慢性氟中毒大鼠骨组织形态计量学改变及血清骨代谢标志物表达

曾贝贝; 陈修文; 王楠兰; 吴彦秋; 吴延莉; 夏敏; 张业芳; 张华; 韦艳

doi:10.13421/j.cnki.hjwsxzz.2016.02.001

慢性氟中毒大鼠骨组织形态计量学改变及血清骨代谢标志物表达

Histomorphometric Change of Bone and Metabolic Marker Expression in Rats Induced by Chronic Fluorosis

摘要

摘要:
目的探讨慢性氟中毒大鼠骨组织病理形态学改变及与氟性骨损伤相关的骨代谢标志物的改变。

方法 48只健康SD大鼠随机分为4组,对照组饮用自来水,低氟组、中氟组、高氟组分别饮用含50 mg/L、150 mg/L和250 mg/L氟化钠的自来水。饲养6个月后,检测氟斑牙发生率,氟离子选择电极法检测各组大鼠尿氟、骨氟、血氟含量;取大鼠股骨干骺端进行苏木精-伊红染色,光镜观察骨组织形态学改变;酶联免疫吸附法(ELISA)检测大鼠血清骨代谢标志物含量。

结果大鼠体重、氟斑牙发生情况与氟负荷量的变化:低中氟组雌鼠体重增幅较对照组小,高氟组雌鼠体重增幅较对照组大;高氟组雄鼠体重增幅小于对照组、低、中氟组雄鼠。各染氟组大鼠氟斑牙检出率、尿氟、骨氟和血清氟含量均高于对照组,差异有统计学意义(P<0.05),而且随染氟剂量增高,氟斑牙严重程度、尿氟、骨氟和血清氟呈上升趋势。大鼠股骨组织形态计量学结果:中、高氟组雌雄大鼠的骨皮质厚度均高于对照组,差异有统计学意义(P<0.05),中、高氟组雄鼠骨小梁面积大于对照组,差异有统计学意义(P<0.05);低氟组雌雄鼠骨小梁平均面积均小于对照组,差异有统计学意义(P<0.05);低、中、高氟组雌雄成骨细胞数均低于对照组,差异有统计学意义(P<0.05)。BGP、BALP、PⅠNP检测结果:低、中、高氟组雌雄鼠BGP、BALP、PⅠNP均小于对照组,差异多有统计学意义(P<0.05)。CTX-Ⅰ、TRACP-5b检测结果:低、中、高氟组雌雄鼠CTX-Ⅰ、TRACP-5b均小于对照组,差异多有统计学意义(P<0.05)。

结论经饮水染氟6个月后,反映成骨活性和破骨活性的骨代谢指标在不同剂量组中表现为不同程度下调,骨组织形态计量显示中高氟组大鼠骨量增加;低氟组大鼠则出现骨量减少的指征。提示氟对成骨活性与破骨活性均有影响,且与染氟剂量有关。

Abstract:
Objectives In order to investigate the patho-morphological change in the bone tissue of rats caused by chronic fluorosis and the change of metabolic marker of bone injury related to fluorosis.

Methods Forty eight healthy SD rats were randomly divided into 4 groups, including a control group and 3 fluoride treated groups, the concentration of sodium fluoride in their drinking water was 0 mg/L, 50 mg/L, 150 mg/L and 250 mg/L, respectively. After six months of exposure to fluoride, the incidence of dental fluorosis and the content of fluoride in urine, bone and blood were detected by fluoride ion selective electrode method. The femoral metaphysis of rat was stained with hematoxylin eosin (HE) and observed under optical microscope. The levels of bone metabolic marker in serum were determined by enzyme-linked immunosorbent assay(ELISA).

Results The gain of body weight was less in fluoride treated groups than that in the control group except in the high fluoride female group. The incidence of dental fluorosis and the level of fluoride in urine, bone and blood in treated groups were higher than those in the control group (P<0.05), the degree of dental fluorosis was correlated with the increase of fluoride in drinking water and the extension of experimental time. The cortical thickness of femur bone in medium and high fluoride group was higher than that of control group (P<0.05). The trabecular area in medium and high fluoride group was higher than that of the control group(P<0.05), but the trabecular area in low fluoride group was less than the control group (P<0.05); The number of osteoblasts in cortical bone of 3 treated groups were lower than that of the control group(P<0.05). Compared with the control group, the level of BALP, BGP and PⅠNP in 3 treated groups were less than that of the control group (P<0.05); and the level of CTX-Ⅰand TRACP-5b in most of treated groups were less than the control group (P<0.05).

Conclusions After fluoride being added into drinking water for 6 months, the metabolic indexes reflecting the activity of osteoblast and osteoclast were down-regulated. The bone mass of rats exposure to 150 mg/L and 250 mg/L NaF in drinking water was increased, while those exposure to 50 mg/L NaF was decreased under histomorphometric examination. It suggested that fluoride exert an impact on the activity of osteoblast and osteoclast, and the effect was related to the dosage of. fluoride.

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