Experimental Study on the Involvement of Lipid Peroxidation and Ferroptosis in the Regulation of Senescent Ovarian Granulosa Cells

doi:10.12280/gjszjk.20260180

Abstract

Abstract:

Objective: To compare the levels of lipid peroxidation, iron accumulation and ferroptosis sensitivity between normal and senescent ovarian granulosa cells by the in vivo and in vitro experiments. Methods: The mitochondrial status and iron accumulation in the ovarian tissues of female C57BL/6 mice aged 3 months (reproductive active period) and 12 months (reproductive senescence) were detected in the in vivo experiment. In the in vitro experiment, the SVOG cell, the human ovarian granulosa cell line, was used to develope the senescence model by thapsigargin (TG). The cellular senescence-related markers, lipid peroxidation and ferroptosis-related markers were detected. Senescence-associated proteins such as P16, P21 and TP53 were included. Pro-inflammatory cytokines of the senescence-associated secretory phenotype (SASP), including interleukin-1A (IL-1A), IL-1B, IL-6 and C-X-C chemokine ligand 8 (CXCL8) were analyzed. The ferroptosis and lipid peroxidation indicators were also tested, including the expression levels of ferroptosis-related proteins [glutathione peroxidase 4 (GPX4), transferrin receptor (TFRC), and nuclear receptor coactivator 4 (NCOA4)], and the levels of reactive oxygen species (ROS), malondialdehyde (MDA), and ferrous ion (Fe²⁺). Results: The in vivo experiment confirmed a significant increase in the iron accumulation in senescence ovaries (P<0.01). The in vitro experiment confirmed that TG successfully induced the senescence model of SVOG cells and reduced cell viability (P<0.05). In the senescent cells, the expression of the ferroptosis-related protein GPX4 was decreased (P<0.000 1), while the expressions of TFRC and NCOA4 were increased (P<0.01). Furthermore, the levels of ROS, MDA and Fe²⁺, were all increased (P<0.01). Conclusions: Senescent ovarian granulosa cells display the iron accumulation and the elevated lipid peroxidation, and the increased ferroptosis sensitivity, indicating that ferroptosis participates in the regulation of ovarian granulosa cell aging.

Key words: Ovary, Cellular senescence, Ferroptosis, Lipid peroxidation, Granulosa cells

DONG Si-rui, MENG Yan. Experimental Study on the Involvement of Lipid Peroxidation and Ferroptosis in the Regulation of Senescent Ovarian Granulosa Cells[J]. Journal of International Reproductive Health/Family Planning, 2026, 45(3): 177-183.

Figures/Tables 5

References 22

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基因名称	序列
IL-1A	上游：5'-AATCATCAAGCCTAGGTCAGC-3'
	下游：5'-CTTCATCTTGGGCAGTCACA-3'
IL-1B	上游：5'-GTTGAAAGATGATAAGCCCACT-3'
	下游：5'-GTTATATCCTGGCCGCCTT-3'
IL-6	上游：5'-CACACAGACAGCCACTCACC-3'
	下游：5'-ATTTTCACCAGGCAAGTCTCC-3'
CXCL8	上游：5'-CAGTTTTGCCAAGGAGTGCTA-3'
	下游：5'-TTTTCCTTGGGGTCCAGACA-3'
GAPDH	上游：5'-GGAGCGAGATCCCTCCAAAAT-3'
	下游：5'-GGCTGTTGTCATACTTCTCATGG-3'

基因名称	序列
IL-1A	上游：5'-AATCATCAAGCCTAGGTCAGC-3'
	下游：5'-CTTCATCTTGGGCAGTCACA-3'
IL-1B	上游：5'-GTTGAAAGATGATAAGCCCACT-3'
	下游：5'-GTTATATCCTGGCCGCCTT-3'
IL-6	上游：5'-CACACAGACAGCCACTCACC-3'
	下游：5'-ATTTTCACCAGGCAAGTCTCC-3'
CXCL8	上游：5'-CAGTTTTGCCAAGGAGTGCTA-3'
	下游：5'-TTTTCCTTGGGGTCCAGACA-3'
GAPDH	上游：5'-GGAGCGAGATCCCTCCAAAAT-3'
	下游：5'-GGCTGTTGTCATACTTCTCATGG-3'