Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro
Cathepsin B (<i>CTSB</i>), a lysosomal cysteine protease’s high expression and activity, has been reported to cause poor-quality embryos in porcine and bovine. Nevertheless, <i>CTSB</i> functions in mice granulosa cells remain to explore. To discuss the <i>CTSB</i>...
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| Autores principales: | , , , , , , |
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| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/775c843a68584cab9d83a37449b12bcf |
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| Sumario: | Cathepsin B (<i>CTSB</i>), a lysosomal cysteine protease’s high expression and activity, has been reported to cause poor-quality embryos in porcine and bovine. Nevertheless, <i>CTSB</i> functions in mice granulosa cells remain to explore. To discuss the <i>CTSB</i> 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 <i>CTSB</i> (siCTSB) for 48 h. Further, mRNA and protein expression of cell proliferation regulators (<i>Myc</i> and <i>cyclin D2</i>), apoptosis regulators (<i>caspase 3</i>, <i>caspase 8</i>, <i>TNF-α</i>, and <i>Bcl2</i>), steroidogenesis-related genes (<i>FSHR</i> and <i>CYP11A1</i>), and autophagy markers (<i>LC3-I</i> and <i>ATG5</i>) were investigated. In addition, the effect of <i>CTSB</i> on steroidogenesis and autophagy was also examined. Flow cytometry analysis assay displayed that silencing of <i>CTSB</i> decreased the early and total apoptosis rate by downregulating <i>TNF-α</i>, <i>caspase 8</i>, and <i>caspase 3</i>, and upregulating <i>Bcl2</i>. By regulating <i>Myc</i> and <i>cyclin D2</i> expression and activating the p-Akt and p-ERK pathways, <i>CTSB</i> 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 <i>LC3-I</i> and <i>ATG5</i> compared to the control group. Herein, we demonstrated that <i>CTSB</i> serves as a proapoptotic agent and plays a critical role in folliculogenesis in female mice by mediating apoptosis, autophagy, proliferation, and steroidogenesis. Hence, <i>CTSB</i> could be a potential prognostic agent for female infertility. |
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