College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.
Theriogenology. 2025 Jan 1;231:62-72. doi: 10.1016/j.theriogenology.2024.10.010. Epub 2024 Oct 13.
As a member of peroxisome proliferator-activated receptor (PPAR) family, PPARG has been reported to be involved in glucolipid metabolism in various species. However, the function of PPARG in estradiol (E2) synthesis, apoptosis, and proliferation in goat ovarian granulosa cells (GCs) is unclear. In this study, we found that goat PPARG was expressed in GCs of all grades of follicles, and localized in the cytoplasm and nucleus of GCs. Transfection of small interfering RNA-PPARG2 (si-PPARG2) decreased E2 synthesis and the abundances of HSD3B and CYP19A1 mRNA and protein. It also promoted cell apoptosis with significant increases in the ratio of BAX/BCL-2 and Caspase3 mRNA and protein. Meanwhile, cell cycle was inhibited by si-PPARG2 transfection, accompanied by decreased mRNA levels of CDK4, CKD6, MYC, CCND1, CCND2, PCNA, and CCNB, increased mRNA level of P53, and decreased protein levels of CDK4, MYC, and CCND1. Furthermore, PPARG interference affected the mRNA and protein abundances of CREB as well as the phosphorylation of CREB but not PKA. In conclusion, our data suggest that PPARG plays an important role in regulating E2 synthesis, cell apoptosis, and proliferation of goat GCs, including the CREB expression and phosphorylation. These results provide evidences for the in-depth study of PPARG in the regulation of goat GCs function.
作为过氧化物酶体增殖物激活受体 (PPAR) 家族的一员,PPARG 已被报道参与多种物种的糖脂代谢。然而,PPARG 在山羊卵巢颗粒细胞 (GC) 中的雌二醇 (E2) 合成、凋亡和增殖中的功能尚不清楚。在本研究中,我们发现山羊 PPARG 在各级卵泡的 GC 中表达,并定位于 GC 的细胞质和细胞核中。PPARG2 小干扰 RNA 转染 (si-PPARG2) 降低了 E2 合成以及 HSD3B 和 CYP19A1 mRNA 和蛋白的丰度。它还促进了细胞凋亡,BAX/BCL-2 和 Caspase3 mRNA 和蛋白的比值显著增加。同时,si-PPARG2 转染抑制了细胞周期,伴随 CDK4、CCKD6、MYC、CCND1、CCND2、PCNA 和 CCNB 的 mRNA 水平降低,P53 的 mRNA 水平升高,以及 CDK4、MYC 和 CCND1 的蛋白水平降低。此外,PPARG 干扰影响了 CREB 的 mRNA 和蛋白丰度以及 CREB 的磷酸化,但不影响 PKA。总之,我们的数据表明,PPARG 在调节山羊 GCs 的 E2 合成、细胞凋亡和增殖中发挥重要作用,包括 CREB 的表达和磷酸化。这些结果为深入研究 PPARG 在调节山羊 GCs 功能中的作用提供了证据。