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| Effects of Naringenin on Proliferation, Apoptosis and Estradiol Synthesis of Granulosa cells in Tibetan Sheep (Ovis aries) |
| LIN Qian1, WANG Li-Bin1,2,*, ZHAO Bo1, WANG Meng1,2, PAN Yang-Yang1,2, JIAO Zheng-Xing1, XU Rui-Hua1, ZHANG Hui1, YU Si-Jiu1,2 |
1 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
2 Technology and Research Center of Gansu Province for Embryonic Engineering of Livestock, Lanzhou 730070, China |
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Abstract As the main economic animal in the Qinghai-Tibet plateau of China, the development of Tibetan sheep (Ovis aries) population is limited due to its weak reproductive ability, low conception rate and low pregnancy rate. Therefore, it is of great significance to study the problems related to Tibetan sheep reproduction. This study was designed to investigate the effects of naringenin (NAR) on proliferation, apoptosis and estradiol (E2) synthesis of granulosa cells (GCs) in Tibetan sheep. The primary GCs of Tibetan sheep were collected and cultured in vitro to logarithmic growth phase, different concentrations of NAR (0, 5, 10, 15 and 20 μmol/L) were added to culture GCs for 12 h. qPCR and Western blot were used to detect the expressions of genes and proteins related to proliferation (PCNA, CCND1), apoptosis (Bcl-2, BAX, Caspase-3) and E2 synthesis (CYP19A1, CYP17A1, CYP11A1, STAR). The proliferation of GCs was detected by CCK-8, and the apoptosis of GCs was detected by immunofluorescence. The concentration of E2 in the culture medium was detected by enzyme-linked immunosorbent assay (ELISA). The results showed that adding 10 μmol/L of NAR significantly up-regulated the expression of CCND1 mRNA (P<0.05). And the expressions of proliferation-related proteins (PCNA, CCND1) in NAR treatment groups were significantly higher than those of the control group (P<0.05). The expressions of genes and proteins of BAX and Caspase3 in the treatment groups decreased significantly (P<0.05). The expression levels of E2 synthesis-related genes and proteins (CYP11A1, CYP17A1 and CYP19A1) in the 10 μmol/L NAR treatment group rose significantly (P<0.05), while those of STAR mRNA and protein decreased significantly (P<0.05). In summary, NAR of 10 μmol/L promoted the proliferation of GCs and inhibited their apoptosis by up-regulating PCNA, CCND1, Bcl-2 mRNA and down-regulating BAX and Caspase3 protein expression; E2 was synthesized and secreted by up-regulating CYP11A1, CYP17A1, CYP19A1 and down-regulating STAR mRNA and proteins. This study provides a foundation for further study on the effects of NAR on functions of mammalian ovaries and improvement the animal reproductive potential.
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Received: 09 December 2024
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Corresponding Authors:
*wanglb@gsau.edu.cn
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