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乙醇对成骨细胞凋亡影响的体外实验研究
An Experimental Study of the Influence of Alcohol on Osteoblast Apoptosis in Vitro
【摘要】 目的探讨乙醇对成骨细胞凋亡的影响。方法体外培养新生大鼠成骨细胞作为细胞模型,对照组采用含10%FBS的1640培养液培养;干预组在对照组正常培养1d后,加入100mmol/L乙醇。取第3代成骨细胞按实验分组,于乙醇作用后1、3、5、7d进行形态学观察及成骨细胞增殖的检测,于乙醇作用后3、7、10、14d进行成骨细胞分化指标检测,于乙醇作用后1d采用流式细胞仪对成骨细胞凋亡进行检测。结果2组第1天细胞增殖差异无统计学意义(P>0.05),干预组第3、5、7天细胞增殖率低于对照组,差异有统计学意义(P<0.01)。培养后3、7、10、14d,对照组细胞碱性磷酸酶(ALP)、骨钙素(BGP)逐渐升高,干预组细胞ALP、BGP逐渐降低(P<0.05);同期比较,干预组细胞ALP、BGP含量低于对照组(P<0.01)。对照组成骨细胞1d自发性凋亡率低于干预组[(1.89±0.63)%vs(2.94±0.91)%],差异有统计学意义(P<0.01)。结论一定浓度的乙醇可以诱导体外培养成骨细胞发生凋亡,造成其增殖与分化能力逐渐减弱。
【Abstract】 Objective To study the influence of alcohol on osteoblast apoptosis in vitro. Methods The mouse calvarial osteoblasts were cultured in vitro to three generations and divided into two groups: the control group and intervention group. Cells of control group were cultured in 1640 medium containing 10% FBS, and cells of intervention group were added 100 mmol/L alcohol after 1 day normal culture. After 1 , 3 , 5 and 7d culture, cells were harvested to measure MTT value. After 3 , 7 , 10 and 14 d culture, cells were harvested to measure alkaline phosphatase (ALP) and osteocalcin (BGP) activity. After 1 d culture, the osteoblast apoptosis was assessed by flow cytometry. Results There was no significant difference in the value of MTT between two groups (P > 0.05). The cellular proliferation rate was significantly lower in intervention group after 3, 5 and 7 d culture (P < 0.01). The values of ALP and BGP significantly increased in control group and decreased in intervention group after 3, 7, 10 and 14 d culture (P < 0.05 or P < 0.01). The values of ALP and BGP were significantly lower in interven tion group than those in control group after 3, 7, 10 and 14 d culture (P < 0.01). The apoptotic rate was significantly higher in intervention group than that in control group after 1 d culture (P < 0.01). Conclusion Alcohol had a marked effect on osteoblast apoptosis by weakening the proliferation and differentiation.
【Key words】 ethanol; osteoblasts; apoptosis; cell proliferation; alkaline phosphatase; osteocalcin;
- 【文献出处】 天津医药 ,Tianjin Medical Journal , 编辑部邮箱 ,2013年02期
- 【分类号】R965
- 【被引频次】2
- 【下载频次】91