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目的探讨丹酚酸B(SalB)对骨髓间充质干细胞(MSC)存活的影响及其作用机制。方法体外培养MSC,建立体外缺血缺氧(OGD)模型。实验分MSC组、MSC+OGD组和MSC+OGD+不同浓度丹酚酸B组,MSC+OGD+丹酚酸B组在进行OGD处理前30 min,加入相应浓度的SalB进行预处理。通过Hochest33342染色和Annexin-V/PI双重染色流式细胞仪检测细胞凋亡和存活,经Rhodamine 123染色观察细胞线粒体膜电位的变化,通过Western blot检测Bcl-2和Bax的表达。结果 Hochest33342染色和Annexin-V/PI流式细胞仪检测显示,MSC经OGD处理后,出现明显的凋亡现象(P<0.01)。OGD处理前加入SalB与单纯OGD处理比较,MSC的凋亡明显减少,存活细胞数目显著增多(P<0.05,P<0.01)。OGD处理可以降低Rhodamine 123的表达,减少Bcl-2水平、增加Bax水平(P<0.01);与OGD组比较,10μmol/L丹酚酸B预处理可增加Rhodamine 123的表达,并可增加Bcl-2水平、减少Bax水平(P<0.05)。结论 SalB预处理可阻止线粒体凋亡途径,从而减少MSC凋亡,提高干细胞的存活率。
Objective To investigate the effect of salvianolic acid B (SalB) on the survival of bone marrow mesenchymal stem cells (MSCs) and its mechanism. Methods MSC were cultured in vitro to establish an in vitro hypoxic-ischemic (OGD) model. The experimental groups were divided into MSC group, MSC + OGD group and MSC + OGD + salvianolic acid B group, MSC + OGD + Salvianolic acid group B for 30 min before OGD treatment, and SalB pretreatment with corresponding concentration. Cell apoptosis and survival were detected by Hochest33342 staining and Annexin-V / PI double staining flow cytometry. Rhodamine 123 staining was used to observe the change of mitochondrial membrane potential. Western blot was used to detect the expression of Bcl-2 and Bax. Results Hochest33342 staining and Annexin-V / PI flow cytometry showed that after OGD treatment, obvious apoptosis was observed (P <0.01). Compared with OGD treatment, the apoptosis of MSC was significantly decreased and the number of viable cells was significantly increased (P <0.05, P <0.01). OGD could decrease Rhodamine 123 expression, decrease Bcl-2 and increase Bax (P <0.01). Compared with OGD group, pretreatment with 10 μmol / L salvianolic acid B increased Rhodamine 123 expression and increased Bcl- 2 levels, reducing Bax levels (P <0.05). Conclusion SalB pretreatment can block the mitochondrial apoptosis pathway, thereby reducing MSC apoptosis and increasing the survival rate of stem cells.