大麻素2型受体激活对钛颗粒诱导成骨细胞凋亡的干预作用  

Type-2 cannabinoid receptor agonist inhibits titanium particle-induced osteoblast apoptosis

作　　者：裴方[1] 周冰[1] 郑伟[1] 沈振宇[1] 王琛[1] 仇尚 郭开今[1] 

机构地区：[1]徐州医学院附属医院,江苏省徐州市221002

出　　处：《中国组织工程研究》2016年第43期6431-6437,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:成骨细胞凋亡而导致的成骨减少是最终假体松动的重要原因,大麻素2型受体的激活可促进成骨细胞的增殖,减少细胞凋亡率的发生。目的:探讨钛颗粒负荷下,大麻素2型受体激动剂HU-308对成骨细胞株MC3T3-E1凋亡的抑制作用及保护机制。方法:将体外培养的小鼠成骨细胞MC3T3-E1分5组培养,空白对照组以常规细胞培养基培养;钛颗粒组以含2.5 g/L钛颗粒的细胞培养基培养;低浓度药物组以含2.5 g/L钛颗粒+10^(-9) mol/L HU-308的细胞培养基培养;中浓度药物组以含2.5 g/L钛颗粒+10^(-8) mol/L HU-308的细胞培养基培养;高浓度药物组以含2.5 g/L钛颗粒+10^(-7)mol/L HU-308的细胞培养基培养。培养48 h,检测各组细胞增殖活性、凋亡、细胞内活性氧水平、线粒体膜电位水平及Caspase-3活性。结果与结论:(1)与空白对照组比较,钛颗粒组细胞增殖活性、线粒体膜电位水平明显下降(P<0.01或P<0.05),凋亡率、细胞内活性氧水平、Caspase-3活性明显升高(P<0.01或P<0.05);(2)与钛颗粒组比较,高浓度药物组细胞增殖活性、线粒体膜电位水平升高(P<0.05),中、高浓度药物组细胞凋亡率、细胞内活性氧水平、Caspase-3活性降低(P<0.01或P<0.05);(3)结果表明,HU-308能有效逆转钛颗粒对MC3T3-E1成骨细胞活性的抑制及促凋亡作用,而这些保护作用可能是HU-308通过调节细胞内活性氧水平、提升线粒体膜电位水平及Caspase-3活性来实现的。BACKGROUND： Impaired osteogenesis caused by osteoblast apoptosis is a main reason of aseptic loosening. Type-2 cannabinoid receptor activation can promote osteoblast proliferation and reduce apoptosis. OBJECTIVE; To investigate the inhibitory effect of HU-308 （type-2 cannabinoid receptor agonist） on the osteoblastic MC3T3-E1 cells co-cultured with titanium particles and its relative mechanism. METHODS： Mouse MC3T3-E1 cells were cultured in vitro and divided into five groups： cells were cultured in the normal medium as blank control group, cultured in the medium with 2.5 g/L titanium particles as titanium group, cultured in the medium containing 2.5 g/L titanium particles plus 10^-9, 10^-8 and 10^-7 mol/L HU-308 as low-, medium- and high-concentration groups, respectively. Forty-eight hours later, cell proliferation, apoptosis, levels of reactive oxygen species and mitochondrial membrane potential as well as Caspase-3 activity were assessed. RESULTS AND CONCLUSION： Compared with the black control group, the cell proliferation activity and the level of mitochondrial membrane potential were significantly reduced and the apoptosis rate, the level of reactive oxygen species and Caspase-3 activity were significantly increased in the titanium group （P 〈 0.01 or P 〈 0.05）. Compared with the titanium group, the cell proliferation activity and the level of mitochondrial membrane potential significantly increased in the high-concentration group, and the apoptosis rate and Caspase-3 activity significantly decreased in the medium- and high-concentration groups （P 〈 0.05）. These findings demonstrate that HU-308 can effectively improve the impaired osteogenesis induced by titanium particles, probably through modulating reactive oxygen species and elevating mitochondrial membrane potential and Caspase-3 activity.

关 键 词：钛 成骨细胞 细胞凋亡 组织工程 生物材料 骨生物材料 大麻素2型受体 钛颗粒 MC3T3-E1细胞 细胞内活性氧水平 线粒体膜电位 CASPASE-3活性 流式细胞 酶联免疫吸附实验 

分 类 号：R318[医药卫生—生物医学工程]