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TNF-α水平［（488.30±36.20）μg/L］和IL-1β水平［（62.37±2.50）μg/L］均明显升高（ P<0.001， P=0.018），照射组小鼠血清IL-6水平较对照组小鼠也呈上升趋势，但差异无统计学意义（ P=0.301）。GO_BP分析结果显示，X-RIAEs诱导小鼠卵巢组织表达下调的DEGs主要参与卵泡发育过程，表达上调的DEGs主要参与卵巢组织炎症反应过程，RT-qPCR结果与测序结果一致。IHC结果显示，照射组小鼠卵巢组织SOHLH1阳性表达面积［（23.18±4.00）%］显著低于对照组［（65.90±6.28）%， P=0.005］，而NE阳性表达面积［（30.73±4.00）%］显著高于对照组［（14.47±2.22）%， P=0.024］。 结论:X-RIAEs可诱发卵巢组织炎性反应，并抑制小鼠卵巢卵泡生长及发育过程，进而导致卵巢储备功能下降。

abstractsObjective: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 irradiation group were significantly increased, and the serum level of interleukin (IL)-6 in the irradiation group was also increased compared with the sham group, but the difference was not statistically significant ( P>0.05). The results of GO_BP analysis showed that the down-regulated DGEs were mainly involved in the process of follicular development, and the up-regulated DGEs were involved in the inflammation process. The results of RT-qPCR were consistent with those of sequencing. The immunohistochemistry results showed that the positive expression area of SOHLH1 in the irradiation group [(23.18±4.00)%] was significantly lower than that of the sham group [(65.90±6.28)%, P=0.005], while the positive expression area of NE [(30.73±4.00)%] was significantly higher than that of the sham group [(14.47±2.22)%, P=0.024]. Conclusion:X-RIAEs can induce an inflammatory reaction in ovarian tissue and inhibit the growth and development of ovarian follicles in mice, which leads to a decrease in ovarian reserve.