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目的:探讨补肾活血汤对骨折端间充质干细胞(Mesenchymal Stem Cells,MSCs)体外迁移及表面受体CC类趋化因子受体2(C-C motif chemokine receptor-2,CCR2)表达的影响。方法:补肾活血汤灌胃治疗大鼠股骨中段骨折模型7、14、21d,分别取骨折端MSCs进行培养,传至第3代采用Transwell体外迁移实验检测补肾活血汤对骨折端MSCs体外迁移的影响,Western Blot、qPCR检测CCR2的表达。结果:补肾活血汤治疗后第7、14天,骨折端MSCs体外迁移能力明显加强,与模型组、对照组比较,差异有统计学意义(P<0.05)。第21天,骨折端MSCs体外迁移能力明显减弱,与模型组比较,差异无统计学意义(P>0.05)。Western blot、qPCR结果显示,补肾活血汤治疗第7天,治疗组CCR2的表达明显高于模型组、对照组(P<0.05)。第14天,CCR2的表达略有上升,与治疗后7d比较,差异无统计学意义(P>0.05),但治疗组仍显著高于模型组、对照组(P<0.05)。第21天,CCR2的表达有所下降,与治疗后7d比较,差异有统计学意义(P<0.05)。但治疗组与模型组比较,差异无统计学意义(P>0.05)。实验结果还表明,对照组CCR2的表达均处于低水平,7、14、21d比较,差异无统计学意义(P>0.05)。结论:补肾活血汤可以促进大鼠骨折愈合过程中MSCs的体外迁移,其相关机制可能与上调CCR2的表达、激活MCP-1/CCR2信号轴有关。
Objective: To investigate the effect of Bushen Huoxue decoction on the migration of Mesenchymal Stem Cells (MSCs) in vitro and the expression of CCR2 on the surface of fracture. Methods: Bushen Huoxue Decoction was used to treat middle femur fractures model in rats for 7, 14, and 21 days. MSCs were harvested for osteogenesis at the third passage. Transwell migration assay was used to determine the effect of Bushen Huoxue Decoction on the migration of MSCs in vitro , Western Blot, qPCR detection of CCR2 expression. Results: On the 7th and 14th day after the treatment of Bushen Huoxue Decoction, MSCs migrated in vitro significantly enhanced in vitro. Compared with the model group and the control group, the difference was statistically significant (P <0.05). On the 21st day, MSCs migrated in vitro significantly reduced in vitro, with no significant difference compared with the model group (P> 0.05). Western blot and qPCR showed that on the 7th day of treatment with Bushen Huoxue Decoction, the expression of CCR2 in the treatment group was significantly higher than that in the model group and the control group (P <0.05). On the 14th day, the expression of CCR2 slightly increased, but there was no significant difference compared with the 7th day after treatment (P> 0.05). However, the treatment group was still significantly higher than the model group and the control group (P <0.05). On the 21st day, the expression of CCR2 was decreased, compared with the 7th day after treatment, the difference was statistically significant (P <0.05). However, the treatment group and the model group, the difference was not statistically significant (P> 0.05). The experimental results also showed that the expression of CCR2 in the control group was at a low level, with no significant difference at 7, 14 and 21 days (P> 0.05). Conclusion: Bushen Huoxue Decoction can promote the migration of MSCs in vitro during fracture healing. The mechanism may be related to the up-regulation of CCR2 expression and the activation of MCP-1 / CCR2 signaling axis.