The regenerative effect of young plasma microenvironment on aging ovaries of aged mice

doi:10.11958/20250964

Abstract

Abstract:

Objective To explore the effect of young plasma intraperitoneal injection on the fertility and ovarian function of aging mice and analyze its potential molecular mechanism. Methods Fifty-four-week-old female C57BL/6 mice and 8-week-old male C57BL/6 mice were selected. Among them, the female mice were randomly divided into three groups: the young plasma group, the aging plasma group and the normal saline group. The young plasma group and the aging plasma group received intraperitoneal injection of plasma from young (25-29 years old) and elderly (45-49 years old) female donors, respectively. Each injection was 500 μL, administered every other day for 2 weeks. The saline group received an equal volume of saline. After the last injection, mating experiments were conducted to evaluate fertility. Ovarian histopathological changes were observed by HE staining. Oocytes and fertilized eggs were collected after superovulation and cultured in vitro to assess oocyte quality and embryo developmental potential. Transcriptomic analysis of ovarian tissue was performed, followed by KEGG and GO enrichment analysis. Results Compared with the normal saline group and the aging plasma group, the number of offspring increased in the young plasma group, which reflected higher extrusion rate of first polar body (PB1), decreased fragmentation rate of oocytes and increased conversion rate of two-cell embryos and increased formation rate of blastocysts. There were no significant differences in these indicators between the aging plasma group and the normal saline group. Transcriptomic sequencing revealed that the differentially expressed genes in ovarian tissue of the young plasma group were mainly involved in steroid biosynthesis and metabolic pathways. Among them, the expression level of steroid sulfatase protein was significantly upregulated. Conclusion Systemic infusion of young plasma enhances the reproductive potential of aging ovaries in elderly mice. The sulfated steroid metabolites in plasma may be key substances in restoring ovarian function and delaying the process of ovarian aging.

Key words: aging, oocytes, ovarian function, plasma regulation, sulfated steroid metabolites, steroidogenesis

CLC Number:

R715

Figures/Tables 7

Fig.1 Schematic diagram of plasma injection protocol

Fig.2 Microscopic images of ovarian tissue sections in each group (HE staining, ×40)

Fig.3 Micrographs of in vivo matured oocytes from aged mice in each experimental group (×200)

Tab.1 Comparison of the number of oocytes and the conversion rates of two-cell and blastocysts between three groups of aged mice （n=6，$\bar{x}±s$）

组别	卵母细胞总数/个	PB1排出率/%	细胞粉碎率/%	二细胞胚胎转化率/%	囊胚形成率/%
生理盐水组	12.1±4.2	29.6±10.4	49.2±2.8	39.3±3.5	30.1±6.7
衰老血浆组	7.0±3.4^a	20.2±19.6	67.3±19.8	27.1±13.0	20.4±12.6
年轻血浆组	15.5±4.0^b	42.3±12.2^ab	35.4±14.7^ab	52.5±4.3^ab	42.7±11.8^ab
F	6.602^＊	4.443^＊	7.298^＊＊	6.233^＊	12.014^＊＊

Fig. 4 In vivo fertilized embryos from aged mice across experimental groups: two-cell embryos and blastocysts (×200)

Fig.5 KEGG pathway enrichment analysis of the pathways involved in DEGs in the young plasma group

Fig.6 Results of GO functional enrichment analysis of DEGs in the young plasma group

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