经当归补血汤干预的肌源性干细胞Wnt信号相关基因表达分析

发布时间：2019-05-14 05:57

【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。
[Abstract]:Aim: to investigate the expression of Wnt/ 尾-catenin signaling pathway related genes in muscle-derived stem cells in hematopoietic microenvironment in vitro and the intervention effect of Danggui Buxue decoction (DBD) loaded serum. Methods: MDSCs and BMSCs, which were isolated and purified and identified by flow cytometry, were randomly divided into 6 groups for drug intervention, namely, blank control group, co-culture Wnt agonist group, co-culture Wnt inhibitor group and DBD co-culture group. DBD co-culture agonist group and DBD co-culture inhibitor group. Real-time PCR detected the expression of MDSCs Wnt pathway upstream and downstream node genes Wnt3,Wnt3a, 尾-catenin and MDSCs proliferation and differentiation related genes Cyclin D1, C 鈮，

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