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目的:探讨红景天苷诱导小鼠骨髓间充质干细胞marrow mesenchymal stem cells(MSCs)向神经元细胞定向分化的相关分子机制。方法:以MSCs(D1细胞)为对象,红景天苷作为诱导剂,运用流式细胞术、荧光免疫化学染色和Western blot方法分别检测红景天苷对细胞周期及Cyclin D1和p21蛋白表达的影响。结果:红景天苷诱导细胞24,48 h后,G0/G1期细胞比例升高,S期细胞比例下降,说明红景天苷能显著促进细胞发生G0/G1期阻滞;荧光免疫化学染色表明,对照组和诱导12~24 h后Cyclin D1为阳性表达,阳性细胞数为71%,71%和44%,而48~72 h组表达呈阴性;诱导12~72 h后p21呈阳性表达,阳性细胞数为85%,90%,65%和73%,与对照组相比差异显著(P<0.01);Western blot检测表明Cyclin D1表达量随诱导时间的延长下调,p21蛋白表达水平上调。结论:红景天苷通过影响细胞周期相关蛋白Cyclin D1和p21抑制细胞增殖,使MSCs阻滞于G0/G1期,进而促进MSCs定向分化。本试验为阐明红景天苷诱导MSCs定向分化的分子机制奠定了坚实的理论基础。
Objective: To investigate the molecular mechanism of salidroside-induced differentiation of mouse bone marrow mesenchymal stem cells (MSCs) into neurons. Methods: Using MSCs (D1 cells) as target, salidroside was used as an inducer to determine the cell cycle and expression of Cyclin D1 and p21 protein by salidroside using flow cytometry, fluorescence immunochemistry and Western blot. influences. RESULTS: Following 24 h and 48 h of cell salidroside induction, the proportion of G0/G1 phase cells increased and the proportion of S phase cells decreased, indicating that salidroside significantly promoted cell G0/G1 phase arrest; fluorescence immunochemical staining It was shown that Cyclin D1 was positively expressed in the control group and after 12-24 h of induction. The number of positive cells was 71%, 71%, and 44%, while it was negative in the 48-72 h group; p21 was positive after 12-72 h of induction. The number of positive cells was 85%, 90%, 65% and 73%, which was significantly different from that of the control group (P<0.01). Western blot showed that the expression of Cyclin D1 was down-regulated with the prolongation of induction time, and the expression of p21 protein was up-regulated. . CONCLUSION: Salidroside inhibits cell proliferation by influencing cell cycle-related proteins Cyclin D1 and p21, and arrests MSCs in G0/G1 phase, thereby promoting directional differentiation of MSCs. This experiment laid a solid theoretical basis for elucidating the molecular mechanism of salidroside-induced differentiation of MSCs.