王长松,桂传枝,刘家骝,于燕妮,唐俊杰,陈燕平
WANG Chang-song 1; GUI Chuan-zhi 1; LIU Jia-liu 1; YU Yan-ni 1; TANG Jun-jie 2; CHEN Yan-ping 3 (1.Department of Pathology;

• 1:贵阳医学院病理学教研室
• 2:贵阳医学院病理学教研室
• 3:贵阳医学院病理学教研室
• 4:贵阳医学院病理学教研室

摘要(Abstract)：

目的 :建立成骨细胞体外培养模型 ,并观察不同浓度的氟对成骨细胞的影响 ,为氟中毒的机制研究提供实验依据。方法 :用幼兔肋骨培养成骨细胞 ,并纯化、鉴定。用不同浓度的氟化钠处理 (2 0 ,16 0 ,2 4 0 ,4 0 0μmol/L)成骨细胞 ,MTT法测定氟对成骨细胞增殖活性的影响 ;流式细胞术测定细胞周期变化及凋亡情况。 结果 :低浓度的氟化钠 (2 0 μmol/L)在体外可显著促进成骨细胞增殖 (P <0 0 1) ,不引起成骨细胞的凋亡 ,可使S期和G2 /M期的细胞明显增多 ;高浓度的氟化钠 (16 0 ,2 4 0 ,4 0 0 μmol/L)则抑制成骨细胞的增殖 (P <0 0 1) ,引起成骨细胞的凋亡 (P <0 0 1和P <0 0 5 ) ,G2 /M期细胞明显减少。结论 :(1)成功建立了氟中毒的体外细胞模型 ;(2 )低浓度的氟化钠可以促进成骨细胞的增殖。高浓度的氟化钠抑制成骨细胞的增殖 ,诱导细胞的凋亡 ,抑制细胞由S期向G2 /M期转化。氟对成骨细胞的作用具有双向性
Objective: To culture the osteoblasts and study the influence of fluoride on osteoblasts in vitro. Methods: Osteoblasts were cultured, purified and appraised, then dealt with various concentrations of fluoride (20, 160, 240 and 400 μmol/L). The proliferation of osteoblasts was observed with MTT method, and the changes of cell phase and apoptosis were investigated using flow cytometry. Results: Fluoride of low concentration (20 μmol/L) promoted the proliferation of osteoblasts in vitro (P<0.01) and the cells in S and G2/M phase increased markedly. It did not increase the apoptosis of osteoblasts. With the increasing concentrations of fluoride (160,240 and 400 μmol/L), the proliferation of osteoblasts was inhibited. The apoptosis was induced and the cell transformation from S phase to G2/M phase was also inhibited. Conclusions: 1) The model of cell fluorosis has been established in vitro successfully; 2) Fluoride of low concentration could promote the proliferation of osteoblasts and increase the cell number in S and G2/M phases; Fluoride of high concentrations could inhibite the proliferation of osteoblasts and induce the apoptosis; 3) Fluoride has two-phase effects in vitro.

关键词(KeyWords)： 氟;成骨细胞;细胞周期;细胞凋亡;兔
fluorine; osteoblasts; cell cycle; apoptosis; rabbits

Abstract:

Keywords:

基金项目(Foundation): 贵州省教育厅资助项目 (F99 3)

作者(Authors): 王长松,桂传枝,刘家骝,于燕妮,唐俊杰,陈燕平
WANG Chang-song 1; GUI Chuan-zhi 1; LIU Jia-liu 1; YU Yan-ni 1; TANG Jun-jie 2; CHEN Yan-ping 3 (1.Department of Pathology;

DOI: 10.19367/j.cnki.1000-2707.2004.03.005

参考文献(References)：

• [1]王前,钟世镇,龚文汇,等.鼠颅盖骨成骨细胞体外培养及ALP免疫组化法鉴别纯化[J].中华骨科杂志,1995,15(6):364-366.
• [2]陈奇.中药药理研究方法学[M].北京:人民卫生出版社,1994.834.
• [3]崔晓暄,王士雯,齐鹏.主动脉中膜细胞体外钙化的研究[J].中华老年医学杂志,2000,19(6):447-449.
• [4]FarleyJR ,WergedalJE ,BarlinkDJ.Fluoridedirectlystimulatesproliferationandalkalinephosphateseactivityofbone formingcells[J].Science,1983,222(4621):330-332.
• [5]KassemM ,MosekildeL ,EriksenEF .1,25DihydroxyvitaminD3potentiatesfluoride stimulatedcollagentypeIproductionincul turesonhumanbonemarrowstromalosteoblast likecells[J].JBoneMinerRes,1993,8(12):1453-1458.
• [6]ReedBY ,ZerwekhJE ,AntichPP ,etal.Fluoridestimulated
• [3H]thymidineuptakeinahumanosteoblasticosteosarcomacelllineisdependentontransforminggrowthfactor[beta][J].JBoneMinerRes,1993,8(1):19-25.
• [7]BurgenerD ,BonjourJP ,CaverzasioJ.Fluorideincreasestyro sinekinaseactivityinosteoblast likecells:RegulatoryroleforthestimulationofcellproliferationandPitransportacrosstheplasmamembrane[J].JBoneMinerRes,1995,10(1):164-171.
• [8]KassemM ,MosekildeL ,EriksenEF .Effectsoffluorideonhu manbonecellsinvitro:Differencesinresponsivenessbetweenstromalosteoblastprecursorsandmatureosteoblasts[J].ActaEndocrinol,1994,130(1):381-386.
• [9]井玲,邵宗俊,任立群,等.氟中毒大鼠肝细胞凋亡研究[J].中国地方病学杂志,1999,18(2):84-86.

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