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沉默附睾血管内皮生长因子(VEGF)表达通过下调 VEGF 受体 2(VEGFR2)对砷中毒大鼠透明质酸酶(HYD)活性的影响。

Effects of silencing epididymal vascular endothelial growth factor (VEGF) expression on hyaluronidase (HYD) activity in arsenic poisoning rats through downregulating VEGF receptor 2 (VEGFR2).

机构信息

Department of Pathology and Pathophysiology, College of Basic Medical Science, Guizhou Medical University, Guiyang, Guizhou, China.

Department of Internal Medicine, People's Hospital in Yueyanglou District, Yueyang, Hunan, China.

出版信息

Bioengineered. 2021 Dec;12(1):1351-1359. doi: 10.1080/21655979.2021.1915726.

Abstract

RNA interference (RNAi) was used to investigate the role of epididymal vascular endothelial growth factor (VEGF) gene expression on sperm hyaluronidase (HYD) in a rat model of arsenic poisoning and to identify a new gene therapy target for male infertility caused by arsenic poisoning. The Rat model of chronic arsenic poisoning was established. And we found that positive expression of VEGF and VEGF receptor 2 (VEGFR2) was observed by Immunohistochemical staining in the epididymal tissues of arsenic-exposed rats. Subsequently, VEGF-shRNA-1, VEGF-shRNA-2 and VEGF shRNA-3 expression vectors containing epididymal VEGF-shRNA lentivirus were constructed and injected into the bilateral epididymis of each group of rats (Control group, NC-shRNA negative infection group, VEGF-shRNA-1 group, VEGF-shRNA-2 group, VEGF-shRNA-3 group) (n = 10 per group). Compared with the negative infection group and the normal control group, the expression of VEGF and VEGFR2 mRNA and protein levels were significantly decreased following epididymal infection. In addition, the HYD activity was all significantly lower than that in the normal control group and the negative infection group. Taken together, epididymal VEGF gene silencing may inhibit the activity of sperm HYD through downregulating VEGFR2.

摘要

RNA 干扰(RNAi)被用于研究附睾血管内皮生长因子(VEGF)基因表达在砷中毒大鼠模型中对精子透明质酸酶(HYD)的作用,并确定砷中毒引起的男性不育的新基因治疗靶点。建立了慢性砷中毒大鼠模型。我们发现免疫组织化学染色显示,砷暴露大鼠的附睾组织中观察到 VEGF 和 VEGF 受体 2(VEGFR2)的阳性表达。随后,构建了包含附睾 VEGF-shRNA 慢病毒的 VEGF-shRNA-1、VEGF-shRNA-2 和 VEGF shRNA-3 表达载体,并将其注入每组大鼠(对照组、NC-shRNA 阴性感染组、VEGF-shRNA-1 组、VEGF-shRNA-2 组、VEGF-shRNA-3 组)(n=10/组)的双侧附睾中。与阴性感染组和正常对照组相比,附睾感染后 VEGF 和 VEGFR2 mRNA 和蛋白水平的表达明显降低。此外,HYD 活性均明显低于正常对照组和阴性感染组。综上所述,附睾 VEGF 基因沉默可能通过下调 VEGFR2 抑制精子 HYD 的活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e44d/8806240/7bb26dd6055e/KBIE_A_1915726_UF0001_OC.jpg

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