低强度超声缓解顺铂所致小鼠卵巢损伤的实验研究  

Alleviation of cisplatin-induced ovarian damage in mice by low-intensity ultrasound:an experimental study

作　　者：周圆圆[1] 周怡[1] 段亚阳 张怡卿 朱峰宇 张超学[1] Zhou Yuanyuan;Zhou Yi;Duan Yayang;Zhang Yiqing;Zhu Fengyu;Zhang Chaoxue(The First Affiliated Hospital of Anhui Medical University,Key Laboratory of Gametes and Reproductive Tract Anomalies Research,National Health and Wellness Commission,Hefei 230022,China)

机构地区：[1]安徽医科大学第一附属医院、国家卫生健康委配子及生殖道异常研究重点实验室,合肥230022

出　　处：《中华医学超声杂志(电子版)》2024年第12期1132-1141,共10页Chinese Journal of Medical Ultrasound(Electronic Edition)

摘　　要：目的探讨低强度脉冲超声(LIPUS)对顺铂所致小鼠卵巢细胞损伤的缓解作用。方法将40只ICR小鼠分为对照组(n=12)、顺铂组(n=14)、顺铂+LIPUS组(n=14),对照组小鼠腹腔注射生理盐水,顺铂组小鼠腹腔注射顺铂,顺铂+LIPUS组小鼠腹腔顺铂注射、次日LIPUS照射,周期共2周。取小鼠卵巢行HE染色、RNA测序、免疫组化、TUNEL染色、蛋白质免疫印迹(western blot)及实时荧光定量聚合酶链反应(qPCR),进行卵泡计数,检测卵巢中凋亡、铁死亡及氧化应激等细胞生理事件相应细胞因子表达水平。结果与对照组相比,顺铂组卵巢体积缩小;与顺铂组相比,顺铂+LIPUS组小鼠的卵巢体积增大。对照组、顺铂组与顺铂+LIPUS组的卵巢重量比较[(8.8±1.34)mg vs(2.8±0.45)mg vs(5.6±1.35)mg],差异有统计学意义(P<0.05)。与对照组相比,顺铂组原始卵泡、初级卵泡、次级卵泡及有腔卵泡的数目减少;与顺铂组相比,顺铂+LIPUS组相应各级卵泡数目增多(P<0.05)。LIPUS治疗与否差异基因主要富集在细胞生理水平事件上,如细胞凋亡、铁死亡及氧化应激方面。与对照组相比,顺铂组Bax、p-H2AX、TUNEL信号、3-Nit、4-HNE及8-OHdG表达增加,Bcl2、SIRT1、SIRT2、Nrf2、FPN、GPX4、Fth、Ftl等因子表达减少(P<0.05);与顺铂组相比,顺铂+LIPUS组Bax、p-H2AX、TUNEL信号、3-Nit、4-HNE及8-OHdG表达减少,Bcl2及铁死亡相应各因子除SIRT2外表达均增加(P<0.05)。结论LIPUS可以缓解顺铂所致小鼠卵巢损伤,其机制可能是通过调控凋亡、铁死亡及氧化应激细胞因子的表达。Objective To investigate the repairing effect of low-intensity ultrasound(LIPUS)on cisplatin-induced ovarian cell damage in mice.Methods Forty ICR mice were divided into a control group(n=12),a cisplatin group(n=14),and a cisplatin+LIPUS group(n=14).Control mice were injected intraperitoneally with saline,mice in the cisplatin group were injected intraperitoneally with cisplatin,and the cisplatin+LIPUS group underwent intraperitoneal cisplatin injection,followed by LIPUS irradiation on the next day.The treatment cycle was 2 weeks long.Mouse ovary tissues were taken for HE staining for follicle counting.RNA sequencing,immunohistochemistry,TUNEL staining,protein immunoblotting(Western blot),and real-time fluorescence quantitative polymerase chain reaction(qPCR)were performed to detect the expression of cytokines in different cellular physiological events in the ovaries such as apoptosis,iron death,and oxidative stress.Results Compared to the control group,ovarian size was reduced in the cisplatin group;compared to the cisplatin group,ovary size was increased in the cisplatin+LIPUS group.Comparison of ovarian weights in the control,cisplatin,and cisplatin+LIPUS groups(8.8±1.34 vs 2.8±0.45 vs 5.6±1.35)revealed statistically significant differences(P<0.05).Compared to the control group,the number of primordial follicles,primary follicles,secondary follicles and antral follicles was reduced in the cisplatin group;compared with the cisplatin group,the number of follicles at the corresponding levels was increased in the cisplatin+LIPUS group(P<0.05).Between mice with LIPUS treatment and controls,differential genes were mainly enriched in events at the cellular physiological level,such as apoptosis,iron death,and oxidative stress.Compared to the control group,Bax,p-H2AX,TUNEL signal,3-Nit,4-HNE,and 8-OHdG levels were increased,while the levels of factors such as Bcl2,SIRT1,SIRT2,Nrf2,FPN,GPX4,Fth,and Ftl were reduced in the cisplatin group(P<0.05);compared with the cisplatin group,the levels of Bax,p-H2AX,TUNEL signal,

关 键 词：低强度超声 卵巢 顺铂 凋亡 铁死亡 氧化应激 小鼠 

分 类 号：R445.1[医药卫生—影像医学与核医学] R711.75[医药卫生—诊断学]