小鼠动情周期卵巢、子宫及输卵管中氧化应激水平及意义

The Role and Significance of ROS in the Ovary,Uterus and Oviduct in Mice during Estrous Cycle

目的探讨小鼠动情周期不同阶段氧化应激(OS)的变化规律及对雌性生殖的作用。方法应用ELISA方法检测动情周期各阶段血清中的丙二醛(MDA)、还原型谷胱甘肽(GSH)、8-羟基脱氧鸟苷(8-OHdG)及超氧化物歧化酶(SOD),应用免疫组化方法检测DNA修复蛋白8-羟基鸟嘌呤DNA糖苷酶1(OGG1)及超氧化物歧化酶1(SOD1)在动情周期各阶段输卵管和子宫角中的表达定位,实时荧光定量(PCR)及Western blot检测动情周期各阶段卵巢和子宫角中的OGG1及SOD1基因及蛋白表达水平。结果ELISA结果显示,动情期血清MDA和8-OHdG高于间情期、动情后期及动情前期(P均<0.05),动情前期血清GSH高于动情期、动情后期及间情期(P均<0.05),但是血清总SOD水平在动情周期四个阶段间差异无统计学意义(P均>0.05);PCR和Western blot结果显示,动情期卵巢和子宫角中的SOD1和OGG1 mRNA和蛋白水平低于间情期(P均<0.05),而免疫组化结果显示:OGG1和SOD1蛋白主要定位于子宫内膜腔上皮和腺上皮细胞,定位于输卵管上皮细胞,且SOD1和OGG1在动情期的输卵管中高表达。结论动情周期中OS水平的动态变化提示OS可能参与雌性生殖,特别是排卵、受精过程,且与动情周期雌孕激素的变化密切相关。

Objective The present work aimed to investigate the pattern and function of OS in mouse ovary,uterus and oviduct during the estrous cycle.Methods The serum,ovaries and uteri from proestrus,estrous,metestrus and diestrus were collected and serum malondialdehyde(MDA),glutathione(GSH),8-hydroxy-desoxyguanosine(8-OHdG)and super oxidase dismutase(SOD)were detected by ELISA,and the expression of 8-oxoguanine DNA glycosylase(OGG1)and SOD1 were checked by real-time PCR and western blot.Results The results of ELISA suggested that the serum MDA and 8-OHdG in the estrous phage was significantly higher than in others(P<0.05),the serum GSH in the proestrus was significantly higher than in others(P<0.05),but the serum total SOD levels was no significantly difference in the four stages during estrous cycle(P>0.05).The PCR and Western blot results suggested that the expression of OGG1,SOD1 mRNA and protein in the ovary during estrous phage was significantly lower than diestrus phage(P<0.05).The immunohistochemical results suggested that the OGG1 and SOD1 mainly localized in the endometrial and glandular epithelial cell,localized in the oviduct epithelial cell,the expression of OGG1 and SOD1 protein remarkably higher than in others from immunohistochemical observation.Conclusion Taken together,the change of oxidative stress during estrous cycle might be involved in the female reproduction,such as ovulation,fertilization,and the OS might be closely related with the change of estrogen and progesterone during estrous cycle.