左归丸、右归丸复方及相关成分对小鼠胚胎干细胞1B10向生殖细胞方向分化的影响(1)
[摘要]目的：探索左归丸、右归丸及其相关成分对干细胞向生殖细胞分化的影响。方法：制备左归丸、右归丸、左归丸和右归丸共有成分的大鼠含药血清，以此作为实验药物;以小鼠胚胎干细胞1B10(MESC-1B10)为干细胞代表;然后以上述大鼠含药血清干预MESC-1B10，干预过程中进行显微照像追踪，干预72 h后，提取相关被干预细胞的RNA，立即合成cDNA，最后利用实时荧光定量PCR(qPCR)检测各干预细胞的10种与生殖分化相关基因的表达模式。结果：左归丸大鼠含药血清(ZGW-RS)显著上调了Oct-4和SCP3，显著下调了GDF-9和Stra8;右归丸大鼠含药血清(YGW-RS)显著上调了Oct-4，GDF-9，Mvh和SCP3，显著下调了Stra8，Itga6和Itgb1;左归丸和右归丸共有成分的大鼠含药血清(ZGWYGW-RS)显著上调了Oct-4，SCP3和ZP3，显著下调了GDF-9，Stra8，Itga6和Itgb1。结论：ZGW-RS能启动MESC-1B10向减数分裂方向变化，但是尚不足以推动1B10向生殖细胞分化;YGW-RS能启动MESC-1B10向减数分裂方向变化，同时推动1B10向雌性生殖细胞分化;ZGWYGW-RS能启动MESC-1B10向减数分裂方向变化，同时能轻微推动MESC-1B10向雌性生殖细胞分化，但是作用弱于YGW-RS。实验结果一方面增强了对左归丸和右归丸及其相关成分影响干细胞生殖分化的认识，另一方面有助于解释左归丸和右归丸治疗不孕不育症的机制。
, 百拇医药
[关键词]左归丸;右归丸;大鼠含药血清;小鼠胚胎干细胞;减数分裂;生殖分化相关基因
[收稿日期]2014-07-29
[基金项目]国家重点基础研究发展计划(973)项目 (2010Cb530403);成都中医药大学实验技术重点项目 (063003);成都中医药大学校基金项目(ZRMS201248)
[通信作者]*陆华，Tel：(028)85232577，E-mail：1298421480@qq.com
[作者简介]姚奏英，博士研究生，Tel：(028)85416273，E-mail：470030886@qq.comEffects of Zuogui pill， Yougui pill and relative compositions on differentiation
, 百拇医药
towards germ cells of mouse embryonic stem cell 1B10
YAO Zou-ying1，4， WAN Qian2，4 ， LU Hua3，4*， LIU Xia2，4
(1.The Affiliated College of Medical Sciences， Chengdu University of Traditional Chinese Medicine， Chengdu 610075， China;
2. The Second Affiliated College of Medical Sciences， Chengdu University of Traditional Chinese Medicine， Chengdu 610041， China;
3. The Second Affiliated Hospital， Chengdu University of Traditional Chinese Medicine， Chengdu 610041， China;
, 百拇医药
4. Sichuan Family Planning Research Institute， Chengdu 610041， China)
[Abstract]Objective： To investigate the effects of Zuogui pill， Yougui pill and the relative compositions on the differentiation towards germ cells of stem cells. Method： The rat drug sera for Zuogui pill， Yougui pill and the common composition of Zuogui pill and Yougui pill were prepared respectively as the experimental drugs; the mouse embryonic stem cell 1B10(MESC-1B10) was used as the representative of stem cells; the above rat drug sera were used to intervene MESC-1B10 and the process was traced by microscopy imaging; after 72 h of the intervention， the RNAs were extracted from the different intervened MESC-1B10， cDNAs were synthesized immediately and finally the Real-time quantitative PCR(qPCR) was used to measure the expression patterns of the 10 reproductive-differentiation-related genes for each intervention. Result： The rat drug serum of Zuogui pill(ZGW-RS) significantly up-regulated Oct-4 and SCP3 and significantly down-regulated GDF-9 and Stra8; the rat drug serum of Yougui pill (ZGW-RS) significantly up-regulated Oct-4， GDF-9， Mvh and SCP3 and significantly down-regulated Stra8， Itga6 and Itgb1; the rat drug sera for the common composition of Zuogui pill and Yougui pill (ZGWYGW-RS) significantly up-regulated Oct-4， SCP3 and ZP3 and significantly down-regulated GDF-9， Stra8， Itga6 and Itgb1. Conclusion： ZGW-RS can initiate the change towards meiosis， but can not start the reproductive differentiation of MESC-1B10; YGW-RS can initiate the change towards meiosis， and can also start the reproductive differentiation of MESC-1B10 towards female germ cells; ZGWYGW-RS can initiate the change towards meiosis， and can lightly start the reproductive differentiation of MESC-1B10 towards female germ cells but the inductive effect is smaller than YGW-RS. The experimental results， on one hand， strengthen the knowledge about the influence of the relative compositions of Zuogui pill and Yougui pill on the reproductive differentiation of stem cells， on the other hand， help to explain the mechanism of the treatment of the infertility by Zuogui pill and Yougui pill., 百拇医药(姚奏英 万谦 陆华 刘霞)
