槲皮素在羊卵巢组织玻璃化冻存中的卵泡保护及抗氧化作用  

Follicle protection and antioxidant effect of quercetin on the cryopreservation of ewe ovarian tissues

作　　者：杨璐恺 蒋利刚[1] 崔妍婷 刘金 韩亦龙[1] 陈超[1] 邓晓惠[1] YANG Lukai;JIANG Ligang;CUI Yanting;LIU Jin;HAN Yilong;CHEN Chao;DENG Xiaohui(Center for Reproductive Medicine,Department of Obstetrics and Gynecology,Qilu Hospital,Cheeloo College of Medicine,Shandong University,Jinan 250012,Shandong,China)

机构地区：[1]山东大学齐鲁医院妇产科不孕不育诊疗中心,山东济南250012

出　　处：《山东大学学报（医学版）》2020年第9期1-7,共7页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金培养基金(ZR2018PC015);山东大学齐鲁医院临床实用新技术发展基金(2017-7)。

摘　　要：目的探讨在冷冻保护剂中添加槲皮素对羊卵巢组织冷冻复苏后卵泡活性的影响。方法将36只性成熟母羊的卵巢皮质组织块随机分配到新鲜对照组(CON组),玻璃化冷冻组(VIT组),添加低、中、高浓度(1、5、10μmol/L)槲皮素的玻璃化冷冻组(VWQ1组、VWQ2组、VWQ3组),将卵巢组织玻璃化冷冻后复苏、培养,测定各组雌激素水平,分析各组的卵泡计数及形态学改变。采用免疫组织化学法进行组织增殖细胞核抗原分析,采用TUNEL法进行组织凋亡分析,采用免疫组织化学法及可见光比色法进行组织抗氧化能力检测。结果 CON组原始卵泡形态正常比例最高、凋亡细胞计数最少(84.1%,P<0.001;13.92±3.88,P<0.001);VWQ1组的原始卵泡正常形态率高于VIT组(76.5%vs 71.7%,P=0.035),凋亡细胞数少于VIT组(50.96±24.28 vs 73.44±35.47,P=0.017),锰超氧化物歧化酶(SOD-2)蛋白表达量、过氧化氢酶(CAT)含量均高于VIT组[0.54(0.53,0.55)vs 0.32(0.29,0.51),P<0.001;5.60±1.49 vs 3.48±1.21,P=0.008];VWQ3组的原始卵泡正常形态率最低(61.2%,P<0.001),凋亡细胞计数最多(127.12±42.46,P<0.001)。结论低浓度槲皮素可以减轻卵巢组织氧化损伤进而保存卵泡活性,但过高浓度的槲皮素对卵泡活性有害。Objective To investigate the effect of quercetin in cryoprotectant on the activity of follicles after cryopreservation of ovarian tissues. Methods Cortical pieces of 36 sexually mature ewe ovaries were randomly assigned to fresh control(CON) group, vitrification(VIT) group, vitrification with low, medium and high concentration(1, 5, 10 μmol/L) of quercetin(VWQ1, VWQ2, VWQ3) groups. The cortical pieces were rewarmed and cultured after vitrification to measure the estrogen levels, count the number of follicles and observe the morphological changes. The proliferating cell nucleus antigen of ovarian tissues was analyzed with immunohistochemistry. Tissue apoptosis was detected with TUNEL, and antioxidant capacity of ovarian tissues was determined with immunohistochemistry and visible light colorimetry. Results The proportion of morphological normal follicles was the highest while the number of apoptotic cells was the lowest in CON group(84.1%, P<0.001;13.92±3.88, P<0.001). Compared with the VIT group, the VWQ1 group had higher rate of morphological normal follicles(76.5% vs 71.7%, P=0.035), fewer number of apoptotic cells(50.96±24.28 vs 73.44±35.47, P=0.017), and higher expressions of manganese superoxide dismutase(SOD-2) and catalase(CAT) [0.54(0.53, 0.55) vs 0.32(0.29, 0.51), P<0.001;5.60±1.49 vs 3.48±1.21, P=0.008]. The VWQ3 group had the lowest rate of morphological normal follicles(61.2%, P<0.001), but the largest number of apoptotic cells(127.12±42.46, P<0.001). Conclusion Low concentration of quercetin in cryoprotection can reduce the oxidative damage on ovarian tissues and thus preserve the follicular activity, but excessively high concentration of quercetin can be harmful.

关 键 词：卵巢组织 冷冻保护剂 抗氧化作用 玻璃化冷冻 羊 

分 类 号：R711.75[医药卫生—妇产科学]