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目的筛选补肾活血汤促进大鼠骨髓间充质干细胞(BMSCs)体外迁移的活性部位并研究其对CXCR4蛋白表达的影响。方法补肾活血汤采用索氏提取法,得到石油醚、乙酸乙酯、无水乙醇及水提物4个部位。全骨髓贴壁培养法培养BMSCs,取第三代(P3)BMSCs进行实验。补肾活血汤4个部位按照10,50,100μg·mL~(-1)浓度梯度对BMSCs进行Transwell实验,筛选出有效部位。使用Western blot和细胞免疫荧光检测补肾活血汤有效部位对CXCR4蛋白表达的影响。结果 Transwell结果显示,补肾活血汤石油醚部位具有明显促进大鼠BMSCs体外迁移的作用,以100μg·mL~(-1)浓度最佳(P<0.05)。Western blot、细胞免疫荧光结果显示,补肾活血汤石油醚部位呈浓度依赖性上调大鼠BMSCs的CXCR4蛋白表达(P<0.05)。结论补肾活血汤促进大鼠BMSCs体外迁移的有效部位为石油醚部位,其相关机制可能与上调CXCR4蛋白表达,激活SDF-1/CXCR4信号轴有关。
Objective To screen the role of Bushen Huoxue Decoction in promoting the migration of rat bone marrow mesenchymal stem cells (BMSCs) in vitro and to study its effect on CXCR4 protein expression. Methods Bushen Huoxue Decoction with Soxhlet extraction method, petroleum ether, ethyl acetate, ethanol and water extract 4 parts. BMSCs were cultured in the whole bone marrow adherent culture method and the third generation (P3) BMSCs were used in the experiment. Bushen Huoxue Decoction (BNSCs) was transwell assayed in 4 sites according to the concentration gradient of 10, 50 and 100μg · mL ~ (-1), and the effective sites were screened out. Western blot and immunofluorescence were used to detect the effect of Bushen Huoxue Decoction on CXCR4 protein expression. Results The results of Transwell showed that the fraction of petroleum ether of Bushen Huoxue Decoction significantly promoted the in vitro migration of rat BMSCs with the best concentration of 100 μg · mL -1 (P <0.05). Western blot and immunofluorescence showed that the petroleum ether fraction of Bushen Huoxue Decoction up-regulated the CXCR4 protein expression in BMSCs in a dose-dependent manner (P <0.05). Conclusion Bushen Huoxue Decoction can promote the migration of BMSCs in vitro. It is related to the up-regulation of CXCR4 protein expression and activation of SDF-1 / CXCR4 signal axis.