[1]蔡 竞,李 楠,朱虹丽△,等.新加归肾丸通过调节mTOR信号通路促进体外培养大鼠卵巢颗粒细胞增殖*[J].陕西中医,2020,(9):1179-1183.[doi:DOI:10.3969/j.issn.1000-7369.2020.09.001]
 CAI Jing,LI Nan,ZHU Hongli,et al.Study on therapy of Xinjia Guishenwan of granulosa cells isolated from rat ovary and ecultured in vitro by regulating mTOR signal pathway[J].,2020,(9):1179-1183.[doi:DOI:10.3969/j.issn.1000-7369.2020.09.001]
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新加归肾丸通过调节mTOR信号通路促进体外培养大鼠卵巢颗粒细胞增殖*
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2020年9期
页码:
1179-1183
栏目:
基础研究
出版日期:
2020-09-05

文章信息/Info

Title:
Study on therapy of Xinjia Guishenwan of granulosa cells isolated from rat ovary and ecultured in vitro by regulating mTOR signal pathway
作者:
蔡 竞1李 楠2朱虹丽1△张 辉2李海英2
1.陕西中医药大学附属医院(咸阳 712021); 2.陕西中医药大学(咸阳 712046)
Author(s):
CAI JingLI NanZHU Hongliet al.
Affiliated Hospital of Shaanxi University of Chinese Medicine(Xianyang 712021)
关键词:
补肾活血法 新加归肾丸 mTOR信号通路 细胞周期 颗粒细胞增殖
Keywords:
Bushen Huoxue Xinjia Guishenwan mTOR signal pathway Cell cycle Proliferation of granulosa cell
分类号:
R711.7
DOI:
DOI:10.3969/j.issn.1000-7369.2020.09.001
文献标志码:
A
摘要:
目的:观察补肾活血中药复方新加归肾丸对体外培养大鼠卵巢颗粒细胞增殖的影响。方法:体外培养大鼠卵巢颗粒细胞,分为空白组,模型组,中药低、中、高剂量组共五组,空白组用1%胎牛血清培养基培养,其余四组滴加雷帕霉素,20 min后中药各组加入不同浓度的中药水提醇沉液。分别在培养24、48、72 h后采用流式细胞仪检测细胞周期各时相的变化,并计算细胞增殖指数(PI); 免疫印迹法检测磷酸化-哺乳动物雷帕霉素靶蛋白(p-mTOR)、细胞周期蛋白D2(Cyclin D2)的表达水平。结果:用药24、48、72 h 这3个时段,中药各组S期细胞比率、PI均高于模型组(P<0.05),其中中药中剂量组最为明显(P<0.05)。用药24、48、72 h这3个时段,中药各个剂量组p-mTOR、Cyclin D2表达水平均高于模型组(P<0.05)。中药各个剂量中以中药中剂量组效果最明显(P<0.05)。结论:补肾活血中药复方新加归肾丸能通过激活mTOR信号通路活性,上调Cyclin D2蛋白表达,促使颗粒细胞由G1期向S期转化,加速卵巢颗粒细胞周期进程,促进颗粒细胞增殖来实现改善卵泡发育。
Abstract:
Objective:To study the effect of Xinjia Guishenwan,compund Chinese medcine with the function of Bushen Huoxue,on proliferation of granulosa cells isolated from rat ovary and ecultured in vitro.Methods:Cultured granulosa cells isolated from rat ovary in vitro and divided them into blank groupmodel group,high-dosage TCM medicine group,mid-dosage TCM group,low-dosage TCM group.The blank group was cultured in 1% fetal bovine serum medium,the rest 4 groups were dripped with rapamycin for 20 minutes,then added water extracting-alcoholprecipitating solution of traditional Chinese medicine with different concentrations into TCM groups.After 24,48 and 72 hours of culture,detected the changes of each phase of granule cell cycle in each group by flow cytometry,and calculated the proliferation index(PI).The expression of p-mTOR、Cyclin D2 were detected by wstern blot.Results:At the 3 time periods(24 h,48 h,72 h)after therepy,the S-phase cells ratio and PI index of the TCM groups were all signifi-cantly higher than those of the model group(P<0.05).Among three TCM groups,the mid-dosage TCM group has the most obvious effect.At the 3 time periods(24 h,48 h,72 h)after therepy,the expression of mTOR and Cyclin D2 protein in the TCM groups were all significantly higher than those of the model group(P<0.05).The most evident effect was the mid-dosage TCM group.Conclusion:Compound Chinese medcine Xinjia Guishenwan for Bushen Huoxue improving can improve follicular development,maybe this effect is achieved by activating the activity of mTOR signal pathway and up-regulating the expression of Cyclin D2,induced the granulosa cell from G1 to S period,then accelerate cell cycle period and promote the prolifer-ation of granulosa cells.

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目(81603656);陕西省教育厅资助项目(14JK1198);陕西中医药大学学科创新团队项目(2019-QN03)
更新日期/Last Update: 2020-09-08