Effects of different doses of sodium fluoride on cartilage lesion and expression of interleukin-6 in Balb/c mice / 中华地方病学杂志

ABSTRACT

Objective To explore the effects of different doses of sodium fluoride (NaF) on cartilage lesion and expression of interleukin-6 (IL-6) in serum and cartilage tissue of Balb/c mice.Methods Sixty-four 5-week-old male Balb/c mice were divided into 4 groups based on body weight via the random number table method and 16 mice were in each group.The mice in control group were fed with distilled water,and experimental animals in low,middle and high fluoride groups were fed with distilled water containing NaF 25,50 and 100 mg/L,respectively.The mice were weighed once a week and fed for three months to establish the drinking water fluorosis model.The fluoride contents in spine were detected via the fluorin-ion selective electrode method.The pathological changes in articular cartilage and epiphyseal plate cartilage were observed through optical microscope.The levels of serum IL-6 and souble IL-6 receptor (sIL-6R) were detected via the enzyme-linked immunosorbent assay.The expression of IL-6 protein in articular cartilage and epiphyseal plate cartilage was examined by immunohistochemistry.Results From the sixth week of the experiment,compared with other 3 groups,the body weight of high fluoride group decreased significantly (all P ＜ 0.05);from the seventh week,compared with control and low fluoride groups,the body weight of middle fluoride group decreased significantly (all P ＜ 0.05);throughout the experiment,compared with control group,the body weight of low fluoride group had not changed significantly (all P ＞ 0.05).The fluoride contents of bone in control group,low fluoride group,middle fluoride group and high fluoride group were (842.46 ± 89.27),(1 705.05 ± 105.76),(2 614.17 ± 156.10) and (3 444.58 ± 233.69) mg/kg,respectively.The differences between groups were statistically significant (F =309.716,P ＜ 0.05),and fluoride contents of bone increased with increase of fluoride doses (all P ＜ 0.05).Under optical microscope,the cartilage tissue of control group was normal,while articular cartilage and epiphyseal plate cartilage showed different degrees of cartilage ossification in fluorosis mice and the changes increased with the increase of fluoride doses.The levels of serum IL-6 in control group,low fluoride group,middle fluoride group and high fluoride group were (5.98 ± 1.43),(7.54 ± 2.16),(5.25 ± 1.97) and (6.31 ±-1.36) ng/L,respectively.The differences between groups were statistically significant (F =3.840,P ＜ 0.05),low fluoride group was significantly higher than control group (P ＜ 0.05),and middle fluoride group was significantly lower than low fluoride group (P ＜ 0.05).The levels of serum slL-6R in control group,low fluoride group,middle fluoride group and high fluoride group were (0.83 ± 0.20),(0.93 ± 0.23),(0.82 ±0.27) and (0.92 ± 0.28) μg/L,respectively.The differences between groups were not statistically significant (F =0.738,P ＞ 0.05).Immunohistochemical results showed that articular cartilage full-layer cells in each group expressed IL-6 protein especially in the middle layer of chondrocytes,while IL-6 protein only expressed in hypertrophic chondrocytes of epiphyseal plate cartilage.Comparing with other groups,IL-6 positive cells were the most and had the deepest staining in low fluoride group.Conclusions Different doses of NaF could not only cause cartilage lesion,but also change the expression of IL-6 in serum and cartilage tissue of Balb/c mice.The results indicate that IL-6 may be involved in the cartilage lesion caused by fluoride.