Objective To study the expression of silent information regulator (SIRT) in brains of rats with chronic fluorosis and reveal the correlation between SIRT1 and the ability of learning and memory of rats.Methods Sixty SD rats were selected and their body weight was (100 ± 20) g,according to the body mass of the rats,random number table method was used to divide rats into control group,low and high fluoride groups,experimental period was 3 and 6 months (ten rats in each experimental period,half males and half females).In control group,the rats were fed with drinking water containing no more than 0.5 mg/L fluoride;the rats in low and high fluoride groups were fed drinking water containing 5.0 and 50.0 mg/L fluoride,respectively.All of rats were fed the same standard food containing no more than 0.6 mg/kg fluoride.Three degree method was used to check the formation of dental fluorosis.Rat urinary fluoride was determined via the fluoride electrode method;Morris water maze method was used to detect the ability of learning and memory of rats (the escape latency time,the number of crossing the platform and stay time in platform quadrant);the protein and mRNA expression levels of SIRT1 were detected by Western blotting and Real-time PCR,respectively.Results In the experimental period of 3 and 6 months,no dental fluorosis was observed of rats in control group,but there were different degrees of dental fluorosis in low and high fluoride groups,especially in high fluoride group.The urinary fluorine contents [(1.60 ± 0.09),(1.91 ± 0.16) mg/L;(1.94 ± 0.19),(2.31 ± 0.18) mg/L] of rats fed with low and high fluoride for 3 or 6 months were significantly higher than those in control group [(1.08 ± 0.15),(1.09 ± 0.17) mg/L,P ＜ 0.05].The escape latency time [(18.36 ± 2.80) s] of rats in the high fluoride group at the end of 3 months was higher than that of control group [(6.68 ± 3.01) s,P ＜ 0.05],the number of stay time in platform quadrant [(12.91 ± 3.25) s] was lower than that of control group [(19.97 ± 3.30) s,P ＜ 0.05].The escape latency time [(15.46 ± 4.56),(28.16 ± 4.00) s] of rats in low and high fluoride groups at the end of 6 months were all higher than that of control group [(6.62 ± 2.31) s,P ＜ 0.05];the number of crossing the platform and stay time in platform quadrant [(2.25 ± 1.71) times,(12.73 ± 3.55) s;(1.40 ± 1.15) times,(9.26 ± 1.72) s] of these rats were significantly lower than those of the control group [(4.00 ± 1.58) times,(19.53 ± 4.36) s,P ＜ 0.05].The expression levels of protein [(73.84 ± 9.68)％,(73.23 ± 4.51)％;(53.30 ± 17.63)％,(54.69 ± 18.71)％] and mRNA [(70.33 ± 4.89)％,(66.27 ± 3.38)％;(37.72 ± 4.89)％,(44.15 ± 1.74)％] of SIRT1 in the hippocampus and cortex of rats fed with high fluoride for 3 or 6 months were significantly lower than those in control group [(100.00 ± 13.51)％,(100.00 ± 13.60)％;(100.00 ± 15.37)％,(100.00 ± 12.19)％;(100.00 ± 2.65)％,(100.00 ± 4.34)％;(100.00 ± 3.40)％,(100.00 ± 4.52)％,P ＜ 0.05].Whereas,the decreased expression levels of protein [(77.65 ± 14.51)％,(71.51 ± 8.27)％] and mRNA [(57.78 ± 1.96)％,(63.76 ± 2.16)％] of SIRT1 in the hippocampus and cortex of rats in the low fluoride group were only observed at the end of 6 month of experiment (P ＜ 0.05).The expression of protein of SIRT1 in the hippocampus and cortex of rats in 3 or 6 months was negatively correlated with the escape latency time of rats (r=-0.598 5,-0.493 2;-0.782 6,-0.777 3,P＜ 0.05),and it was positively correlated with the number of crossing the platform (r =0.547 7,0.523 3;0.720 5,0.715 4,P ＜ 0.05).Conclusion The decrease of the ability of learning and memory in rats with chronic fluorosis may be related to the decreased expression of SIRT1 influenced by chronic fluorosis.

