【摘要】：目的:探討在體外造血微環(huán)境中肌源性干細胞Wnt/β-catenin信號通路相關(guān)基因的表達情況及當歸補血湯(DBD)載藥血清的干預作用。方法:將分離純化并經(jīng)流式細胞儀鑒定的MDSCs與BMSCs體外培養(yǎng),根據(jù)實驗需要隨機分為6組進行藥物干預,即空白對照組、共培養(yǎng)Wnt激動劑組、共培養(yǎng)Wnt抑制劑組、DBD+共培養(yǎng)組、DBD+共培養(yǎng)激動劑組及DBD+共培養(yǎng)抑制劑組。Real-time PCR檢測各實驗組MDSCs Wnt通路上下游節(jié)點基因Wnt3、Wnt3a、β-catenin及MDSCs增殖分化相關(guān)基因Cyclin D1、C-myc、CD34 mRNA的表達變化。結(jié)果:流式細胞儀檢測結(jié)果顯示MDSCs CD34、Sca-1陽性。倒置顯微鏡下觀察:共培養(yǎng)Wnt激動劑組、DBD+共培養(yǎng)組、DBD+共培養(yǎng)激動劑組MDSCs較空白對照組生長密集且生長速度快,激動劑組聚團明顯,共培養(yǎng)Wnt抑制劑組MDSCs增殖明顯受到抑制。Real-time PCR檢測結(jié)果顯示:DBD+共培養(yǎng)激動劑組的Wnt3、Cyclin D1、CD34 mRNA表達水平最高,共培養(yǎng)激動劑組與DBD共培養(yǎng)組其次,空白對照組最低。結(jié)論:當歸補血湯載藥血清作用于共培養(yǎng)體系下的肌源性干細胞,Wnt信號通路持續(xù)激活后,體外造血微環(huán)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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