Protective Effect of Lycopene Against H2O2-induced Oxidative Damage in Porcine Ovarian Granulosa Cells
YANG Zhen-Qing1,2,3, HU Guang-Ling1,2,3, SHI Qing-Wu1,2,3, LI Chang-Hong4, CHEN Guang4, WANG Jie1,2,3, ZHANG Yi-Yu1,2,3, AO Zheng1,2,3,*
1 Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guiyang 550025, China; 2 Guizhou Provincial Key Laboratory of Animal Genetics, Breeding and Reproduction, Guiyang 550025, China; 3 College of Animal Science, Guizhou University, Guiyang 550025, China; 4 Guizhou Fuzhiyuan Agricultural Development Co., Ltd., Guiyang 550009, China
Abstract：The ovaries of sows (Sus scrofa) suffer oxidative stress can affect their reproductive performances. Lycopene (LYC), a natural carotenoid, has strong antioxidant properties and effectively alleviate oxidative damage in the body.The aim of this work was to investigate the protective effect of LYC against hydrogen peroxide (H2O2)-induced apoptosis and oxidative damage in porcine ovarian granulosa cells (PGCs). In this study, healthy and sexually mature porcine ovarian tissues were collected to isolate and culture PGCs, dividing the experiment into 3 groups: The control group, the H2O2 model group, and the LYC+H2O2 treatment group. Cell counting kit-8 (CCK-8) was used to detect cell viability to screen the optimal concentration of H2O2 damage and LYC treatment, and the kits were used to detect ROS and related antioxidant indexes. qRT-PCR was used to detect the mRNA expression levels of the genes related to cell apoptosis B cell lymphoma-2 (BCL-2), BCL-2-associated X protein (BAX), cysteinyl aspartate-specific proteinase-3 (CASP3), cell proliferation proliferating cell nuclear antigen (PCNA), insulin-like growth factor 2 (IGF2), antioxidant enzyme system catalase (CAT), manganese superoxide dismutase (MnSOD), copper-zinc superoxide dismutase (CuZnSOD), and steroid hormone biosynthesis steroidogenic acute regulatory protein (STAR), 3β-hydroxysteroid dehydrogenase 1 (HSD3B1), cytochrome P450 11A1 (CYP11A1), cytochrome P450arom (CYP19A1). Results indicated that 5 μmol/L LYC was most effective in promoting PGC viability, while 200 μmol/L H2O2 represented the IC50 for PGCs. Compared to the control group, the H2O2 model group showed extremely significantly elevated intracellular ROS fluorescence, ROS levels, and malondialdehyde (MDA) content (P<0.01), alongside decreased levels of total superoxide dismutase (T-SOD), total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), and catalase (CAT) (P<0.05). The mRNA expression levels of BCL-2, PCNA, IGF2, CuZnSOD, MnSOD, CAT, STAR, HSD3B1, CYP11A1and CYP19A1 were extremely significantly downregulated in the H?O? model group (P<0.01), while BAX, CASP3 were extremely significantly upregulated (P<0.01) compared with the control group. In the LYC + H?O? group, intracellular ROS fluorescence, ROS levels, and MDA content extremely significantly decreased compared to the H?O? model group (P<0.01), whereas cell viability, T-SOD, T-AOC, GSH-Px, and CAT levels extremely significantly increased (P<0.01). Additionally, LYC treatment significantly upregulated the mRNA expression of PCNA, IGF2, BCL-2, CuZnSOD, MnSOD, CAT, STAR, CYP11A1, and CYP19A1 (P<0.05) and significantly downregulated BAX and CASP3 (P<0.05) compared with the H2O2 model group. These results suggested that LYC had a protective effect on porcine ovarian granulosa cells by alleviating oxidative stress damage caused by H2O2. This study provides theoretical basis for the application of LYC in the swine industry.
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