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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,抗氧化剂SOD可以抑制
Objective To observe the direct effect of fluorine on cultured rabbit cartilage costal cartilage in vitro and to explore the effect of fluorine on chondrocyte morphology and apoptosis. Methods Rabbit costal cartilage cultured in vitro methods, the experiment divided into treatment group and control group. The treatment group was divided into 4 groups according to the fluoride concentration (20μmol / L, 320μmol / L), with or without SOD (0.006U / L). The morphological changes of chondrocytes were observed after 18 days of culture. The changes of ultrastructure of the chondrocytes were observed. The apoptosis of the chondrocytes was detected by TUNEL method and the apoptosis index (AI,%) was calculated. The contents of H2O2 and NO Level. Results ① Under the light microscope, the number of chondrocytes in cartilage hyperplasia area of high fluorine group increased, the number of cell column increased obviously compared with that of control group, but the number of cells became thinner, and the remaining cells of each treatment group also became slender. Compared with the control group, the length, area and perimeter of chondrocytes in two fluoride-only groups were increased, and the diameter of the column was decreased. The changes of chondrocytes in the two fluoride + SOD groups were similar to those in the two fluoride-only groups Obviously relieved. Under electron microscope, all the four groups had characteristic changes of apoptosis, with the highest in the group of high fluoride alone. Compared with the control group, the AI of the high fluoride group was significantly increased (P <0.01); compared with the fluoride-only group, the AI of the cells in the SOD group was significantly decreased (P <0.05, P <0.01) . (3) The level of H2O2 was significantly different between the two high fluoride groups and the control group (P <0.05, P <0.01). In addition to the low fluoride + SOD group, the other groups NO concentrations were increased, compared with the control group, the difference was significant (P <0.01, P <0.05). Conclusion ① Fluoride can change the morphology of chondrocyte during development and impair the ultrastructure of cells. ② fluoride can cause milk rabbit costal cartilage increased apoptosis, antioxidant SOD can be inhibited