Zhu Pei, Qi Tao, Huang Zhan-Sen, Li Hao, Wang Bo, Feng Jia-Xin, Ma Shuai, Xiao Heng-Jun, Tang Yu-Xin, Liu Wei, Chen Jun
Department of Infertility and Sexual Medicine, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China.
Department of Urology, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China.
Transl Androl Urol. 2020 Apr;9(2):218-231. doi: 10.21037/tau.2020.03.11.
Erectile dysfunction (ED) is a well-known complication of diabetes, affecting up to 75% of diabetic men. Although the etiology of diabetic ED is multifactorial, endothelial dysfunction is considered to be a pillar of its pathophysiology. Endothelial dysfunction is caused by the harmful effects of high glucose levels and increased oxidative stress on the endothelial cells that comprise the vascular endothelium. The aim of this study was to identify the proteomic changes caused by high glucose-induced oxidative stress and explore the role of heme oxygenase 1 (HMOX1) in it.
The cellular proteomic response to hypoxanthine-induced oxidative stress in human umbilical vein endothelial cells (HUVECs) was analyzed by isobaric tags for relative and absolute quantitation (iTRAQ) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Differentially expressed proteins (DEPs) were analyzed through Network and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Further validation assays was performed to validate the role of HMOX1.
The results showed that 66 and 76 DEPs were markedly upregulated and downregulated, respectively, for HUVECs oxidative stress. Among these proteins, we verified eight dysregulated genes by quantitative reverse transcription PCR, including nucleolin (NCL), X-ray repair cross-complementing protein 6 (XRCC6), ubiquinol-cytochrome C reductase binding protein (UQCRB), non-POU domain containing octamer binding (NONO), heme oxygenase 1 (HMOX1), nucleobindin 1 (NUCB1), DEK, and chromatin target of prmt1 (CHTOP). Further, using overexpression and genetic knockdown approaches, we found that HMOX1 was critical for the oxidative stress response in HUVECs.
We found that HMOX1 was closely related to the oxidative stress response induced by hypoxanthine. To the best of our knowledge, this study is the first overview of the responses of HUVECs to oxidative stress. The findings will contribute to analyses of the detailed molecular mechanisms involved in the pathogenesis of endothelial dysfunction and related molecular mechanisms in ED patients.
勃起功能障碍（ED）是糖尿病一种众所周知的并发症，影响高达75%的糖尿病男性。尽管糖尿病性ED的病因是多因素的，但内皮功能障碍被认为是其病理生理学的一个支柱。内皮功能障碍是由高血糖水平和氧化应激增加对构成血管内皮的内皮细胞的有害影响所引起的。本研究的目的是确定高糖诱导的氧化应激所导致的蛋白质组学变化，并探讨血红素加氧酶1（HMOX1）在其中的作用。
采用相对和绝对定量的等压标签（iTRAQ）结合液相色谱 - 串联质谱（LC - MS/MS）分析人脐静脉内皮细胞（HUVECs）对次黄嘌呤诱导的氧化应激的细胞蛋白质组学反应。通过网络分析和京都基因与基因组百科全书（KEGG）通路分析对差异表达蛋白（DEPs）进行分析。进行进一步的验证试验以验证HMOX1的作用。
结果显示，对于HUVECs氧化应激，分别有66个和76个DEPs显著上调和下调。在这些蛋白质中，我们通过定量逆转录PCR验证了8个失调基因，包括核仁素（NCL）、X射线修复交叉互补蛋白6（XRCC6）、泛醌 - 细胞色素C还原酶结合蛋白（UQCRB）、含非POU结构域的八聚体结合蛋白（NONO）、血红素加氧酶1（HMOX1）、核结合蛋白1（NUCB1）、DEK和蛋白质精氨酸甲基转移酶1的染色质靶点（CHTOP）。此外，使用过表达和基因敲低方法，我们发现HMOX1对HUVECs中的氧化应激反应至关重要。
我们发现HMOX1与次黄嘌呤诱导的氧化应激反应密切相关。据我们所知，本研究是对HUVECs对氧化应激反应的首次概述。这些发现将有助于分析内皮功能障碍发病机制中涉及的详细分子机制以及ED患者的相关分子机制。
