Gu Ruihuan, Ge Naidong, Huang Bin, Fu Jing, Zhang Ying, Wang Ningyi, Xu Yan, Li Lu, Peng Xiandong, Zou Yaoyu, Sun Yijuan, Sun Xiaoxi
Department of Shanghai Ji'ai Genetics & IVF Institute, Obstetrics and Gynecology Hospital, Fudan University, Shanghai, China.
Department of Female Fertility Preservation, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China.
Front Genet. 2023 Mar 17;14:1114650. doi: 10.3389/fgene.2023.1114650. eCollection 2023.
This study investigated the effects of a vitrification/warming procedure on the mRNA transcriptome of human ovarian tissues. Human ovarian tissues were collected and processed through vitrification (T-group) and then subjected to RNA sequencing (RNA-seq) analysis, HE, TdT-mediated dUTP nick-end labeling (TUNEL), and real-time quantitative PCR, and the results were compared to those of the fresh group (CK). A total of 12 patients, aged 15-36 years old, with a mean anti-Müllerian hormone level of 4.57 ± 3.31 ng/mL were enrolled in this study. According to the HE and TUNEL results, vitrification effectively preserved human ovarian tissue. A total of 452 significantly dysregulated genes (|log2FoldChange| > 1 and < 0.05) were identified between the CK and T groups. Among these, 329 were upregulated and 123 were downregulated. A total of 372 genes were highly enriched for 43 pathways ( < 0.05), which were mainly related to systemic lupus erythematous, cytokine-cytokine receptor interaction, the TNF signaling pathway, and the MAPK signaling pathway. and were significantly upregulated ( < 0.01), while and were significantly downregulated in the T-group ( < 0.05) compared to the CK group, which agreed with the results of the RNA-seq analysis. These results showed (for the first time to the authors' knowledge) that vitrification can induce changes in mRNA expression in human ovarian tissues. Further molecular studies on human ovarian tissues are required to determine whether altered gene expression could result in any downstream consequences.
本研究调查了玻璃化冷冻/复温程序对人卵巢组织mRNA转录组的影响。收集人卵巢组织并通过玻璃化冷冻处理(T组),然后进行RNA测序(RNA-seq)分析、苏木精-伊红染色(HE)、末端脱氧核苷酸转移酶介导的缺口末端标记法(TUNEL)和实时定量PCR,并将结果与新鲜组(CK)进行比较。本研究共纳入12例年龄在15至36岁之间、平均抗苗勒管激素水平为4.57±3.31 ng/mL的患者。根据HE和TUNEL结果,玻璃化冷冻有效地保存了人卵巢组织。在CK组和T组之间共鉴定出452个显著失调的基因(|log2倍变化|>1且P<0.05)。其中,329个基因上调,123个基因下调。共有372个基因在43条通路中高度富集(P<0.05),这些通路主要与系统性红斑狼疮、细胞因子-细胞因子受体相互作用、TNF信号通路和MAPK信号通路有关。与CK组相比,T组中IL6和CXCL8显著上调(P<0.01),而CCL2和IL1β显著下调(P<0.05),这与RNA-seq分析结果一致。这些结果(据作者所知首次)表明,玻璃化冷冻可诱导人卵巢组织中mRNA表达的变化。需要对人卵巢组织进行进一步的分子研究,以确定基因表达的改变是否会导致任何下游后果。