FKBP 5基因对绵羊卵泡颗粒细胞功能的影响

Effects of FKBP 5 on Function of Sheep Follicular Granulosa Cells

旨在探讨FKBP 5在绵羊卵巢颗粒细胞(GCs)中的潜在功能。本研究从乌鲁木齐市华凌屠宰场的20个2岁左右处于性成熟阶段的阿勒泰羊卵巢中分离GCs。将试验分为4组:GC(空白对照)、GC+NC(转染siRNA-NC)、GC+si FKBP 5(转染siRNA-FKBP5)、GC+si FKBP 5+LH(促黄体生成素处理转染siRNA-FKBP5)。利用细胞免疫荧光检测FKBP5蛋白在GCs中的定位。通过EdU、Tunel和Western blot检测其对细胞增殖、凋亡的影响;通过Western blot检测其对AKT和ERK信号通路的影响;通过ELISA检测其对E2、P4水平的影响。结果表明,FKBP5表达于GCs细胞质中。FKBP 5表达量随着LH浓度不断升高;5 IU·mL-1 LH作用4 h,FKBP 5表达较高(P<0.001)。干扰FKBP 5后显著抑制了细胞增殖,并显著降低PCNA蛋白表达量(P<0.001);显著促进了细胞凋亡,增加了BAX的表达并减少了Bcl-2的表达(P<0.001);显著抑制了GCs中AKT和ERK信号通路的激活以及E2和P4的分泌(P<0.001)。LH处理部分改善了干扰FKBP 5对GCs的影响(P<0.05)。本研究结果表明,FKBP 5影响绵羊GCs的增殖、凋亡、AKT和ERK信号通路以及E2和P4的分泌,其功能受LH的影响。这些结果为进一步研究FKBP5在绵羊卵泡发育中的作用提供了理论依据。

This study aimed to investigate the potential function of FKBP 5 in ovine ovarian granulosa cells(GCs).GCs were isolated from the ovaries of twenty approximately 2-year-old sexually mature Altay sheep obtained from the Hualing slaughterhouse in Urumqi.The experiment was divided into 4 groups:GC(blank control),GC+NC(transfected with siRNA-NC),GC+si FKBP 5(transfected with siRNA-FKBP 5),and GC+si FKBP 5+LH(treated with luteinizing hormone and transfected with siRNA-FKBP 5).Immunofluorescence was used to detect the localization of FKBP5 protein in GCs.Its effects on cell proliferation and apoptosis were examined using EdU,Tunel assays,and Western Blot analysis.Its impacts on the AKT and ERK signaling pathways were assessed by Western blot,while the levels of E2 and P4 were measured by ELISA.FKBP5 was found to be expressed in the cytoplasm of GCs.Its expression increased with the increasing concentration of LH,reaching a peak at 5 IU·mL-1 LH after 4 h of treatment(P<0.001).Knockdown of FKBP5 significantly inhibited cell proliferation,as evidenced by a reduction in PCNA protein expression(P<0.001).It also significantly promoted cell apoptosis,with an increase in BAX expression and a decrease in Bcl-2 expression(P<0.001).Additionally,FKBP5 knockdown markedly suppressed the activation of the AKT and ERK signaling pathways and the secretion of E2 and P4(P<0.001).LH treatment partially mitigated the effects of FKBP5 knockdown on GCs(P<0.05).The results indicate that FKBP 5 influences the proliferation and apoptosis of ovine GCs,as well as the activation of the AKT and ERK signaling pathways and the secretion of E2 and P4.Its function is modulated by LH.These findings provide a theoretical basis for further research on the role of FKBP5 in ovine follicular development.