卢晓声 吴昊 习海涛 黄朝霞 吕杰强

[摘要] 目的 探討蛇床子素（OST）对卵巢氧化损伤的保护作用。 方法 体外培养小鼠卵巢组织，用过氧化氢（H2O2）构建卵巢组织氧化损伤模型（模型组）;药物组以20 μg/mL、40 μg/mL和80 μg/mL 3种不同浓度OST预处理卵巢组织后再加入H2O2;同时设对照组。在显微镜下计数卵巢组织内卵泡总数和生长卵泡数，计算生长卵泡率;化学发光法检测培养液中雌二醇（E2）的水平，WST-1法检测卵巢组织中SOD活性，TBA法检测卵巢组织中MDA含量。结果 与对照组相比，H2O2损伤模型组卵巢生长卵泡数、卵泡总数和生长卵泡率均明显下降，培养第4天的培养液中E2浓度明显下降，组织内SOD活性显著下降（P<0.01），MDA含量明显升高（P<0.01）。40 μg/mL和80 μg/mL两个浓度的OST预处理后，卵巢组织中的生长卵泡数、卵泡总数和生长卵泡率、第4天培养液中的E2浓度、组织内SOD活性均较模型组有不同程度的上升，MDA含量下降（P<0.01）。20 μg/mL OST组仅在第4天培养液中的E2浓度上明显高于模型组，其他相关指标相对于模型组均无显著性差异。 结论 一定浓度的蛇床子素预处理可减轻H2O2对卵巢组织的氧化应激损伤。

[关键词] 蛇床子素;过氧化氢;氧化损伤;卵巢

[中图分类号] R285.5          [文献标识码] A          [文章编号] 1673-9701（2020）16-0035-05

Effects of Osthole on oxidative stress in ovarian tissue

LU Xiaosheng   WU Hao   XI Haitao   HUANG Zhaoxia   LV Jieqiang

Reproductive Center， the 2nd Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University， Wenzhou   325000， China

[Abstract] Objective To explore the protective effect of Osthole（OST） on ovarian oxidative damage. Methods Mice ovarian tissues were cultured in vitro. Hydrogen peroxide（H2O2） was used to construct oxidative damage models of ovarian tissues（model group）. The drug group was pretreated with different concentrations of OST， 20 μg/mL， 40 μg/mL and 80 μg/mL and H2O2 was added later; the control group was also set. The total number of follicles and the number of growing follicles in the ovarian tissue under a microscope was counted to calculate the rate of growing follicles; chemiluminescence method was applied to detect the level of estradiol（E2） in the culture fluid， WST-1 method to detect SOD activity in the ovarian tissue， and TBA method to detect the MDA content in ovarian tissue. Results Compared with the control group， the number of ovarian growth follicles， the total number of follicles， and the growth follicle rate in the H2O2 injury model group significantly reduced. The concentration of E2 in the culture fluid on the 4th day of culture significantly reduced， the SOD activity in the tissues significantly reduced（P<0.01）， and MDA content increased significantly（P<0.01）. After pretreatment with OST at 40 μg/mL and 80 μg/mL， the number of growing follicle in the ovarian tissue， the total number of follicles and the rate of growing follicles， the concentration of E2 in the culture medium on the 4th day， and the SOD activity in the tissues increased and MDA contents decreased as compared with the model group（P<0.01）. The E2 concentration of in the culture medium in 20 μg/mL OST group was significantly higher than that in the model group on the 4th day， and there were no significant differences in other related indicators compared with the model group. Conclusion Pretreatment with a certain concentration of OST can reduce the oxidative stress damage of ovarian tissue by H2O2.

三种不同浓度的OST组间比较，40 μg/mL和80 μg/mL两组在生长卵泡数上均明显高于20 μg/mL组（P<0.01），而40 μg/mL和80 μg/mL两组之间无统计学差异;3个不同浓度的OST组在卵泡总数上，组间均有统计学差异，以80 μg/mL组最高，20 μg/mL组最低（P<0.05）;3组之间在生长卵泡率上均无统计学差异（P>0.05），见表1、图1。以上结果提示20 μg/mL OST组的预处理不足以对H2O2在卵巢组织中造成的卵泡发育和构成上的损伤产生预防或缓解的作用，40 μg/mL OST组在卵泡数量与生长卵泡的比例上，较20 μg/mL OST组已有较大改善，而80 μg/mL OST组的浓度在本研究中对改善卵泡的总数和生长卵泡的比例均是最佳。

2.2 蛇床子素对体外培养卵巢E2分泌的影响

检测培养2 d收集的培养液中E2浓度，5组间比较差异均无统计学意义（P>0.05）;提示H2O2在培养的第2天尚未对卵巢的内分泌功能产生影响。

培养第4天，模型组在H2O2的作用下，培养液中E2的浓度明显下降，明显低于对照组（P<0.01），提示H2O2在培养第4天对卵巢的内分泌功能产生破坏;而3个不同浓度的OST组培养第4天的培养液E2浓度显著高于模型组（P<0.01），且与对照组相比无显著性差异。提示OST的预处理，可减缓H2O2对卵巢内分泌功能的影响。3个OST组之间比较亦无显著性差异（P>0.05），提示OST在较低浓度即可缓解H2O2对卵巢内分泌功能的影响，见表2、图2。

2.3 蛇床……