Objective To detect the expression of peroxisome proliferator-activated receptor γ (PPARγ) in the brains of rats with chronic fluorosis and elucidate the relationship between PPARγand oxidative stress in chronic fluorosis.Methods According to body weight (100-120 g),sixty healthy SD rats were divided into control group (less than 0.5 mg/L fluoride in drinking water),low fluoride group (5.0 mg/L fluoride in drinking water,prepared by NaF),and high fluoride group (50.0 mg/L fluoride in drinking water) via the random number table method,20 rats in each group (half male and half female).The experiment periods were 3 and 6 months,respectively.Then 24-hour urine samples of rats were collected from each group,all rats were put to death and brain tissues were taken.The fluoride contents in urine and brain tissue were measured with fluoride-ion selective electrode;the levels of PPARγ protein and mRNA in the cortex and hippocampus were determined by Western blotting and Real-time fluorescence quantitative PCR,respectively;and the activities of superoxide dismutase (SOD) and level of malondialdehyde (MDA) in serum were detected by xanthine oxidase method and thiobarbituric acid method;the correlation between PPARγ protein expression and oxidative stress was analyzed.Results After 3 and 6 months of treatment,the contents of fluoride in urine and brain in low fluoride group [(1.57 ± 0.18) mg/L,(3.43 ± 0.70) μg/g;(1.79 ± 0.17) mg/L,(7.40 ± 1.21) μg/g] were higher than those of control group [(1.11 ± 0.17) mg/L,(2.39 ± 0.50) μg/g;(1.02 ± 0.15) mg/L,(2.87 ± 0.82) μg/g,P ＜ 0.05],and the values in high fluoride group [(1.91 ± 0.23) mg/L,(6.70 ± 0.87) μg/g;(2.44 ± 0.51) mg/L,(12.10 ± 1.30) μg/g] were significantly higher than those in low fluoride group (P ＜ 0.05).In high fluoride group after 3 months of treatment,the expression of PPARγprotein [(79.00 ± 3.46)％,(80.35 ± 2.50)％] and mRNA [(79.11 ± 11.18)％,(82.10 ± 9.94)％] in hippocampus and cortex of rat brains were significantly lower than those of low fluoride group [(104.01 ± 5.77)％,(101.17 ± 6.35)％;(112.88 ± 22.15)％,(101.14 ± 8.60)％,P＜ 0.05];the expression of PPARγprotein [(64.32 ± 10.43)％,(60.20 ± 10.92)％] and mRNA [(41.03 ± 9.93)％,(52.25 ± 11.48)％] in the same brain regions of the rats after 6 months of treatment in high fluoride group were significantly lower than those of control group [(99.99 ± 11.19)％,(100.00 ± 11.30)％;(100.00 ± 10.00)％,(100.00 ± 9.00)％] and low fluoride group [(73.88 ± 3.36)％,(81.50 ± 14.90)％;(76.02 ± 8.65)％,(73.36 ± 7.43)％,P ＜ 0.05].The activities of SOD in serum in low and high fluoride groups after 6 month treatment [(37.94 ± 1.92),(35.54 ± 2.53) U/ml] were significantly lower than that of control group [(41.24 ± 0.66) U/ml,P ＜ 0.05],and the value in high fluoride group was lower than that in low fluoride group (P ＜ 0.05);serum MDA contents in high fluoride group after 3 and 6 month treatment [(8.29 ± 1.49),(11.63 ± 1.04) nmol/mg pr] were higher than those in low fluoride group [(6.39 ± 0.69),(7.50 ± 1.64) nmol/mg pr] and control group [(5.02 ± 0.71),(5.87 ± 1.03) nmol/mg pr,P ＜ 0.05].The correlation analysis results showed the levels of PPARγprotein in hippocampus and cortex of rats were negatively correlated with fluoride contents in brain tissues (3 month:r=-0.769,-0.793;6 month:r =-0.832,-0.870;P ＜ 0.05),positively correlated with SOD activities (3 month:r =0.550,0.826;6 month:r =0.822,0.896;P ＜ 0.05) and negatively correlated with MDA contents (3 month:r =-0.703,-0.609,6 month:r =-0.792,-0.657;P ＜ 0.05) in serum.Conclusions Declined expression of PPARγat protein and mRNA levels has been detected in brains of rats with chronic fluorosis,which might be related to the increase of oxidative stress.PPARγ may be involved in the occurrence of chronic fluorosis.