白藜芦醇拮抗卵泡颗粒细胞氧化应激的损伤

邢鹏; 张娜

doi:10.16098/j.issn.0529-1356.2020.02.007

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解剖学报 ›› 2020, Vol. 51 ›› Issue (2) : 189-194. DOI: 10.16098/j.issn.0529-1356.2020.02.007

细胞和分子生物学

白藜芦醇拮抗卵泡颗粒细胞氧化应激的损伤

邢鹏^1,2 张娜^1* 赵静云¹ 张轶¹ 吕翠婷¹许碧莎¹

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Antagonism of resveratrol on oxidative stress injury of follicular granulosa cells

XING Peng^1,2 ZHANG Na^1* ZHAO Jing-yun¹ZHANG Yi¹ Lü Cui-ting¹ XU Bi-sha¹

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摘要

目的探讨白藜芦醇是否可以拮抗过氧化氢诱导的卵泡颗粒细胞氧化应激损伤。方法收集体外受精/单精子卵泡浆内注射（IVF/ICSI）患者捡卵后卵泡液中的颗粒细胞并进行培养。将培养后的颗粒细胞分为4组：对照组，损伤模型组，10 μmol/L白藜芦醇组和50 μmol/L白藜芦醇组。采用CCK-8法测定细胞活性，硫代巴比妥酸（TBA）法测定丙二醛（MDA）含量，水溶性四氮唑-1（WST-1）法测定超氧化物歧化酶（SOD）水平，竞争ELISA 法测定孕酮分泌量，Annexin Ⅴ-FITC/PI双染流式细胞仪测定细胞凋亡，Western blotting测定Bcl-2、Caspase-3和沉默信息调节因子2相关酶1（SIRT1）蛋白表达。结果与对照组比较，损伤模型组的细胞活性、SOD水平、孕酮分泌量和Bcl-2蛋白表达均明显降低，而MDA含量、细胞凋亡率和Caspase-3凋亡蛋白表达均明显增加（P<0.05）。与损伤模型组比较，10 μmol/L白藜芦醇组各项指标未显示出统计学差异；而50 μmol/L白藜芦醇组的细胞活性、SOD水平、孕酮分泌量和Bcl-2及SIRT1蛋白表达均明显升高，MDA含量、细胞凋亡率和Caspase-3凋亡蛋白表达均明显下降（P<0.05）。结论 50 μmol/L白藜芦醇可以通过增加SIRT1的活性，增强颗粒细胞的抗氧化及抗凋亡能力，改善颗粒细胞的功能。

Abstract

Objctive To investigate the protective effect of resveratrol on oxidative stress damage of follicular granulosa cells induced by hydrogen peroxide. Methods Granulosa cells were collected from the follicular fluid of in vitro fertilization(IVF) patients after oocyte retrieval and cultured. The cultured granulosa cells were divided into four groups: control group, injury model group, 10 μmol/L resveratrol group and 50 μmol/L resveratrol group. Cell viability was determined by CCK-8 assay, malondialdehyde(MDA) content by thiobarbituric(TBA) assay, superoxide dismutase(SOD) level by water soluble tetrazdium-1(WST-1) assay, apoptosis by AnnexinV-FITC/PI double-staining flow cytometry, Bcl-2 and Caspase-3 protein expression by Western blotting, and progesterone secretion by competitive ELISA. Resutls Compared with the control group, the cell viability, SOD level, Bcl-2 protein expression and progesterone secretion were significantly decreased in the injury model group, while MDA content, apoptosis rate and Caspase-3 apoptotic protein expression were significantly increased (P<0.05). Compared with the injury model group, the 10 μmol/L resveratrol group showed no statistically significant differences in various parameters; however, the cell viability, SOD level, progesterone secretion, and Bcl-2 and silent information regulator factor 2 related enzyme 1(SIRT1) protein expression were significantly increased, and the MDA content, apoptosis rate, and Caspase-3 apoptotic protein expression were significantly decreased in the 50 μmol/L resveratrol group (P<0.05). Conclusion 50 μmol/L resveratrol can increase the activity of SIRT1, enhance the anti-oxidation and anti-apoptosis ability of granulosa cells and improve the function of granulosa cells.

关键词

白藜芦醇 / 过氧化氢 / 卵泡颗粒细胞 / 氧化应激 / 抗氧化 / 免疫印迹法 / 人

Key words

Resveratrol

/ Hydrogen peroxide / Follicular granulosa cells / Oxidative stress / Antioxidants / Western blotting / Human

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邢鹏张娜赵静云张轶吕翠婷许碧莎. 白藜芦醇拮抗卵泡颗粒细胞氧化应激的损伤[J]. 解剖学报. 2020, 51(2): 189-194 https://doi.org/10.16098/j.issn.0529-1356.2020.02.007

XING Peng ZHANG Na ZHAO Jing-yun ZHANG Yi Lü Cui-ting XU Bi-sha. Antagonism of resveratrol on oxidative stress injury of follicular granulosa cells[J]. Acta Anatomica Sinica. 2020, 51(2): 189-194 https://doi.org/10.16098/j.issn.0529-1356.2020.02.007

中图分类号： R711.6

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