Protective Effect and Mechanism of Proanthocyanidins B2 on Oxidative Damage in Porcine Granulosa Cells

doi:10.15933/j.cnki.1004-3268.2022.01.018

Journal of Henan Agricultural Sciences ›› 2022, Vol. 51 ›› Issue (1): 146-153.DOI: 10.15933/j.cnki.1004-3268.2022.01.018

• Animal Science Veterinary Medicine • Previous Articles Next Articles

Protective Effect and Mechanism of Proanthocyanidins B2 on Oxidative Damage in Porcine Granulosa Cells

XING Baosong¹，ZHANG Jiaqing¹，REN Qiaoling¹，LÜ Lingyan²，WANG Xianwei³，CHEN Junfeng¹，GAO Binwen¹，MA Qiang¹

（1.Institute of Animal Husbandry and Veterinary Science，Henan Academy of Agricultural Sciences/Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation，Zhengzhou 450002，China；2.Guangxi Institute of Animal Sciences，Nanning 530001，China；3.Henan Provincial Animal Husbandry General Station，Zhengzhou 450008，China）

Received:2021-04-30 Published:2022-01-15 Online:2022-03-18

原花青素B2 对猪颗粒细胞氧化损伤的保护作用及机制研究

邢宝松¹，张家庆¹，任巧玲¹，吕玲燕²，王献伟³，陈俊峰¹，高彬文¹，马强¹

（1.河南省农业科学院畜牧兽医研究所/河南省畜禽繁育与营养调控重点实验室，河南郑州 450002；2.广西壮族自治区畜牧研究所，广西南宁530001；3. 河南省畜牧总站，河南郑州 450008）

通讯作者: 张家庆（1982-），男，河北沧州人，助理研究员，博士，主要从事猪育种与繁殖研究。E-mail：zjq8650612@163.com。王献伟同为通信作者
作者简介:邢宝松（1969-），男，河南新密人，副研究员，主要从事猪育种与管理工作。E-mail：baosong@126.com
基金资助:
河南省自然科学基金项目（202300410529）；河南省农业科学院杰出青年科技基金项目（2021JQ07）；河南省农业科学院科技
创新创意项目（2020CX06）

Abstract

Abstract: In order to study the protective effect of proanthocyanidins B2（GSPB2）on oxidative damage of porcine granulosa cells（PGCs），the model of the damage on PGCs induced by H2O2 was made by primary culture.At the same time，different concentrations of GSPB2 were used to intervene.The cellular apoptosis was detected by TUNEL method. The changes of intracellular ROS were observed by fluorescence staining. The activities of superoxide dismutase（SOD），glutathione peroxidase（GSH‐Px），catalase（CAT）and the content of malondialdehyde（MDA）were measured by chemochromatometry methods. The apoptosis related genes，such as Bcl-2，Bax，Fas，Puma and Sirt1 were detected by quantitative PCR.The senescence related proteins，such as Sirt1，p16 and p21 were detected by Western blot.The results showed that the cell apoptosis and ROS level increased significantly，SOD，CAT and GSH‐Px activities decreased，and MDA content increased（P<0. 05）after treatment with 200 μmol/L H₂O₂ for 12 h. Furthermore，the 10 μmol/L GSPB2 significantly alleviated the above conditions induced by H₂O₂.Further research found that the expression ratio of Bcl-2/Bax was up‐regulated，Puma and Fas expression were down‐regulated，Sirt1 mRNA and protein expression was up‐regulated，and the protein expression of p16 and p21 was down‐regulated.The above results indicate that GSPB2 has a protective effect on oxidative damage in PGCs，which could be related with the up‐regulation of Sirt1 protein，down‐regulation of p16 and p21 protein expression，increase in Bcl-2/Bax ratio，and the inhibition of Fas and Puma expression.

Key words: Proanthocyanidins B2, Oxidative stress, Granulosa cell, Apoptosis

摘要： 为研究原花青素B2（GSPB2）对猪颗粒细胞氧化损伤的保护作用，采用原代细胞培养的方法，建立H2O2致颗粒细胞氧化损伤模型，同时用不同浓度GSPB2进行干预。采用TUNEL方法检测细胞凋亡，利用荧光染色法观察细胞内活性氧（ROS）变化，使用比色法检测细胞内超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化酶（GSH-Px）活性和丙二醛（MDA）含量，采用定量PCR方法检测凋亡相关基因Bcl-2、Bax、Fas、Puma、Sirt1 的mRNA表达水平，应用Western blot方法检测衰老相关分子Sirt1、p16和p21蛋白水平。结果显示，200 μmol/L H2O2处理12 h，可致细胞凋亡和ROS水平显著升高（P<0.05），SOD、CAT和GSH-Px活性下降，MDA 含量增加；10 μmol/L GSPB2 能显著减轻H₂O₂所致上述状况；且进一步研究发现，GSPB2抑制Fas 和Puma 的表达，提高Bcl-2/Bax 表达比例，上调Sirt1蛋白、下调p16和p21蛋白的表达。综上，GSPB2对猪颗粒细胞的氧化损伤具有保护作用，其机制可能与上调Sirt1蛋白、下调p16和p21蛋白表达，提高Bcl-2/Bax 比值，抑制Fas、Puma 表达有关。

关键词: 原花青素B2, 氧化应激, 颗粒细胞, 凋亡

CLC Number:

S828

XING Baosong, ZHANG Jiaqing, REN Qiaoling, LÜ Lingyan, WANG Xianwei, CHEN Junfeng, GAO Binwen, MA Qiang. Protective Effect and Mechanism of Proanthocyanidins B2 on Oxidative Damage in Porcine Granulosa Cells[J]. Journal of Henan Agricultural Sciences, 2022, 51(1): 146-153.

邢宝松, 张家庆, 任巧玲, 吕玲燕, 王献伟, 陈俊峰, 高彬文, 马强. 原花青素B2 对猪颗粒细胞氧化损伤的保护作用及机制研究[J]. 河南农业科学, 2022, 51(1): 146-153.

Figures/Tables 6

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Protective Effect and Mechanism of Proanthocyanidins B2 on Oxidative Damage in Porcine Granulosa Cells

原花青素B2 对猪颗粒细胞氧化损伤的保护作用及机制研究

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