摘要
目的 观察氟对体外培养软骨的成骨作用 ,进一步研究氟骨症发生的机理。方法 采用 10 d龄鸡胚股骨体外培养 ,培养基中加不同浓度的氟化钠 (12 0 ,2 40 ,480 ,960μmol/ L ) ,连续培养 11d。分别检测 :形态学及形态计量学指标 ;用 TUNEL法检测细胞凋亡 ,并计算细胞凋亡指数 (AI,% ) ;透射电镜观察鸡胚股骨细胞的超微结构 ;测定培养基中的 H2 O2 、NO及 SOD含量。结果 1加氟后软骨细胞体积变大 ,钙化增多。各加氟组的钙化面积、平均光密度、积分光密度和正常组相比均增多。 2细胞凋亡原位检测 :各加氟组的 AI和正常组相比显著增高 ,差异有显著性 (P <0 .0 5)。以 480μmol/ L组最高。 3超微结构变化 :各加氟组均出现凋亡细胞 ,随着氟浓度增加凋亡细胞数也增加 ,并有凋亡小体出现。 4培养基中 H2 O2 浓度在 480 μmol/ L 组开始升高 ,和正常组相比差异有显著意义 (P <0 .0 5) ;各加氟组的 NO浓度明显增加 ,和正常组相比差异有非常显著意义 (P <0 .0 1) ;SOD浓度从 2 40μm ol/ L F- 组开始下降 ,和正常组相比差异有显著意义 (P <0 .0 5)。结论 氟促进了软骨细胞的凋亡 ,使钙化增加 ,激发软骨内成骨过程 ,可能是氟骨症骨质增生型病变的发生机制 ,这和自由基浓度增高。
Objective To observe the effective of fluorid e on the bone formation in vitro and to explore the mechanisms of fluorosis.Methods The femur of chicken was cultured with fluorid e in various concentrations(120,240,480,960 μmol/L)for 11 days, then morpholo gic changes and calcification of cells were examined; apoptosis was detected by terminal deoxyribonucleotide transferase-mediated dUTP-bintin(TUNEL) method, a nd the apoptotic index(AI,%) was calculated; the ultrastructural changes of chon drocytes were observed with electron-microscope; at the same time the contents of H 2O 2,NO and SOD in the culture were tested.Results The mean values of calcification area, general light dense and sum light dense of fluoride-treated groups were higher than tho se of the control( P<0.05). The AI(%) of fluoride-treated groups was remark ably different as compared with the control (P<0.05 and P<0.01). It was observed there were apoptotic cells or apoptotic bodies in cells with electron -microscope. The H 2O 2 content increased in media of groups-treated with fl uoride above 240 μmol/L. (the differences were significant when compared with t he fluoride free control);the NO content in all fluoride-treated groups was mar kedly higher than that of the control group (P<0.01); the SOD activity was lower in all fluoride- treated groups (P<0.05) except for 120 μmol/L tha n that of control.Conclusions In vitro, fluoride promote apoptosis,calci fication and ossification. This is probably the mechanism by which osteofluorosi s occurred. Excessive free radicals might be responsible for the abnormal occurr ence of apoptosis, hence the excessive bone formation in case of fluorosis.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2003年第4期298-301,共4页
Chinese Jouranl of Endemiology
基金
贵州省教委科研项目 ( F 99- 3)
