| Objective: Altered the binding of bone morphology and genetics aspects, to explore fluorine and aluminum toxicity in rats spine and possible pathogenesis. Methods: We divided 96 rats into 6 groups, respectively as control group, mid-fluoride group, high-fluoride group, mid-fluoride-aluminum group, high-fluoride-aluminum group and pure-aluminum group. Each group includes 16 rats,half male and half female. Feed each group of rats with forage contains Na F and aluminum-contaminated water which contains Al Cl3 continuously for 6 months, the concentration of each group are respectively(0mg/kg,0mg/L)、(45mg/kg,0mg/L)、(110mg/kg,0mg/L) 、(45mg/kg,90mg/L) 、(110mg/kg,90mg/L) 、(0mg/kg,90mg/L). Oberseve the rats’ common growing condition and collect 24 h urine before executing rats. After the execution, preserve the spines and thigh-bones. Measure the urinary fluoride level and skeletal fluoride level with F-ion selective electrode method, and the urinary aluminum level and skeletal aluminum level with Inductively Coupled Plasma-Atomic Emission Spectrometry method. Observe the change of the trabecular bones of the rats with microscopic CT, and protein expression level of the rats spine with ELISA method. Extract the total RNA of rats spine tissues, and establish a real-time orescent quantitative RT-PCR method for detection of the expression level of Runx2 and Osx m RNA. Results: 1.The development of Fluoride-Aluminum poisoned animals model:no significant occurrence of dental fluorosis in control group and Aluminum group, while in other groups the incidence rate and severity of dental fluorosis increases with the prolonged exposure time. In comparison, the urinary fluoride and skeletal-fluoride levels of the rats are relatively higher in the Fluoride-groups than those in the control group, the difference was statistically significant(P<0.05). Combining the dental fluorosis conditions and indicators such as urinary fluoride,skeletal-fluoride, urinary aluminum and skeletal aluminum level, it is believed that this research has developed Fluoride-Aluminum poisoned animals model.2.Observe the change of the rats trabecular bones with microscopic CT: compare the change of BMD, TMD, Tb.Th, Tb.Sp, Tb.N among each groups.The difference was statistically significant(P<0.05). Compared with the control group, the DA, BS/BV of fluoride-groups decreased while the BV/TV are higher. But comparing the changes of DA, BS/BV and BV/TV among each group, the difference was not statistically significant(P>0.05). Analyze the effect fluoride and aluminum have on trabecular bones with analysis of variance of factorial design, no interaction between the two factors were found. 3. Compared with the control group, the expression level of Runx2 and Osx m RNA in each fluoride-groups are higher, and the difference was statistically significant.(P<0.05), while the expression levels in the pure-aluminum group are relatively lower and the difference was not statistically significant(P>0.05). The interaction between fluoride and aluminum,and the main effect fluoride and aluminum have on Runx2 and Osx m RNA were both not statistically significant(P>0.05).Compared with the control group, the content of Runx2 protein are higher in the poisoned groups. Compared with the pure-fluoride group, the con-tent of Runx2 protein in the fluoride-aluminum group and control group increases along the fluoride level. The differences were both statistically significant(P<0.05). Except the mid-fluoride group,the Osx protein level are lower in the control group than in each poisoned group(P<0.05). The analysis of the interaction between fluoride and aluminum indicates that the main effect fluoride and aluminum have, and the interaction between fluoride and aluminum are both statistically significant(P<0.05). Conclusion: 1.Developed the Fluoride-Aluminum poisoned animals model successfully.2.Fluoride can promote rats accelerated bone turnover, increase bone mass, pure fluoride group in rats bone sclerosis mainly skeletal fluorosis; Alcan alone occur in rats osteomalacia mainly skeletal fluorosis; aluminum and fluorine poisoning rats appeared osteopetrosis and osteomalacia hybrid skeletal fluorosis performance.3. The interaction between fluoride and aluminum of Runx2 and Osterix gene protein in rats spine and fluoride content of urinate and bone present to be antagonism. |
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