摘要
目的观察氟对体外培养乳兔肋软骨的直接作用,探讨氟对软骨细胞形态和细胞凋亡的影响。方法采用乳兔的肋软骨体外培养方法,实验分为处理组和对照组。处理组根据加氟浓度的高低(20μmol/L、320μmol/L),加或不加SOD(0.006U/L)分为4组。培养18d后观察软骨细胞柱的形态学变化并做计量学分析;观察细胞超微结构变化;用TUNEL法检测细胞凋亡并计算凋亡指数(AI,%);测定细胞生长液中H2O2和NO水平。结果①光镜下,高氟组软骨增生区的软骨细胞数量增多,细胞柱较对照组明显增长,但变细,其余各处理组细胞柱也变细长。与对照组相比,2个单纯加氟组软骨细胞柱长度、面积和周长增加,细胞柱横径减少;2个氟+SOD组软骨细胞柱的改变与2个单纯加氟组相比程度明显减轻。电镜下,4组均有凋亡的特征性改变,以单纯高氟组为最多。②与对照组相比,高氟组细胞凋亡指数(AI)明显增高(P<0.01);与单纯加氟组相比,加SOD组细胞AI值明显下降(P<0.05,P<0.01)。③H2O2水平在2个高氟组与对照组比较,差异有显著意义(P<0.05,P<0.01)。除低氟+SOD组外,其余各组之NO浓度均升高,和对照组相比,差异有显著意义(P<0.01,P<0.05)。结论①氟可引起发育过程中的软骨细胞柱形态改变,损害细胞的超微结构。②氟可引起乳兔肋软骨中细胞凋亡增加。
Objective To observe the direct influence of fluoride on rib cartilage of new-born rabbit in vitro, then discussing the effect of fluoride on morphologic changes of cell columns and apoptosis of cartilage cells. Methods The coastal cartilages of newborn-rabbits were cultured with fluoride in various concentration (20?320 μmol/L) with/without SOD(0.006 U/L) for 18days. Then examined: morphologic changes with image analysis and ultra-structural changes;apoptosis was detected with terminal deoxyribonucleotide transferase-mediated dUTP-bintin(TUNEL) method; and the concentration of H2O2 and NO in the culture medium were tested. Results ①High concentration fluoride treated the chondrocytes were proliferated, cell columns being longer but thinner when being compared with the control, the perimeter, area and length of cartilage columns increased, but the cross diameter decreased in the two fluoride treated groups. The above-mentioned changes were markedly alleviated in the two fluoride plus SOD groups. It was observed the characteristics of apoptosis in every fluoride-treated groups, but it was serious in 320 μmol/L group than other groups. ②The AI (%) of fluoride-treated groups (320 μmol/L) was remarkably high as being compared with the control (P < 0.01),the AI value of SOD plus fluoride decreased significantly when compared with fluoride-treated groups (P < 0.05,P < 0.01). ③The H2O2 content in the two groups-treated with high concentration fluoride increased markedly when compared with the control (P < 0.05, P < 0.01);the NO content in treated groups was markedly higher than that of the control(P < 0.01, P < 0.05) except for low concentration fluoride plus SOD group. Conclusions ①Fluoride causes the changes of cell columns of cartilage and damages the cell ultrastructure. ②In vitro, fluoride induced apoptosis of new-born rabbit cartilage chondrocytes and SOD can inhibit it. ③Excessive free radicals partly responsible for the mechanisms of fluoride induced apoptosis.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2004年第5期423-426,共4页
Chinese Jouranl of Endemiology
基金
贵州省教委科学基金资助项目(F99-3)
