Effects of HIF-1α on Spermatogenesis of Varicocele Rats by Regulating VEGF/PI3K/Akt Signaling Pathway
Effects of HIF-1α on Spermatogenesis of Varicocele Rats by Regulating VEGF/PI3K/Akt Signaling Pathway
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DOI：
10.1007/s43032-020-00395-0
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发表时间：
2020-11-25
影响因子：
2.9
通讯作者：
Qin，Qin
中科院分区：
文献类型：
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作者：
Wang，Danfeng;Zhao，Wei;Qin，Qin
缺氧诱导因子-1α (HIF-1α)参与了精索静脉曲张（VC）大鼠生精功能的调控，PI3K/Akt通路在其中发挥了重要作用。本研究采用CRISPR/Cas9基因编辑技术，沉默VC大鼠睾丸HIF-1α基因，探讨HIF-1α通过PI3K/Akt通路对VC大鼠生精细胞凋亡的影响。将sd大鼠随机分为正常大鼠组（N组）、VC模型组（V组）、VC + hif -1α-慢病毒组（H组）、VC +荧光素酶-慢病毒组（L组）。采用TUNEL试剂盒检测大鼠睾丸生精细胞凋亡情况。光镜下观察精小管的形态变化。采用Western blot、免疫荧光、免疫组化等方法检测VEGF、Akt、p-Akt、p70S6K、p-p70S6K的表达。采用单因素方差分析分析组间差异。与N组相比，V组生殖细胞分布紊乱，生精细胞凋亡明显增加，VEGF、p-Akt、p-p70S6K的表达也增加。与V组相比，H组精系上皮损伤改善，精系上皮排列基本有序。生精细胞凋亡明显减少，VEGF、P - akt、P - p70s6k蛋白表达降低（P< 0.05）。N组与H组间差异无统计学意义（P < 0.05）。综上所述，精索静脉曲张大鼠缺氧可调节HIF-1α调控其下游基因VEGF，而PI3K/Akt信号通路在这一过程中起调控作用。
Hypoxia-inducible factor-1α (HIF-1α) participates in the regulation of spermatogenic function in rats with varicocele (VC), and the PI3K/Akt pathway plays an important role in it. In the present research, we applied the CRISPR/Cas9 gene editing technique to silence the HIF-1α gene of VC rat testis, to explore the effect of HIF-1α on apoptosis of spermatogenic cells in VC rats through the PI3K/Akt pathway. Sprague Dawley rats were randomly assigned to four groups, including the normal rat group (group N), VC model group (group V), VC + HIF-1α-lentivirus group (group H), and VC + luciferase-lentivirus group (group L). Apoptosis of spermatogenic cells in rat testis was tested by TUNEL Kit. The morphologic changes of seminiferous tubules were viewed by a light microscope. Expressions of VEGF, Akt, p-Akt, p70S6K, and p-p70S6K were detected by means of Western blot, immunofluorescence, or immunohistochemistry methods. One-way ANOVA was applied to analyze the diverseness between groups. Compared with group N, the distribution of germ cells was disordered, apoptosis of spermatogenic cells increased significantly, and the expression of VEGF, p-Akt, and p-p70S6K was also increased in group V. Compared with group V, the damage of seminiferous epithelium in group H was improved, and the arrangement of the seminiferous epithelium was almost orderly. Apoptosis of spermatogenic cells decreased significantly, and the expression of VEGF, p-Akt, and p-p70S6K protein was decreased (P< 0.05). There was no significant difference between group N and group H (P> 0.05).In conclusion, HIF-1α is regulated by hypoxia in rats with varicocele to regulate its downstream gene VEGF which regulates spermatogenesis, and the PI3K/Akt signaling pathway plays a regulatory role in this process.