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Mol Med. 2021 Jul 8;27(1):74. doi: 10.1186/s10020-021-00339-7.
BACKGROUND: Diabetic nephropathy (DN) is presently the main reason behind end-stage renal illness globally. The endothelial-to-mesenchymal transition (EndMT) of glomerular endothelial cells has been reported to play an important position in DN. As a particular type of epithelial-to-mesenchymal transition, EndMT and epithelial-to-mesenchymal transition could exhibit mutual modulators. Profilin 2 (PFN2) has been reported to take part in epithelial-to-mesenchymal transition. Furthermore, ETS proto-oncogene 1 (ets1) and lysine methyltransferase 5A (KMT5A) have been reported to contribute to excessive glucose-mediated endothelial damage and epithelial-to-mesenchymal transition. On this research, we hypothesize ets1 associates with KMT5A to modulate PFN2 transcription, thus collaborating in excessive glucose-mediated EndMT in glomerular endothelial cells.
METHODS: Immunohistochemistry (IHC) was carried out to detect protein ranges within the kidney tissues and/or aorta tissues of human topics and rats. Western blot, qPCR and immunofluorescence have been carried out utilizing human umbilical vein endothelial cells (HUVECs). Chromatin immunoprecipitation (ChIP) assays and twin luciferase assays have been carried out to evaluate transcriptional exercise. The distinction between the teams was in contrast by two-tailed unpaired t-tests or one-way ANOVAs.
RESULTS: Our knowledge indicated that vimentin, αSMA, S100A4 and PFN2 ranges have been elevated, and CD31 ranges have been diminished in glomerular endothelial cells of DN sufferers and rats. Our cell experiments confirmed that top glucose induced EndMT by augmenting PFN2 expression in HUVECs. Furthermore, excessive glucose elevated ets1 expression. si-ets1 suppressed excessive glucose-induced PFN2 ranges and EndMT. ets1 overexpression-mediated EndMT was reversed by si-PFN2. Moreover, ets1 was decided to affiliate with KMT5A. Excessive glucose attenuated KMT5A ranges and histone H4 lysine 20 methylation (H4K20me1), one of many downstream targets of KMT5A. KMT5A upregulation suppressed excessive glucose-induced PFN2 ranges and EndMT. sh-KMT5A-mediated EndMT was counteracted by si-PFN2. Moreover, H4K20me1 and ets1 occupied the PFN2 promoter area. sh-KMT5A cooperated with ets1 overexpression to activate PFN2 promoter exercise. Our in vivo research demonstrated that KMT5A was diminished, whereas ets1 was augmented, in glomerular endothelial cells of DN sufferers and rats.
CONCLUSIONS: The current research indicated that ets1 cooperated with KMT5A to transcribe PFN2, thus contributing to hyperglycemia-induced EndMT within the glomerular endothelial cells of DN sufferers and rats. Trial registration ChiCTR, ChiCTR2000029425. 2020/1/31, http://www.chictr.org.cn/showproj.aspx?proj=48548.