原花青素对玉米赤霉烯酮诱导牦牛颗粒细胞氧化损伤的保护作用研究

doi:10.11843/j.issn.0366-6964.2022.09.017

Abstract

Abstract: The aim of this study was to investigate the effects of procyanidins (PC) on the growth, proliferation, antioxidant properties and hormone secretion of yak granulosa cells after oxidative damage induced by zearalenone (ZEA). In this experiment, healthy yaks of 3-5 years old (n=3) were selected to complete the isolation and culture of ovarian granulosa cells, and immunofluorescence staining was performed to identify the purity of granulosa cells. The effects of different concentrations of ZEA (0 (Control group), 5, 10, 20, 40, 60, 80 and 100 μmol·mL^-1), different concentrations of PC (0 (Control group), 0.05, 0.5, 2.5, 5, 10, 50 and 100 μg·mL^-1) and the combined treatment of 50 μmol·mL^-1 ZEA+5 μg·mL^-1 PC on the activity of yak granulosa cells were compared by CCK-8 assay. The levels of reactive oxygen species (ROS) and estradiol (E₂) in granulosa cells of yaks Control group (without ZEA and PC), 50 μmol·mL^-1 ZEA and 50 μmol·mL^-1ZEA+5 μg·mL^-1PC groups were measured by ELISA. The expression levels of genes related to proliferation and growth, apoptosis, antioxidant and E₂ synthesis in different groups of granulosa cells were detected by real-time fluorescence quantitative PCR. The results showed that, cells isolated and cultured in this study expressed granulosa cell marker protein FSHR, which had high purity and could meet the requirements of subsequent experiments. After adding different concentrations of ZEA, the viability of granulosa cells decreased obviously with the increase of ZEA concentration (P<0.05). In a certain concentration range (0-5 μg·mL^-1), PC could significantly improve the viability of granulosa cells with the increase of concentration (P<0.05), and the effect was most obvious when the concentration was at 5 μg·mL^-1. Compared with ZEA treatment group, the combined treatment of ZEA+PC significantly increased the number of granulosa cells and cell viability(P<0.05). The expression of proliferation- and growth-related genes PCNA and IGF-II, as well as anti-apoptotic genes XIAP and BCL-2, significantly increased(P<0.05). On the contrary, the expression of pro-apoptotic genes BAX and CASP3 significantly decreased(P<0.05). Meanwhile, the combined treatment of ZEA+PC could significantly reduce the level of reactive oxygen species and promote the secretion of E₂ in yak granulosa cells(P<0.05). The expression of antioxidant-related genes SOD2, GPX1 and CAT, as well as E₂ synthesis-related genes STAR, CYP11A1 and HSD3B significantly up-regulated in granulosa cells(P<0.05). The above results suggest that PC promotes the growth and proliferation of granulosa cells after ZEA treatment, up-regulates the viability of granulosa cells, improves the antioxidant capacity, reduces the level of ROS, and increases the secretion level of E₂. In conclusion, PC has a protective effect on ZEA-induced oxidative damage in yak granulosa cells. This study provides some research data and theoretical support for the prevention and treatment of ZEA toxicity and the application of PC in animal husbandry.

Key words: zearalenone, proanthocyanidins, yak, granulosa cells, oxidative damage, estrogen

CLC Number:

S823.85

TANG Ziwen, CHENG Huaqin, LIU Dongju, PHAGMO Droma, YANG Xue, LI Jian, YIN Shi. The Protective Effect of Proanthocyanidins on Oxidative Damage of Yak Granulosa Cells Induced by Zearalenone[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3006-3017.

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The Protective Effect of Proanthocyanidins on Oxidative Damage of Yak Granulosa Cells Induced by Zearalenone

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