miRNA-152在损伤足细胞中异常表达的研究  

Study on the abnormal expression of miRNA-152 in injured podocytes

作　　者：张宏波 苏彩霞 沈晓丹 张学明[1] Zhang Hongbo;Su Caixia;Shen Xiaodan;Zhang Xueming(Department of Biochemistry and Molecular Biology,Inner Mongolia University of Science and Technology Baotou Medical College,Baotou,Inner Mongolia,014060,China)

机构地区：[1]内蒙古科技大学包头医学院生物化学与分子生物学教研室,内蒙古包头014060

出　　处：《当代医学》2020年第21期1-3,共3页Contemporary Medicine

基　　金：内蒙古自治区科技创新引导项目(2019);内蒙古自治区自然科学基金项目(2016MS(LH)0308);内蒙古自治区高等学校科学研究项目(NJZY16215)。

摘　　要：目的以阿霉素(Adriamycin,ADR)诱导的足细胞损伤模型,来研究miRNA-152在损伤足细胞中的表达,为阐明miRNA-152异常表达导致肾足细胞损伤的机制研究奠定基础。方法将0.00μg/mL,0.05μg/mL,0.10μg/mL,0.20μg/mL,0.40μg/mL,0.80μg/mL浓度的ADR加入已分化培养6 d的足细胞中,诱导培养24 h后,通过足细胞骨架免疫荧光染色和Synaptopodin表达检测,鉴定足细胞的损伤;然后采用Real-time PCR法检测miRNA-152和14-3-3β的表达,分析miRNA-152和14-3-3β的表达关系。结果分化培养6 d的足细胞在加入ADR 24 h后,鬼笔环肽免疫荧光染色结果显示,ADR诱导培养的足细胞中肌动蛋白应力纤维减少,且细胞骨架排列失调;Real-time PCR检测结果表明,ADR诱导培养的足细胞中,Synaptopodin的表达下降。在损伤足细胞中,Real-time PCR法检测结果表明,miRNA-152的表达上调,14-3-3β的表达则下降。结论在损伤足细胞中,miRNA-152表达上调,而14-3-3β的表达则下降,提示了miRNA-152可能直接下调了14-3-3β的表达,进而导致足细胞损伤。Objective We aimed to study the expression of miRNA-152 and find out the correlation between miRNA-152 and 14-3-3β expression in proteinuria nephropathy by a model of ADR-induced podocyte injury. Methods ADR at a concentration of 0.00 μg/mL, 0.05 μg/mL, 0.10 μg/mL,0.20 μg/mL, 0.40 μg/mL, and 0.80 μg/mL was used to induce podocyte injury for 24 hours. The podocyte injury was assessed by using cytoskeleton immunofluorescence staining and detection of Synaptopodinexpression by Real-time PCR in the podocytes. The expression of miRNA-152 and 14-3-3β was then detected by Real-time PCR in the injuted podocytes. Results After the podocytes were induce to injury by ADR for 24 hours. The podocyte cytoskeleton was examined by means of fluorescein-conjugated phalloidin staining, and it was observed that the stress fibers in the podocytes were severely reducted and disordered. The results of Real-time PCR showed that the expression of Synaptopodin mRNA decreased in ADR-induced culture podocytes. In the injured podocytes, the results of Real-time PCR showed that the expression of miRNA-152 was up-regulated and the expression of 14-3-3β was decreased. Conclusion miRNA-152 expression was up-regulated and 14-3-3β expression was decreased in the injured podocytes, which suggested that miRNA-152 might directly down-regulate the expression of 14-3-3β and then lead to podocyte damage.

关 键 词：蛋白尿肾病 足细胞损伤 miRNA-152 14-3-3-β SYNAPTOPODIN 

分 类 号：R692[医药卫生—泌尿科学]