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Yangjing Capsule Extract Promotes Proliferation of GC-1Spg Cells via Up-regulated pou3f1 Pathway

Yangjing Capsule Extract Promotes Proliferation of GC-1Spg C...

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第十次全国中西医结合男科学术大会、第六次广西中医、中西医结合男科学术大会、全国中西医结合男科疾病诊疗新进展学习班

作者： Bin Cai Baofang Jin Xindong Zhang Yuegui Cui Dalin Sun Bin Chen Guanghui Chen Yong Wang Chao Gao Andrology Department of Integrative Medicine Zhongda HospitalSoutheast University Reproductive Medicine Center the Affiliated Drum Tower HospitalMedical CollegeNanjing University State Key Laboratory of Reproductive Medicine Clinical Center of Reproductive MedicineFirst Affiliated HospitalNanjing Medical University Department of Traditional Chinese Medicine People’s Hospital of Suzhou Gaoxin District

Background:As similar to glial cell line-derived neurotrophic factor(GDNf),the Yangjing Capsule(YC)extract could also lead to proliferation of spermatogonial stem cells(SSCs)by motivating phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathway;however,the regulatory effect of YC extract on the expression of pou3f1still remains ***:To determine whether the transcription factor pou3f1 is up-regulated by YC extract through PI3K/AKT signaling pathway to regulate SSCs survival and ***:The cultured GC-1 spermatogonial(spg)cells were treated with 0.01,0.1,and 1 mg/mL YC extract for 48 *** cell viability was analyzed using MTT assay,while pou3f1 expression was quantitatively detected using real-time polymerase chain reaction and Western blot ***3f1,GDNf family receptor alphal(GfRα1)short interfering ribonucleic acid(siRNA),and LY294002(PI3K inhibitor)were applied as blockers to explore the underlying ***:After 48 h treatment with YC extract,the GC-1 spg cells were proliferatedand the pou3f1 expression was significantly increased in a dose-dependent *** pou3f1 siRNA partially blocked those effects of YC *** GfRα1siRNA and LY294002,as upstream blockers,reduced the pou3f1 expression induced by YC ***:The YC extract promotes proliferation of GC-1 spg cells via up-regulation of *** potential mechanism is that YC extract triggers the activation of PI3K/AKT pathway and then up-regulates pou3f1 expression.

关键词： Yangjing Capsule(YC) glial cell line-derived neurotrophic factor(GDNf) spermatogonial stem cells(SSCs) pou3f1

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Yangjing Capsule Extract Promotes Proliferation of GC-1 Spg Cells via Up-regulation of pou3f1

Yangjing Capsule Extract Promotes Proliferation of GC-1 Spg ...

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中华中医药学会第十四次男科学术大会

作者： Bin Cai Baofang Jin Xindong Zhang Dalin Sun Yugui Cui Chao Gao Institute of Andrology Nanjing University of Chinese Medicine State Key Laboratory of Reproductive Medicine Clinical Center of Reproductive MedicineFirst Affiliated HospitalNanjing Medical University

Objective:To investigate the role of pou3f1 in proliferation of GC-I spermatogonia(spg)cells cultured by Yangjing Capsule(YC)***:GC-1 spg cells were treated with0.01,0.1,and 1 mg/mL YC ***-8 assay was performed to detect the cell *** cytometry was used to measure the cell cycle and apoptosis of GC-1 spg ***-time PCR and westem blot were applied to determine the mRNA and protein expression of ***α1 and pou3f1 knockdown and LY294002(PI3K inhibitor)were applied to explore the underlying ***:After 48 h treatment of YC,the viability of GC-1 spg cells increased significantly and the ratio of apoptotic cells reduced *** increased mRNA and protein expression of pou3f1 and down-regulation of it blocked by pou3f1 siRNAs suggested YC promoted self-renewal of GC-1 spg cells via Up-regulation of *** Gfrα1 siRNAs andLY294002 treatments held back YC extract’s stimulation effects on mRNA and protein expression of pou3f1 and consequently inhibited the proliferation of GC-1 spg cells induced by YC ***:YC extract could stimulate the proliferation of GC-1 spg *** extract is able to trigger the activation of PI3K pathway and up-regulates the expression of pou3f1downstreamand finally leads to self-renewal ofGC-1 spg cells.

关键词： Yangjing capsule GC-1 pou3f1 Proliferation PI3K

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养精胶囊提取液通过上调pou3f1促进GC-1精原干细胞增殖的机制研究

养精胶囊提取液通过上调POU3F1促进GC-1精原干细胞增殖的机制研究

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作者：蔡滨南京中医药大学

学位级别：硕士

[目的]观察养精胶囊是否通过上调pou3f1促进GC-1细胞的增殖,并探讨其具体的信号通路机制。[方法]以体外小鼠精原细胞系GC-1细胞培养为模型,用无血清培养液诱导GC-1细胞凋亡,继而加入养精胶囊提取液(0.01,0.1, 1mg/mL)作用48小时后,用MT... 详细信息

[目的]观察养精胶囊是否通过上调pou3f1促进GC-1细胞的增殖,并探讨其具体的信号通路机制。[方法]以体外小鼠精原细胞系GC-1细胞培养为模型,用无血清培养液诱导GC-1细胞凋亡,继而加入养精胶囊提取液(0.01,0.1, 1mg/mL)作用48小时后,用MTT法检测细胞的活性;实时荧光定量PCR法和Western blot法检测pou3f1的mRNA和蛋白质的表达;利用pou3f1 siRNA, GfRal siRNA和LY294002分别阻断GDNf-PI3K/Akt-pou3f 1通路的上下游,探究其作用机制。[结果]养精胶囊提取液加入GC-1细胞后,能够显著增加细胞的活力,pou3f1的mRNA和蛋白质的表达水平增加;使用pou3f1 siRNA, GfRα1 siRNA和LY294002处理后,明显抑制了养精胶囊提取液对GC-1细胞的增殖,降低了养精胶囊提取液对pou3f1的mRNA和蛋白质的表达。[结论]养精胶囊提取液能促进GC-1细胞的增殖。通过结合GfRα1,养精胶囊提取液能引发PI3K/AKT通路的激活,上调pou3f1的表达,最终导致GC-1细胞的增殖。

关键词：养精胶囊 GDNf 精原干细胞 pou3f1

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