缺氧诱导因子-1α调控VEGF/PI3K/Akt通路对缺氧大鼠睾丸支持细胞凋亡的影响及其机制  

Effect of hypoxia inducible factor⁃1αregulating VEGF/PI3K/Akt pathway on apoptosis of testicular Sertoli cells in hypoxic rats and its mechanism

作　　者：杨习曼 栗璐芳 张芳[2] 卢先锋[2] 刘建荣[2] 秦琴[2] YANG Ximan;LI Lufang;ZHANG Fang;LU Xianfeng;LIU Jianrong;QIN Qin(Department of Biochemistry and Molecular Biology,College of Basic Medicine,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China;不详)

机构地区：[1]山西医科大学基础医学院生物化学与分子生物学教研室,山西太原030001 [2]山西省人民医院,山西太原030012

出　　处：《中国生物制品学杂志》2024年第10期1167-1172,共6页Chinese Journal of Biologicals

基　　金：山西省基础研究计划项目(20210302123348);山西省卫生健康委“四个一批”科技兴医创新计划重点攻关专项(2022XM25)。

摘　　要：目的观察缺氧对支持细胞的影响,探讨缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)通过血管内皮生长因子(vascular endothelial growth factor,VEGF)/PI3K/Akt通路对缺氧条件下大鼠睾丸支持细胞凋亡的作用机制。方法分离、培养大鼠睾丸支持细胞,将细胞分为4组:C(正常培养不做任何处理)、V(缺氧培养48 h)、H(感染腺病毒敲低HIF-1α+缺氧培养48 h)、NC(感染空病毒载体+缺氧培养48 h)组,Western blot法检测HIF-1α、VEGF、p-Akt、Akt、p-p70S6K、p70S6K、Bax、Bcl-2蛋白的表达,免疫荧光法检测Bax蛋白在细胞中的表达和定位,流式细胞术检测细胞凋亡率。结果Western blot结果显示,与C、H组比较,V和NC组HIF-1α、VEGF、p-Akt、p-p70S6K、Bax蛋白的表达明显升高(F分别为11.71、20.34、10.28、6.096和18.55,P均<0.05),Bcl-2蛋白的表达明显降低(F=12.92,P<0.05);4组Akt、p70S6K蛋白的表达差异均无统计学意义(F分别为0.5727和0.1569,P均>0.05)。免疫荧光结果显示,Bax主要在胞质中表达,与C、H组比较,V和NC组Bax蛋白的表达明显升高(F=64.83,P<0.05)。流式细胞术结果显示,与C、H组比较,V和NC组凋亡率明显升高(F=9.649,P<0.05)。结论缺氧导致支持细胞凋亡可能是由于HIF-1α表达升高激活下游VEGF/PI3K/Akt信号通路所致。Objective To observe the effect of hypoxia on Sertoli cells,and explore the mechanism of hypoxia inducible factor-1α(HIF-1α)on apoptosis of Sertoli cells in testes of rats under hypoxia through VEGF/PI3K/Akt pathway.Methods Sertoli cells of rat testes were isolated and cultured,and the cells were divided into four groups:group C(normal culture without any treatment);group V(hypoxia culture for 48 h);group H(infected with adenovirus silencing HIF-1α+hypoxia culture for 48 h);NC group(infected with empty adenovirus vector+hypoxia culture for 48 h).The protein expressions of HIF-1α,VEGF,p-Akt,Akt,p-p70S6K,p70S6K,Bax and Bcl-2 were detected by Western blot.The expression and localization of Bax protein in cells were measured by immunofluorescence assay.The apoptosis rate was analyzed by flow cytometry.Results Western blot results showed that compared with group C and H,the protein expressions of HIF-1α,VEGF,p-Akt,pp70S6K and Bax in group V and NC significantly increased(F=11.71,20.34,10.28,6.096 and 18.55,respectively,each P<0.05),the protein expression of Bcl-2 significantly decreased(F=12.92,P<0.05).The expression of Akt and p70S6K proteins showed no significant difference among the four groups(F=0.5727 and 0.1569,respectively,each P>0.05).Immunofluorescence results showed that Bax was mainly expressed in the cytoplasm.Compared with group C and H,the expression of Bax protein in group V and NC significantly increased(F=64.83,P<0.05).The results of flow cytometry showed that compared with group C and H,the apoptosis rates of group V and NC significantly increased(F=9.649,P<0.05).Conclusion Hypoxia-induced apoptosis of Sertoli cells may be caused by the activation of VEGF/PI3K/Akt signaling pathway by the increased expression of HIF-1α.

关 键 词：缺氧诱导因子-1Α 支持细胞 VEGF/PI3K/Akt通路 细胞凋亡 

分 类 号：R34[医药卫生—基础医学]