Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Hubei Province's Engineering Research Center in Buffalo Breeding and Products, Wuhan 430070, China.
Int J Mol Sci. 2021 Oct 31;22(21):11827. doi: 10.3390/ijms222111827.
Cathepsin B (), a lysosomal cysteine protease's high expression and activity, has been reported to cause poor-quality embryos in porcine and bovine. Nevertheless, functions in mice granulosa cells remain to explore. To discuss the functional role in follicular dynamics, we studied apoptosis, proliferation, cell cycle progression, and related signaling pathways in primary mouse granulosa cells transfected with small interference RNA specific to (siCTSB) for 48 h. Further, mRNA and protein expression of cell proliferation regulators ( and ), apoptosis regulators (, , , and ), steroidogenesis-related genes ( and ), and autophagy markers ( and ) were investigated. In addition, the effect of on steroidogenesis and autophagy was also examined. Flow cytometry analysis assay displayed that silencing of decreased the early and total apoptosis rate by downregulating , , and , and upregulating . By regulating and expression and activating the p-Akt and p-ERK pathways, knockdown increased GC proliferation and number. A significant decline in estradiol and progesterone concentrations was observed parallel to a significant decrease in autophagy-related markers and compared to the control group. Herein, we demonstrated that serves as a proapoptotic agent and plays a critical role in folliculogenesis in female mice by mediating apoptosis, autophagy, proliferation, and steroidogenesis. Hence, could be a potential prognostic agent for female infertility.
组织蛋白酶 B ()，一种溶酶体半胱氨酸蛋白酶，其高表达和活性已被报道会导致猪和牛的胚胎质量差。然而，其在小鼠颗粒细胞中的功能仍有待探索。为了探讨其在卵泡动态中的功能作用，我们研究了转染针对组织蛋白酶 B 的小干扰 RNA (siCTSB) 48 小时后，原代小鼠颗粒细胞中的细胞凋亡、增殖、细胞周期进程和相关信号通路。此外，还研究了细胞增殖调节剂 ()、凋亡调节剂 ()、()、()、()、()和 ()、类固醇生成相关基因 ()和自噬标记物 ()的 mRNA 和蛋白表达。此外，还研究了对类固醇生成和自噬的影响。流式细胞术分析显示，通过下调和，上调和，沉默会降低早期和总凋亡率。通过调节和的表达并激活 p-Akt 和 p-ERK 通路，CTSB 敲低会增加 GC 的增殖和数量。与对照组相比，雌二醇和孕酮浓度显著下降，同时自噬相关标记物和显著下降。因此，我们证明组织蛋白酶 B 通过介导细胞凋亡、自噬、增殖和类固醇生成，作为一种促凋亡剂在雌性小鼠的卵泡发生中发挥关键作用。因此，组织蛋白酶 B 可能是女性不孕的一个潜在预后标志物。
