| Objective:By detecting the expression of CFTR,Wnt/?-catenin signaling pathway(Wisp-1,Axin and ?-catenin)in tooth germs of coal-burning dental fluorosis rat offsprings,to investigate the expression of CFTR and Wnt/?-catenin signaling pathway during enamel development.To determine the key target time that fluorine effect on CFTR,simultaneously,preliminarily discussing the underlying mechanism of dental fluorosis.This study will provide a new theoretical basis for specifically blocking the incidence of dental fluorosis.Methods: 1.Our previous experiment had successfully copied the coal-burning dental fluorosis rat model,based on it,the study continued to produce offspring.Twelve tooth germ tissues were randomly gathered after postnatal day 0(P0),P1,P7,P14 and P 20,which were in the enamel developmental stages.2.Real-time fluorescence quantitative PCR was used to detect the expression of CFTR m RNA in tooth germs in above time points.3.Western blot and Real-time fluorescence quantitative PCR were utilized to detect the level of ?-catenin protein,the expression of Wisp-1 and Axin gene in tooth germs in different points.Results:1.In the control group,the relative expression of CFTR was gradually up-regulated with time,it increased significantly in the P14 and P20,and there were statistical differences compared to the expression levels of P0,P1,and P7(P?0.01).In experimental group,CFTR m RNA expression showed a downward trend with the increase of the time.In the P14 and P20 it decreased more than P0,P1,and P7(P?0.01).Compared with the control group,the expression of CFTR in the experimental group of P0 and P1 offspring tissues was significantly higher than that of the control group(P?0.01).In P7,P14 and P20,the expression were lower than the control group,the difference was statistically between significant in P14 and P20(P?0.01).2.The level of ?-catenin protein in tooth germs of both control and experimental groups increased with time,and the increase in experimental group was more significant.There was a statistically significant difference between the two groups at each of the adjacent time points in the same group(P<0.05),except for P0 and P1.The expression levels of the experimental groups at different time points were higher than those of the control group,and the difference was statistically significant(P<0.05).3.The expression of Wisp-1 m RNA increased with the increase of time in the experimental group and the control group.The increase of Wisp-1 m RNA in the experimental group was more significant,and there was statistically significant difference between two groups(P<0.05),except the P0.4.Axin m RNA expression was gradually increased from P0 to P14 in normal tooth germs tissues,while Axin expression was down-regulated in P20.In the experimental group,Axin m RNA expression decreased in P1,increased in P7,decreased again in P14,and continued to increase in P20.we observed that the expression of Axin in experimental group was higher than the control group in P0 and P20,and the difference was statistically significant(P?0.05).In P7,P14 and P20,the expression levels were lower than those in the control group,there was statistically significant differences between two groups(P<0.05),except for P7.Conclusion: 1.Excessive fluorine can affect the expression of CFTR in ameloblast maturation(enamel mineralization).2.In the high fluoride environment,inhibition of CFTR expression may activate Wnt/?-catenin during the maturation period of ameloblast.3.In ameloblast maturation,CFTR down-regulation and Wnt/?-catenin signal pathway over-activation may be related to the pathogenesis of coal-burning dental fluorosis. |
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