a Department of Urology, Henan Provincial People's Hospital , People's Hospital of Zhengzhou University , Zhengzhou , China.
Cell Cycle. 2019 Jan;18(2):119-129. doi: 10.1080/15384101.2018.1542900. Epub 2019 Jan 3.
In this study, we aimed to reveal the role of miR-191 in apoptosis of renal tubular epithelial cells and in the involvement of renal ischemia-reperfusion injury. Renal transplantation rat model was established. miR-191 and Cystathionine-β-synthase (CBS) were measured by qRT-PCR and Western blot. The regulation of miR-191 on CBS was detected by luciferase reporter assay. We found miR-191 expression in platelets and platelet microvesicles (P-MVs) of patients and model rats was significantly upregulated than that of health and normal rats. Also, mRNA and protein levels of CBS in renal tissues of patients were significantly downregulated than that of health and normal rats. We also found that P-MVs could transfer miR-191 to HK-2 cells. Luciferase reporter assay showed that CBS was a direct target of miR-191. In addition, we proved that P-MVs-secreted miR-191 inhibited CBS expression in HK-2 cells, and P-MVs-secreted miR-191 promoted HK-2 cell apoptosis via CBS. Finally, we verified the trends of CBS expressions, HK-2 cell apoptosis and apoptosis-related proteins in vivo were similar as the trends in vitro. Therefore, CBS was a direct target of miR-191, and miR-191 could transfer to HK-2 cells via P-MVs to decrease the expression of CBS, thus to promote cell apoptosis and renal IR injury.
在这项研究中,我们旨在揭示 miR-191 在肾小管上皮细胞凋亡和肾缺血再灌注损伤中的作用。建立了肾移植大鼠模型。通过 qRT-PCR 和 Western blot 检测 miR-191 和胱硫醚-β-合酶 (CBS)。通过荧光素酶报告基因检测 miR-191 对 CBS 的调控。我们发现患者和模型大鼠血小板和血小板微囊泡 (P-MV) 中的 miR-191 表达明显高于健康和正常大鼠。此外,患者肾组织中 CBS 的 mRNA 和蛋白水平明显低于健康和正常大鼠。我们还发现 P-MV 可以将 miR-191 转染到 HK-2 细胞中。荧光素酶报告基因检测显示 CBS 是 miR-191 的直接靶标。此外,我们证明 P-MV 分泌的 miR-191 抑制 HK-2 细胞中的 CBS 表达,并且 P-MV 分泌的 miR-191 通过 CBS 促进 HK-2 细胞凋亡。最后,我们验证了 CBS 表达、HK-2 细胞凋亡和凋亡相关蛋白的体内趋势与体外趋势相似。因此,CBS 是 miR-191 的直接靶标,miR-191 可以通过 P-MV 转移到 HK-2 细胞中,降低 CBS 的表达,从而促进细胞凋亡和肾 IR 损伤。