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目的探讨在微应变环境下,柚皮苷体外对培养的骨髓间充质干细胞(MSCs)向成骨细胞分化的影响及其机制。方法将体外培养的兔MSCs接种于硅橡胶膜上,利用EF3200力学试验仪搭载的BioDynamic生物反应舱系统,对细胞进行周期性循环牵张微应变(微应变)加载。实验分成8组:对照组(常规培养),微应变组,柚皮苷不同质量浓度(2、20、200 ng/mL)组,微应变+柚皮苷(2、20、200 ng/mL)组。流式细胞仪检测各组MSCs的增殖指数;RT-PCR检测各组MSCs骨钙蛋白(OCN)、成骨特异性转录因子(Runx 2)与I型胶原(Col I)基因的表达。结果在微应变环境下,柚皮苷使MSCs呈现明显的极性排列趋向,且细胞长轴平行力学刺激方向;显著提高MSCs的增殖活性;柚皮苷200 ng/mL显著上调OCN基因表达;不同质量浓度的柚皮苷均显著上调Runx 2基因表达,且与质量浓度呈正相关;低质量浓度柚皮苷促进Col I基因表达,而高质量浓度柚皮苷抑制了Col I基因的表达。结论柚皮苷可增强在微应变环境下MSCs的增殖并能够促进其向成骨分化。
Objective To investigate the effect of naringin on the differentiation of cultured bone marrow mesenchymal stem cells (MSCs) into osteoblasts under microstrain and its mechanism. Methods Rabbit MSCs cultured in vitro were inoculated on a silicone rubber membrane and the cells were loaded with cyclic strain microdissection (strain) by BioDynamic biological reactor system equipped with EF3200 mechanical tester. The experiment was divided into 8 groups: the control group (routine culture), the micro-strain group, the naringin group with different concentrations (2,20,200 ng / mL), the micro-strain + naringin group. The proliferation index of MSCs in each group was detected by flow cytometry. The expression of osteocalcin (OCN), osteoblast-specific transcription factor (Runx 2) and type I collagen (Col I) were detected by RT-PCR. Results Under the condition of micro-strain, naringin marked the orientation of MSCs, and the long axis of the cells acted parallel to the direction of mechanical stimulation. The proliferation activity of MSCs was significantly increased. Naringin 200 ng / mL significantly upregulated the expression of OCN gene. Mass concentration of naringin significantly up-regulated Runx 2 gene expression, and was positively correlated with mass concentration; low-concentration naringin promoted Col I gene expression, while high-concentration naringin inhibited Col I gene expression. Conclusion Naringin can enhance the proliferation of MSCs under micro-strain environment and promote its differentiation into osteoblasts.