X射线辐射远端效应诱发小鼠卵巢储备功能下降的实验研究  

Diminished ovarian reserve induced by X-ray radiation-induced abscopal effects in mice:an experimental study

作　　者：周莹 郭妍妍[2] 李岩 叶舒婷 张步美[1] 黎小佩 王建梅[1] 杜湧瑞 Zhou Ying;Guo Yanyan;Li Yan;Ye Shuting;Zhang Bumei;Li Xiaopei;Wang Jianmei;Du Yongrui(Department of Family Planning&Center for Reproductive Medicine,the Second Hospital of Tianjin Medical University,Tianjin 300211,China;Department of Radiation Oncology,the Second Hospital of Tianjin Medical University,Tianjin 300211,China)

机构地区：[1]天津医科大学第二医院计划生育科&辅助生殖中心,天津300211 [2]天津医科大学第二医院放射治疗科,天津300211

出　　处：《中华生殖与避孕杂志》2023年第7期703-712,共10页Chinese Journal of Reproduction and Contraception

基　　金：天津医科大学第二医院青年科研基金(2020ydey02)。

摘　　要：目的探讨X射线辐射远端效应(X-ray radiation-induced abscopal effects,X-RIAEs)对小鼠卵巢储备的影响及可能的作用机制。方法16只动情周期规律的6~8周龄雌性C57BL/6J小鼠分为对照组和照射组,8只/组,照射组小鼠麻醉后每日给予胸部局部区域8 Gy X射线照射,连续照射3 d,对照组小鼠仅给予麻醉处理。照射结束21 d后,检测两组小鼠动情周期、血清激素及促炎性因子水平、卵巢组织形态学变化;利用转录组测序技术(ribonucleic acid sequencing,RNA-seq)检测小鼠卵巢组织RNA转录组表达情况,筛选差异表达基因(differentially expressed genes,DEGs)后行基因本体论-生物学过程(gene ontology-biological processes,GO_BP)分析,通过应用实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)验证测序结果,免疫组织化学染色(immunohistochemistry,IHC)检测精卵发生特异性碱性螺旋环螺旋蛋白1(spermatogenesis-and oogenesis-specific basic helix-loop-helix-containing protein 1,SOHLH1)和中性粒细胞弹性蛋白酶(neutrophil elastase,NE)在卵巢组织内表达和定位情况。结果照射组小鼠动情周期紊乱,主要停滞于间期,照射组小鼠始基卵泡数量[10.50(1.25,12.75)]及生长卵泡数量[4.50(2.50,9.00)]均显著少于对照组小鼠[60.00(30.00,90.25),P<0.001;18.50(18.00,20.75),P<0.001],差异均具有统计学意义,而闭锁卵泡数量[56.00(45.25,98.75)]明显多于对照组[12.50(5.25,20.25)],差异具有统计学意义(P<0.001);照射组小鼠血清雌二醇水平[(70.28±5.27)pmol/L]、抗苗勒管激素水平[(104.00±6.98)μg/L]均明显低于对照组小鼠[(97.58±7.25)pmol/L,P=0.016;(129.70±8.39)μg/L,P=0.046],而照射组小鼠卵泡刺激素水平与对照组小鼠相比,差异无统计学意义(P=0.996);与对照组小鼠血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平[(31.61±12.89)μg/L]、白细胞介素(interleukin,IL)-1β水平[(52.75±2.06)μg/L]相比,照射组小鼠血清TNObjective To investigate the effect and possible mechanism of the X-ray radiation-induced abscopal effects(X-RIAEs)on the ovarian reserve of mice.Methods Totally sixteen female C57BL/6J mice aged 6-8 weeks with regular estrous cycle were randomly divided into the sham group(n=8)and irradiation group(n=8).After anesthesia,the mice in the irradiation group were irradiated with 8 Gy X-ray on the local area of the chest every day for 3 d,while the mice in the sham group were not irradiated.After irradiation 21 d,the estrous cycle,serum hormones,serum pro-inflammatory factors,and ovarian morphological changes were detected.Ribonucleic acid sequencing(RNA-seq)was used to detect the expression of transcriptional levels in mouse ovarian tissues.The differentially expressed genes(DEGs)were screened and analyzed by gene ontology-biological process(GO_BP).Real-time quantitative polymerase chain reaction(RT-qPCR)verified the sequencing results.The expression and localization of spermatogenesis-and oogenesis-specific basic helix-loop-helix-containing protein 1(SOHLH1)and neutrophil elastase(NE)in ovarian tissues were detected by immunohistochemistry(IHC).Results Compared with mice in the sham group,the irradiation group had a disordered estrous cycle,reduced primordial follicles[10.50(1.25,12.75)vs.60.00(30.00,90.25),P<0.001]and growing follicles[(4.50(2.50,9.00)vs.18.50(18.00,20.75),P<0.001],significantly increased atretic follicles[56.00(45.25,98.75)vs.12.50(5.25,20.25),P<0.001].The levels of serum estradiol[(70.28±5.27)pmol/L]and anti-Müllerian hormone[(104.00±6.98)μg/L]in the irradiation group were significantly lower than those in the sham group[(97.58±7.25)pmol/L,P=0.016;(129.70±8.39)μg/L,P=0.046],but the follicle-stimulating hormone(FSH)level in the irradiation group was not significantly different from that in the sham group(P>0.05).Compared with the sham group,the serum levels of TNF-α[(488.30±36.20)μg/L vs.(31.61±12.89)μg/L,P<0.001]and IL-1β[(62.37±2.50)μg/L vs.(52.75±2.06)μg/L,P=0.018]in the irradi

关 键 词：放射治疗 卵巢储备 生育力 远端效应 始基卵泡 

分 类 号：R818[医药卫生—放射医学]