OPN5对鸭颗粒细胞凋亡、增殖及类固醇激素生成的影响

doi:10.11843/j.issn.0366-6964.2021.012.015

Abstract

Abstract: This study aimed to characterize the effects of neuropsin (OPN5) on apoptosis, prolife-ration and steroid hormone production of granulosa cells of duck. The duck granulosa cells were isolated and transfected with OPN5 overexpression plasmid and OPN5 siRNA, respectively. After continuous culture for 72 h (n=6), both proliferation and apoptosis of granulosa cell were detected using EdU cell proliferation assay, flow cytometry, Annexin V-FITC. The mRNA and protein expression levels, as well as hormone secretion were investigated by RT-PCR, Western blot and ELISA, respectively. The results showed that overexpression of OPN5 promoted prolife-ration and inhibited apoptosis of follicular granulosa cells in ducks. Moreover, overexpression of OPN5 significantly promoted the expression of GnRHR, FSHR, LHR, inhibited the expression of GnIH and GnIHR (P<0.01), significantly or highly significantly upregulated the mRNA levels of StAR, CYP11A1, 3β-HSD, CYP17A1 and CYP19A1 (P<0.05 or P<0.01), significantly increased the protein expression levels of OPN5, 3β-HSD and CYP19A1 (P<0.05), highly significantly increased the secretion levels of E2 and P4 (P<0.01). However, the levels of INHβ was highly significantly decreased (P<0.01). OPN5 knockdown with siRNA could effectively downregulate OPN5 expression in transfected cells (P<0.01), inhibit proliferation and promote apoptosis of follicular granulosa cells. siRNA transfection significantly downregulated the expression levels of GnRH, GnRHR, FSHR, LHR (P<0.01), promoted the expression of GnIH and GnIHR (P<0.01), significantly inhibited the expression of StAR, CYP11A1, CYP17A1 (P<0.01). OPN5 knockdown significantly decreased the protein expression levels of OPN5, 3β-HSD and CYP19A1 (P<0.05), highly significantly decreased the secretion levels of E2 and P4 (P<0.01), and highly significantly increased the level of INHβ (P<0.01). In conclusion, OPN5 promotes the proliferation and inhibits the apoptosis of duck follicular granulosa cells, and promotes the production and secretion of steroid hormones in granulosa cells.

Key words: OPN5, duck, granulosa cell, apoptosis, proliferation, steroid hormone

CLC Number:

S834.83

LIUFU Sui, PAN Jianqiu, JIANG Danli, SHEN Xu, XU Danning, TIAN Yunbo, HUANG Yunmao. Effects of OPN5 on Apoptosis, Proliferation and Steroid Hormone Synthesis of Duck Granulosa Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3480-3491.

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Effects of OPN5 on Apoptosis, Proliferation and Steroid Hormone Synthesis of Duck Granulosa Cells

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