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乙对成骨细胞凋亡影响的体外实验研究

肖瑜1,张凯2,张福江2,陆芸1   

  1. 1. 天津市天津医院
    2. 天津医院
  • 收稿日期:2012-02-08 修回日期:2012-06-21 出版日期:2013-02-15 发布日期:2013-02-15
  • 通讯作者: 肖瑜

The influence of alcohol on osteoblast apoptosis in vitro

  • Received:2012-02-08 Revised:2012-06-21 Published:2013-02-15 Online:2013-02-15
  • Contact: XIAO Yu

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摘要：

【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1 d后, 加入100 mmol/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、7 d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、14 d进行成骨细胞分化指标检测，于乙醇作用后1 d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、14 d，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1 d 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。

关键词: 乙醇, 成骨细胞, 凋亡

Abstract:

[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 orP < 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 osteo? blast apoptosis by weakening the proliferation and differentiation.

Key words: alcohol osteoblast apoptosis