绿原酸对缺氧环境下干细胞来源软骨样细胞凋亡的影响

Protective effect of chlorogenic acid on differentiated chondrogenic MSCs apoptosis induced by hypoxia

目的探讨缺氧环境下绿原酸对骨髓间充质细胞来源软骨样细胞(Chondrogenic MSCs)影响及机制。方法采用体外培养大鼠间充质干细胞,以含0.2 mg.L-1 BMP-2诱导液培养12 d使其成为软骨样细胞后进行实验。分为(A)正常对照组、(B)绿原酸组、(C)0.1%O2缺氧环境组、(D)绿原酸+0.1%O2缺氧环境组。检测各组Hoechst33258荧光染色检测细胞凋亡、细胞内活性氧水平;RT-PCR检测不同组软骨细胞Caspase-3和Bcl-2基因的表达。结果与正常对照组比较,0.1%O2缺氧环境12 h能明显造成干细胞来源软骨样细胞的凋亡(P<0.05),绿原酸可降低0.1%O2缺氧引起的软骨样细胞凋亡率(P<0.05),活性氧水平下降,Bcl-2表达增强,Caspase-3表达减弱。结论绿原酸可抑制0.1%O2缺氧引起的软骨样细胞凋亡,其作用机制可能与降低细胞内的活性氧水平,稳定细胞的氧化还原状态来保护线粒体膜电位,促进凋亡抑制基因Bcl-2的表达及抑制Caspase-3的表达有关。

Aim To investigate the effect of chlorogenic acid on the injury of chondrocytes differentiated from rat marrow stromal cells induced by hypoxia in vitro and to explore its underlying mechanism.Methods The chondrocytes differentiated from rat marrow stromal cells（5 weeks rats） by 0.2 mg·L-1 BMP-2 for 12 days.This study was conducted as follows：（A）normal control group,（B）chlorogenic acid group,（C）hypoxia injury group,（D）chlorogenic acid and hypoxia injury group.Cell apoptosis was observed by Hoechst33258;reactive oxygen species was also detected;chondrocytes Bcl-2 gene and Caspase-3 changes were detected by RT-PCR.Results Compared with normal control group,hypoxia induced damage of differentiated chondrogenic MSCs（P0.05）.Chlorogenic acid could disturb the damage of differentiated chondrogenic MSCs induced by hypoxia.It showed a reduction in the rate of apoptosis（P0.05）.Reactive oxygen species was decreased,the gene expression of Caspase-3 was inhibited,and the gene expression of Bcl-2 was increased.Conclusion Chlorogenic acid can inhibit the apoptosis of chondrocytes differentiated from rat marrow stromal cells induced by hypoxia,and its mechanism may be related to protecting mitochondrial membrance potential,increaseing the gene expression of Bcl-2 and inhibiting the gene expression of Caspase-3.