Han Xu, Li Qiaobei, Wang Chunyan, Li Yinyan
Department of Traditional Chinese medicine, the First Affiliated Hospital of China Medical University, Shenyang, China.
Department of Ultrasonic Diagnosis, the First Affiliated Hospital of China Medical University, Shenyang, China.
Exp Clin Endocrinol Diabetes. 2019 Jun;127(6):387-395. doi: 10.1055/a-0630-0173. Epub 2018 Jun 25.
Previous study has been reported that braykinin B2 receptor (Bdkrb2) involves in high glucose-induced renal and podocytes injuries. However, there have been some studies with contradictory results that Bdkrb2 has a protective effect on hyperglycemia-induced injuries in vivo and in vitro. The purpose of the present study was carried out to further investigate the post-transcriptional regulatory mechanism of microRNA (miR) in high glucose-treated podocytes by targeting Bdkrb2 signaling in vitro.
The CCK-8 and flow cytometry were performed to measure the cell viability and apoptosis. Gene and protein expression were assayed by RT-qPCR and western blotting, respectively.
High glucose treatment decreased cell viability and induced membrane and DNA damage, as well as apoptosis in podocytes. High glucose treatment also increased the expression of Bdkrb2, which was blocked by miR-204-3p mimics transfection in podocytes. Bioinformatics and luciferase reporter activity showed that miR-204-3p was directly targeted to the 3'-untranslated region (3'-UTR) of Bdkrb2. High glucose-induced apoptosis and dysfunction in podocytes were reserved by miR-204-3p mimics transfection, while the effects of miR-204-3p mimics in high glucose-treated podocytes were neutralized by overexpressed Bdkrb2.
These findings suggested that miR-204-3p may play a protective role in high glucose-induced apoptosis and dysfunction in podocytes through down-regulation of Bdkrb2.
先前的研究报道称，缓激肽B2受体（Bdkrb2）参与高糖诱导的肾脏和足细胞损伤。然而，也有一些研究结果相互矛盾，表明Bdkrb2在体内和体外对高血糖诱导的损伤具有保护作用。本研究的目的是在体外通过靶向Bdkrb2信号通路进一步探讨微小RNA（miR）在高糖处理的足细胞中的转录后调控机制。
采用CCK-8法和流式细胞术检测细胞活力和凋亡情况。分别通过RT-qPCR和蛋白质印迹法检测基因和蛋白表达。
高糖处理降低了足细胞的活力，诱导了细胞膜和DNA损伤以及细胞凋亡。高糖处理还增加了Bdkrb2的表达，而在足细胞中转染miR-204-3p模拟物可阻断这种增加。生物信息学和荧光素酶报告基因活性表明，miR-204-3p直接靶向Bdkrb2的3'-非翻译区（3'-UTR）。转染miR-204-3p模拟物可逆转高糖诱导的足细胞凋亡和功能障碍，而在高糖处理的足细胞中，miR-204-3p模拟物的作用被过表达的Bdkrb2中和。
这些发现表明，miR-204-3p可能通过下调Bdkrb2对高糖诱导的足细胞凋亡和功能障碍发挥保护作用。