, 百拇医药
[关键词]左归丸;右归丸;大鼠含药血清;小鼠胚胎干细胞;减数分裂;生殖分化相关基因
[收稿日期]2014-07-29
[基金项目]国家重点基础研究发展计划(973)项目 (2010Cb530403);成都中医药大学实验技术重点项目 (063003);成都中医药大学校基金项目(ZRMS201248)
[通信作者]*陆华，Tel：(028)85232577，E-mail：1298421480@qq.com
[作者简介]姚奏英，博士研究生，Tel：(028)85416273，E-mail：470030886@qq.comEffects of Zuogui pill， Yougui pill and relative compositions on differentiation
, 百拇医药
towards germ cells of mouse embryonic stem cell 1B10
YAO Zou-ying1，4， WAN Qian2，4 ， LU Hua3，4*， LIU Xia2，4
(1.The Affiliated College of Medical Sciences， Chengdu University of Traditional Chinese Medicine， Chengdu 610075， China;
2. The Second Affiliated College of Medical Sciences， Chengdu University of Traditional Chinese Medicine， Chengdu 610041， China;
3. The Second Affiliated Hospital， Chengdu University of Traditional Chinese Medicine， Chengdu 610041， China;
, 百拇医药
4. Sichuan Family Planning Research Institute， Chengdu 610041， China)
[Abstract]Objective： To investigate the effects of Zuogui pill， Yougui pill and the relative compositions on the differentiation towards germ cells of stem cells. Method： The rat drug sera for Zuogui pill， Yougui pill and the common composition of Zuogui pill and Yougui pill were prepared respectively as the experimental drugs; the mouse embryonic stem cell 1B10(MESC-1B10) was used as the representative of stem cells; the above rat drug sera were used to intervene MESC-1B10 and the process was traced by microscopy imaging; after 72 h of the intervention， the RNAs were extracted from the different intervened MESC-1B10， cDNAs were synthesized immediately and finally the Real-time quantitative PCR(qPCR) was used to measure the expression patterns of the 10 reproductive-differentiation-related genes for each intervention. Result： The rat drug serum of Zuogui pill(ZGW-RS) significantly up-regulated Oct-4 and SCP3 and significantly down-regulated GDF-9 and Stra8; the rat drug serum of Yougui pill (ZGW-RS) significantly up-regulated Oct-4， GDF-9， Mvh and SCP3 and significantly down-regulated Stra8， Itga6 and Itgb1; the rat drug sera for the common composition of Zuogui pill and Yougui pill (ZGWYGW-RS) significantly up-regulated Oct-4， SCP3 and ZP3 and significantly down-regulated GDF-9， Stra8， Itga6 and Itgb1. Conclusion： ZGW-RS can initiate the change towards meiosis， but can not start the reproductive differentiation of MESC-1B10; YGW-RS can initiate the change towards meiosis， and can also start the reproductive differentiation of MESC-1B10 towards female germ cells; ZGWYGW-RS can initiate the change towards meiosis， and can lightly start the reproductive differentiation of MESC-1B10 towards female germ cells but the inductive effect is smaller than YGW-RS. The experimental results， on one hand， strengthen the knowledge about the influence of the relative compositions of Zuogui pill and Yougui pill on the reproductive differentiation of stem cells， on the other hand， help to explain the mechanism of the treatment of the infertility by Zuogui pill and Yougui pill., 百拇医药(姚奏英 万谦 陆华 刘霞)