管上皮细胞
%K miR-384-5p
%K 肾纤维化<br>BMP7
%K renal tubular epithelial cell
%K miR-384-5p
%K renal fibrosis
%U http://yxxb.xjtu.edu.cn//oa/darticle.aspx?type=view&id=201906011