HIF1A and VEGF regulate each other by competing endogenous RNA mechanism and involve in the pathogenesis of peritoneal fibrosis
HIF1A and VEGF regulate each other by competing endogenous RNA mechanism and involve in the pathogenesis of peritoneal fibrosis
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HIF1A与VEGF通过竞争内源性RNA机制相互调节参与腹膜纤维化的发病机制
DOI：
10.1016/j.prp.2018.12.022
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发表时间：
2019-04-01
影响因子：
2.8
通讯作者：
Guo, Zhi Yong
中科院分区：
文献类型：
--
作者：
Li, Juan;Li, Shuang Xi;Guo, Zhi Yong
背景：腹膜纤维化是长期腹膜透析的主要难治性并发症，最终会导致腹膜超滤失败和腹膜透析终止。据报道，缺氧诱导因子1- α (HIF1A)调节血管内皮生长因子（VEGF）并参与腹膜纤维化，但其确切的分子调控机制尚不清楚。方法：采用酶联免疫吸附法测定42例腹膜患者的HIF1A和VEGF蛋白水平。通过生物信息学、逆转录聚合酶链反应、相关分析、RNA干扰、基因过表达和荧光素酶分析，阐明HIF1A和VEGF之间的竞争内源RNA （ceRNA）调控。结果：超滤问题腹膜透析患者腹膜流出液中HIF1A和VEGF水平均升高，且在蛋白水平和mRNA水平上呈正相关。生物信息学分析鉴定出8种常见的HIF1A和VEGF靶向mirna，包括miR-17-5p、20a、20b、93、106a、106b、199a-5p和203。MiR-17-5p被证明存在于患者的腹膜流出物中，并被选中进行进一步的研究。HIF1A mRNA和VEGF mRNA可以相互调控，调控需要miR-17-5p参与。HIF1A mRNA和VEGF mRNA下调/上调导致miR-17-5p上调/下调。此外，miR-17-5p的下调/上调与HIF1A mRNA和VEGF mRNA的上调/下调相关。荧光素酶检测表明HIF1A和VEGF通过3'UTR相互调节。结论：HIF1A和VEGF在腹膜间皮细胞中通过miR-17-5p和3'UTR介导相互调控，提示HIF1A和VEGF可能通过内源性RNA竞争机制相互调控腹膜纤维化的发生。
Background: Peritoneal fibrosis is a major intractable complication of long-term peritoneal dialysis, and would eventually lead to peritoneal ultrafiltration failure and the termination of peritoneal dialysis. Hypoxia-inducible factor 1-alpha (HIF1A) has been reported to regulate vascular endothelial growth factor (VEGF) and involves in peritoneal fibrosis, but the exact molecular regulation mechanism remains unknown.Methods: HIF1A and VEGF protein levels were measured in 42 peritoneal patients using enzyme linked immunosorbent assay. Bioinformatics, reverse transcription-polymerase chain reaction, correlation analysis, RNA interference, gene over-expression and luciferase assays were performed to clarify the competing endogenous RNA (ceRNA) regulation between HIF1A and VEGF.Results: Both HIF1A and VEGF levels were elevated in the peritoneal effluent of peritoneal dialysis patients with ultrafiltration problems, and were positively correlated with each other at protein level and mRNA level. Bioinformatics analysis identified 8 common targeted miRNAs for HIF1A and VEGF, including miR-17-5p, 20a, 20b, 93, 106a, 106b, 199a-5p and 203. MiR-17-5p was proved to be present in patients' peritoneal effluent and selected for further studies. HIF1A mRNA and VEGF mRNA could regulate each other, and miR-17-5p was required in the regulations. Down/up regulation of HIF1A mRNA and VEGF mRNA resulted in up/down regulation of miR-17-5p. Furthermore, down/up regulation of miR-17-5p was associated with up/down regulation of HIF1A mRNA and VEGF mRNA. Luciferase assay indicated that HIF1A and VEGF regulated each other through 3'UTR.Conclusion: HIF1A and VEGF could regulate each other in peritoneal mesothelial cell in the mediation of miR-17-5p and 3'UTR, indicating HIF1A and VEGF might regulate each other through competing endogenous RNA mechanism in the development of peritoneal fibrosis.