摘要
目的:探讨在体外造血微环境中肌源性干细胞Wnt/β-catenin信号通路相关基因的表达情况及当归补血汤(DBD)载药血清的干预作用。方法:将分离纯化并经流式细胞仪鉴定的MDSCs与BMSCs体外培养,根据实验需要随机分为6组进行药物干预,即空白对照组、共培养Wnt激动剂组、共培养Wnt抑制剂组、DBD+共培养组、DBD+共培养激动剂组及DBD+共培养抑制剂组。Real-time PCR检测各实验组MDSCs Wnt通路上下游节点基因Wnt3、Wnt3a、β-catenin及MDSCs增殖分化相关基因Cyclin D1、C-myc、CD34 mRNA的表达变化。结果:流式细胞仪检测结果显示MDSCs CD34、Sca-1阳性。倒置显微镜下观察:共培养Wnt激动剂组、DBD+共培养组、DBD+共培养激动剂组MDSCs较空白对照组生长密集且生长速度快,激动剂组聚团明显,共培养Wnt抑制剂组MDSCs增殖明显受到抑制。Real-time PCR检测结果显示:DBD+共培养激动剂组的Wnt3、Cyclin D1、CD34 mRNA表达水平最高,共培养激动剂组与DBD共培养组其次,空白对照组最低。结论:当归补血汤载药血清作用于共培养体系下的肌源性干细胞,Wnt信号通路持续激活后,体外造血微环境中的MDSCs更易增殖并分化。
Objective：To investigate the variation of gene expression of Wnt signaling pathway on muscle derived stem cells in- tervene by Danggui Buxue Decoction（DBD） in vitro hematopoietic microenvironment. Methods：MDSCs purified and detected by FACS and co - cultured with BMSCs in vitro were randomly divided into 6 groups. The expressions of Wnt3,Wnt3a, [3 - eatenin, CD34,C - mye and CyclinD1 mRNA in MDSCs were detected by RT - PCR and the expressions of CD34 and Sca- 1 in MDSCs were detected by FACS. Results ：The FACS showed that CD34 and Sca - 1 were positive in MDSCs. Observed under the inverted microscope, MDSCs in co - cultured Wnt agitated group, DBD ＋ co - cultured group, DBD ＋ co - cultured Wnt agitated group mat- ted growth faster than those in blank control group. MDSCs in DBD ＋ Wnt agitated group was clustered obviously. The proliferation of MDSCs was significantly suppressed in co - cultured Wnt inhibited group. RT - PCR analysis showed that the expressions of Wnt3, CyclinD1, CD34 mRNA in DBD ＋ co - cultured Wnt agitated group were the highest and that in co - cultured Wnt agitated group were higher than those in blank control group. Conclusion：The sustained activation of Wnt signaling pathway can promote the proliferation and differentiation of MDSCs intervened by Danggui Buxue Decoction in vitro hematopoietic microenvironment.
出处
《辽宁中医杂志》
CAS
北大核心
2017年第12期2486-2489,共4页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81673732
81503401
81273892)
国家大学生创新创业计划项目(201510063006)
关键词
肌源性干细胞
WNT信号通路
当归补血汤
造血微环境
muscle derived stem cells （MDSCs）
Wnt singling pathway
Danggui Buxue Decoction
hematopoietic microenvi- ronment
