基于Nrf2/HO-1信号通路研究白藜芦醇在小鼠卵巢冷冻复苏中的保护作用及机制  

Protective Effects and Mechanisms of Resveratrol in Mouse OvarianCryopreservation Based on Nrf2/HO-1 Signaling Pathway

作　　者：张珦 李惠 章宜芬 郑燕影 孙怡 陈劼 储乐乐 应葳 李长印 郑艳莉 ZHANG Xiang;LI Hui;ZHANG Yi-Fen;ZHEN Yan-Ying;SUN Yi;CHEN Jie;CHU Le-Le;YING Wei;LI Chang-Yin;ZHENG Yan-Li(Dept.of Pathology,The Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210000 Jiangsu,China)

机构地区：[1]南京中医药大学附属医院病理科,江苏南京210000

出　　处：《广州中医药大学学报》2023年第5期1221-1227,共7页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：江苏省妇幼健康科研项目(编号:F201836)。

摘　　要：【目的】探讨白藜芦醇在新生小鼠卵巢冷冻复苏中的保护作用及其作用机制。【方法】行冷冻复苏操作,在冷冻复苏前使用白藜芦醇预处理新生小鼠卵巢,在冷冻复苏液中加入白藜芦醇。末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测卵巢组织中细胞凋亡情况,免疫组织化学法观察卵巢组织中Cleaved-Caspase-3表达水平,采用苏木素-伊红(HE)染色法观察移植卵巢卵泡数量及病理情况,蛋白免疫印迹(Western Blot)法检测核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)、多聚ADP核糖多聚糖(PARP)、剪切型PARP(Cleaved-PARP)蛋白表达水平。【结果】白藜芦醇组冷冻复苏后卵巢组织中细胞凋亡减少,移植卵巢卵泡数目显著升高,卵巢组织中Cleaved-Caspase-3表达水平显著降低,抗氧化蛋白Nrf2蛋白表达水平明显升高,凋亡蛋白Cleaved-PARP/PARP比值明显降低,与模型对照组比较,差异均有统计学意义(P<0.05),抗氧化蛋白HO-1表达水平升高,但与模型对照组比较,差异无统计学意义(P>0.05)。【结论】白藜芦醇在小鼠卵巢冷冻复苏移植过程中可减轻卵巢氧化应激、抑制卵巢细胞凋亡,对卵巢起保护作用。Objective To investigate the protective effect of resveratrol in neonatal mouse ovarian cryopreservation and its mechanism of action.Methods The neonatal mouse ovaries were pretreated with resveratrol prior to freezing,and resveratrol was added to the freezing and resuscitation solution.Apoptosis in ovarian tissue was detected by dUTP notch end labeling(TUNEL)mediated by terminal deoxynucleotide transferase,the expression level of Cleaved-Caspase-3 in ovarian tissue was measured by immunohistochemistry,the number of follicles and pathology in transplanted ovaries were observed by hematoxylin-eosin(HE)staining,and the expression levels of nuclear factor E2-related factor 2(Nrf2),heme oxygenase(HO-1),poly-ADP-ribose polysaccharide(PARP)and Cleaved-PARP were measured by Western Blot.Results After ovarian tissue cryopreservation,cell apoptosis was decreased and the number of follicles in transplanted ovaries was significantly increased in the resveratrol group,Cleaved Caspase-3 expression level was significantly decreased,antioxidant protein Nrf2 protein expression level was significantly increased,apoptosis protein Cleaved PARP/PARP ratio was significantly decreased,the differences being statistically significant as compared with the model control group(P<0.05),but antioxidant protein HO-1 expression level was enhanced,the difference being statistically insignificant as compared with the model control group(P>0.05).Conclusion Resveratrol protects the ovary by reducing oxidative stress and inhibiting ovarian apoptosis during cryo-resuscitation transplantation in mice.

关 键 词：白藜芦醇 卵巢 冷冻复苏 异体移植 生育力 Nrf2/HO-1信号通路 氧化应激 细胞凋亡 小鼠 

分 类 号：R285.5[医药卫生—中药学]