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医学临床研究  2022, Vol. 39 Issue (1): 55-58    DOI: 10.3969/j.issn.1671-7171.2022.01.015
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菟丝子提取物含药血清对离体小鼠卵巢颗粒细胞凋亡及分泌功能的影响
张京京, 薛宝瑶, 张凌燕*
陕西省人民医院妇产科,陕西 西安 710001
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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摘要 【目的】探讨菟丝子提取物含药血清对离体小鼠卵巢颗粒细胞凋亡及分泌功能的影响。【方法】各取6只SPF级雌性SD大鼠同时间分别灌胃菟丝子提取物溶液893.77 mg/kg、4468.85 mg/kg、等量生理盐水,连续3 d给药,3 d后取腹主动脉血分别得低、高剂量含药血清及空白血清;10只3周龄SPF级SD雌性小鼠每只腹腔注射孕马血清促性腺激素5 IU,分离卵巢并分离、鉴定、培养小鼠卵巢颗粒细胞。将细胞分为空白对照组(加入含空白血清的培养基培养);玉米赤霉烯酮(ZEA)组(加入含ZEA终浓度为60 μmol/L及空白血清的培养基);低、高剂量含药血清组(分别加入ZEA终浓度为60 μmol/L及低、高剂量含药血清的培养基)。采用CCK-8法、AnnexinV/PITC双染法分别检测小鼠卵巢颗粒细胞增殖、凋亡情况;采用放射免疫法检测小鼠卵巢颗粒细胞分泌雌二醇(E2)、孕酮(P)水平;免疫印迹法检测卵巢颗粒细胞增殖、凋亡蛋白增殖细胞核抗原(PCNA)、B淋巴细胞瘤基因-2(Bcl-2)、Bcl-2相关X蛋白(Bax)水平。【结果】与空白对照组比较,ZEA组卵巢颗粒细胞增殖抑制率、凋亡率、Bax蛋白水平升高(P<0.05),PCNA、Bcl-2蛋白水平及分泌E2、P水平降低(P<0.05);与ZEA组比较,低、高剂量含药血清组卵巢颗粒细胞增殖抑制率、凋亡率、Bax蛋白水平呈剂量依赖性降低(P<0.05),PCNA、Bcl-2蛋白及E2、P水平呈剂量依赖性升高(P<0.05)。【结论】菟丝子提取物含药血清能够抑制ZEA诱导的卵巢颗粒细胞凋亡,并提高卵巢颗粒细胞的分泌功能。
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张京京
薛宝瑶
张凌燕
关键词 植物提取物/药理学卵丘细胞小鼠疾病模型,动物细胞凋亡    
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.
Key wordsPlant Extracts/PD    Cumulus Cells    Mice    Disease Models, Animal    Apoptosis
收稿日期: 2021-11-03     
中图分类号:  R339.221  
通讯作者: *E-mail:zlingyan1984@163.com   
引用本文:   
张京京, 薛宝瑶, 张凌燕. 菟丝子提取物含药血清对离体小鼠卵巢颗粒细胞凋亡及分泌功能的影响[J]. 医学临床研究, 2022, 39(1): 55-58.
ZHANG Jing-jing, XUE Bao-yao, ZHANG Ling-yan. Effects of Dodder Extract-containing Serum on Apoptosis and Secretory Function of Isolated Mouse Ovarian Granulosa Cells. JOURNAL OF CLINICAL RESEARCH, 2022, 39(1): 55-58.
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http://journal07.magtech.org.cn/yxlcyj/CN/10.3969/j.issn.1671-7171.2022.01.015     或     http://journal07.magtech.org.cn/yxlcyj/CN/Y2022/V39/I1/55
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