年轻血浆微环境对老年小鼠衰老卵巢的再生作用

doi:10.11958/20250964

天津医药 ›› 2025, Vol. 53 ›› Issue (8): 808-813.doi: 10.11958/20250964

年轻血浆微环境对老年小鼠衰老卵巢的再生作用

刘志鹏¹^,²(), 张小文², 李佩贤², 陈奕豪², 周丹², 杨胜利², 陈柱星², 刘佳¹^,³^,^△()

1 华南理工大学医学院（邮编510006）
2 南方科技大学附属佛山市第一人民医院转化医学研究院
3 大连医科大学基础医学研究院

收稿日期:2025-03-10 修回日期:2025-04-30 出版日期:2025-08-15 发布日期:2025-08-12
通讯作者: ^△E-mail：mcliujia@scut.edu.cn
作者简介:刘志鹏（1999），男，硕士在读，主要从事卵巢衰老方面研究。E-mail：scott_zpliu@163.com
基金资助:
广东省基础与应用基础研究省企联合基金(23202104030001153);佛山市医学研究基金(20230315);佛山市医学研究基金(20240356);佛山市医学研究基金(20240071)

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

LIU Zhipeng¹^,²(), ZHANG Xiaowen², LI Peixian², CHEN Yihao², ZHOU Dan², YANG Shengli², CHEN Zhuxing², LIU Jia¹^,³^,^△()

1 School of Medicine, South China University of Technology, Guangzhou 510006, China
2 the Institution of Tranlational Medicine, the First People’s Hospital of Foshan Affiliated to Southern University of Science and Technology
3 College of Basic Medical Sciences, Dalian Medical University

Received:2025-03-10 Revised:2025-04-30 Published:2025-08-15 Online:2025-08-12
Contact: ^△E-mail：mcliujia@scut.edu.cn

刘志鹏, 张小文, 李佩贤, 陈奕豪, 周丹, 杨胜利, 陈柱星, 刘佳. 年轻血浆微环境对老年小鼠衰老卵巢的再生作用[J]. 天津医药, 2025, 53(8): 808-813.

LIU Zhipeng, ZHANG Xiaowen, LI Peixian, CHEN Yihao, ZHOU Dan, YANG Shengli, CHEN Zhuxing, LIU Jia. The regenerative effect of young plasma microenvironment on aging ovaries of aged mice[J]. Tianjin Medical Journal, 2025, 53(8): 808-813.

摘要/Abstract

摘要：

目的探讨年轻血浆腹腔注射对衰老小鼠生育能力与卵巢功能的影响并分析其潜在的分子机制。方法选取54周龄C57BL/6雌性小鼠与8周龄C57BL/6雄性小鼠，其中雌性小鼠随机均分为生理盐水组、衰老血浆组和年轻血浆组。年轻血浆组与衰老血浆组分别腹腔注射年轻（25~29岁）和年长（45~49岁）女性供体的血浆。每次注射量为500 μL，隔日注射1次，21 d内进行11次注射。生理盐水组小鼠腹腔注射等量生理盐水。末次注射后，予雌雄配种实验评估生育能力；HE染色观察卵巢病理学变化；予超促排卵后获取卵母细胞与受精卵，进行体外培养评估卵母细胞质量及胚胎发育潜能；分析卵巢组织转录组变化，并进行京都基因与基因组百科全书（KEGG）和基因本体（GO）富集分析。结果与生理盐水组和衰老血浆组相比，年轻血浆组小鼠产仔数增多，反映卵母细胞质量的第一极体（PB1）排出率提高，卵母细胞粉碎率降低，卵母细胞的二细胞胚胎转化率及囊胚形成率升高；而衰老血浆组和生理盐水组上述指标无明显差异。转录组学测序发现年轻血浆组小鼠卵巢组织中的差异表达基因主要参与了类固醇生物合成与代谢通路，其中类固醇硫酸酯酶蛋白表达水平显著上调。结论系统性输注年轻血浆可以增强老年小鼠衰老卵巢的生殖潜能，血浆中硫酸化类固醇代谢物可能是恢复卵巢功能和延缓卵巢衰老过程的关键物质。

关键词: 衰老, 卵母细胞, 卵巢功能, 血浆调节, 硫酸化类固醇代谢物, 类固醇生成

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

中图分类号:

R715

图/表 7

图1 血浆注射方案示意图

Fig.1 Schematic diagram of plasma injection protocol

图2 各组卵巢组织切片显微图像（HE染色，×40）

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

图3 各组老年小鼠体内成熟卵母细胞显微图像（×200）

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

表1 各组的卵母细胞数及二细胞和囊胚转化率比较

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^＊＊

图4 各组老年小鼠体内受精卵发育所得二细胞胚胎囊胚（×200）

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

图5 KEGG通路富集分析年轻血浆组的DEGs参与的通路

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

图6 年轻血浆组DEGs的GO功能富集分析结果

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

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年轻血浆微环境对老年小鼠衰老卵巢的再生作用

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

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