Effects and Mechanisms of Prdx4 on Apoptosis of Granulosa Cell in Mice

doi:10.12280/gjszjk.20210596

Abstract

Abstract:

Objective: To explore the effects and mechanisms of peroxiredoxin 4 (Prdx4) on the apoptosis of granulosa cells in mice. Methods： The localization of Prdx4 in the human ovarian granulosa cellline (KGN) was detected by immunofluorescence staining. The expression of Prdx4 in granulosa cell was down-regulated by a Prdx4-siRNA, which was evaluated by Western blotting (WB). Apoptosis-related markers including B cell lymphoma 2 (Bcl-2), Bcl-2 associated X (Bax) and cleaved caspase 3 were also tested by WB. After that, the endoplasmic reticulum stress (ER stress)-related proteins including binding immunoglobulin protein (BIP), activating transcription factor 4 (ATF4), ATF6 and C/EBP homologous protein (CHOP) were then detected. Apoptosis was examined by flow cytometry. Electron microscopy was used to characterize the morphological changes of endoplasmic reticulum (ER). Results： In KGN cells, Prdx4 was mainly localized in ER. The expression of Prdx4 protein in the Prdx4-siRNA group was significantly reduced (P<0.01). Flow cytometry analysis revealed that the apoptosis ratio was increased significantly in the Prdx4-siRNA group (P<0.05). Compared with the control group, the Prdx4-siRNA group had the increased expressions of the proapoptotic proteins, Bax and cleaved caspase 3 (P<0.01) and the decreased expression of antiapoptotic protein, Bcl-2 (P<0.05). The electron microscopy showed that ER was swelled and dilated significantly in the Prdx4-siRNA group. Interestingly, the Prdx4-siRNA group was showed the significant increases in the expressions of BIP, ATF4 and CHOP protein (P<0.05). Conclusions： Knockdown expression of Prdx4 in granulosa cells would induce the cell apoptosis by the enhanced ER stress.

Key words: Peroxiredoxin Ⅲ, Endoplasmic reticulum stress, Ovary, Granulosa cells, Apoptosis

ZOU Xiao-fei, LIANG Xiu-ru, YAN Zheng-jie, CUI Yu-gui, LIU Jia-yin, MENG Yan. Effects and Mechanisms of Prdx4 on Apoptosis of Granulosa Cell in Mice[J]. Journal of International Reproductive Health/Family Planning, 2022, 41(2): 89-93.

Figures/Tables 6

References 23

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组别	n	cleaved caspase 3	Bax	Bcl-2
空白对照组	3	0.590±0.081	0.866±0.027	1.025±0.051
Prdx4-siRNA处理组	3	1.282±0.154	1.204±0.049	0.787±0.110
t		6.909	10.450	3.409
P		0.002	0.001	0.027

组别	n	cleaved caspase 3	Bax	Bcl-2
空白对照组	3	0.590±0.081	0.866±0.027	1.025±0.051
Prdx4-siRNA处理组	3	1.282±0.154	1.204±0.049	0.787±0.110
t		6.909	10.450	3.409
P		0.002	0.001	0.027

组别	n	Prdx4	BIP	ATF6	ATF4	CHOP
空白对照组	3	1.057±0.126	0.563±0.169	0.943±0.106	0.373±0.139	0.567±0.145
Prdx4-siRNA处理组	3	0.469±0.080	0.967±0.162	0.671±0.486	0.959±0.149	1.013±0.162
t		6.859	2.982	0.948	4.975	3.549
P		0.002	0.041	0.397	0.008	0.024

组别	n	Prdx4	BIP	ATF6	ATF4	CHOP
空白对照组	3	1.057±0.126	0.563±0.169	0.943±0.106	0.373±0.139	0.567±0.145
Prdx4-siRNA处理组	3	0.469±0.080	0.967±0.162	0.671±0.486	0.959±0.149	1.013±0.162
t		6.859	2.982	0.948	4.975	3.549
P		0.002	0.041	0.397	0.008	0.024