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Effects of Dodder Extract-containing Serum on Apoptosis and Secretory Function of Isolated Mouse Ovarian Granulosa Cells |
ZHANG Jing-jing, XUE Bao-yao, ZHANG Ling-yan |
Department of Obstetrics and Gynelology, Shaanxi Provincial People's Hospital, Xi'an Shaanxi 710001 |
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Abstract 【Objective】To investigate the effects of serum containing dodder extract (semen cuscutae)-on apoptosis and secretion of isolated mouse ovarian granulosa cells in vitro. 【Methods】A total of 18 SPF-grade SD rats were divided into three groups (n=6). Rats in the low and high dose of dodder extract groups were intragastric administration of 893.77 mg/kg and 4468.55 mg/kg of Cuscuta Chinensis (semen cuscutae) for 3 consecutive days, respectively. Then serum with dodder extract and blank serum were obtained from abdominal aorta. Ten 3-week-old SPF SD female mice were intraperitoneally injected with pregnant equine serum gonadotropin 5 IU. Thereafter ovaries were isolated, granulosa cells were identified and cultured in vitro. The granulosa cells were further served as control group (cultured medium containing blank serum alone), ZEA group(medium containing 60 μmol/L ZEA and blank serum), low-dose and high-dose drug-containing serum group (medium containing 60 μmol/L ZEA plus low-dose and high-dose dodder extract-containing serum, respectively). The proliferation and apoptosis of granulosa cells were detected by CCK-8 and Annexinv/PITC staining and levels of Estradiol (E2) and progesterone (P) were detected by radioimmunoassay. Levels of PCNA, Bax-2 and Bcl-2 related x protein (Bax) were detected by immunoblot.【Results】Compared with the control group, the ovarian granulosa cell proliferation inhibition rate, apoptosis rate, Bax protein level increased (P<0.05), E2、P PCNA and Bcl-2 levels decreased(P<0.05) in ZEA group. Compared to ZEA group, the ovarian granulosa cell proliferation inhibition rate, apoptosis rate, Bax protein level in low and high dose of dodder extract-containing serum group decreased (P<0.05),while levels of E2,P,PCNA and Bcl-2 increased (P<0.05). The ovarian granulosa cell proliferation inhibition rate, apoptosis rate, Bax protein level in the high-dose dodder extract-containing serum group decreased more than those in the low-dose group(P<0.05), and levels of E2,P,PCNA andBcl-2 increased more than those in the low-dose group (P<0.05). 【Conclusion】The serum of dodder extract (semen cuscutae) can inhibit the apoptosis of ovarian granulosa cells induced by ZEA and improve the secretory function of ovarian granulosa cells.
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Received: 03 November 2021
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