摘要
目的观察不同剂量氟对BALB／c小鼠胫骨和腰椎骨量的影响。方法采用随机数字表法按体质量将64只4周龄雄性BALB／c小鼠分为对照组及低、中、高氟组，每组16只，分别饮用含氟0（对照）、25、50、100mg／L的氟化钠（NaF）水溶液，饲喂12周后建立氟中毒小鼠模型。染氟结束后，观察各组小鼠氟斑牙发病情况，并取小鼠血液、两后肢、脊柱。采用氟离子选择电极法检测小鼠骨氟；微量酶标法检测血清碱性磷酸酶（AKP）及酸性磷酸酶（ACP）活性；透射电镜观察腰椎骨组织中成骨细胞、破骨细胞的超微结构；光镜下观察各染氟组小鼠胫骨、腰椎骨小梁的病理形态学改变，并用lmage-ProPlus（IPP）软件计算骨小梁面积百分数（％，Tb．Ar）来反应骨量。结果低、中、高氟组小鼠骨氟水平[（1828．62±102．93）、（3308．27±185．63）、（4933．36±301．16）mg／kg]均高于对照组[（775．23±92．56）mg／kg，P均〈0．05]。对照组，低、中、高氟组小鼠氟斑牙检出率分别为0（0／16）、47％（7／15）、93％（14／15）、100％（16／16），组间比较差异有统计学意义（x^2=27．23，P〈0．05）。中、高氟组小鼠血清AKP[（18．30±1．99）、（24．50±3．14）金氏单位／100m1]、ACP[（11．97±1．73）、（11．31±1．46）金氏单位／100m1]的活性均高于对照组[（14．63±1．21）、（9．07±1．47）金氏单位／100ml，P均〈0．05]。电镜下各染氟组成骨细胞细胞器发达，含有丰富的粗面内质网和高尔基体、线粒体，核仁明显，有些成骨细胞侧面有突起与邻近成骨细胞相连，处于功能活跃期；低、中氟组破骨细胞线粒体丰富，皱褶缘清晰，并且分布有吞噬泡，功能也较活跃。光镜下小鼠胫骨石蜡切片苏木素．伊红（HE）染色显示，高氟组骨小梁％Tb.Ar（28．79±8．26）与对照组（17．03±3．73）相比明显增加，差异有统计学意义（P〈0．05）；小鼠腰椎石蜡切片HE染色显示，低、中、高氟组骨小梁％Tb-Ar（15．87±2．59、18．28±0．89、21．99±1．81）均高于对照组（12．06±1．76，P均〈0．05）。结论应用BALB／c小鼠可以成功建立氟中毒的动物模型，氟所致BALB／c小鼠氟性骨损伤表现为成骨、破骨均活跃，但以成骨活性增强为主．骨形成大于骨吸收，骨量增加，并且在小鼠腰椎骨组织中表现更加明显。
Objective To investigate the effects of fluoride on trabecular bone of the tibia and lumbar in BALB/c mice. Methods Totally 64 four-week-old BALB/c mice were randomly divided into 4 groups by weight, 16 per group： control group （treated with distilled water） and 3 sodimn fluoride （NaF） exposure groups （treated with NaF at 25, 50 and 100 mg/L F-）, respectively. At 12 weeks, mice were killed and blood, two hind limbs and lumbar were collected. Bone fluoride content and incidence rates of dental fluorosis were determined. Serum content of alkaline phosphatase （AKP） and acid phosphatase （ACP） were detected by micro enzyme labeled method. The uhrastructure of osteoblasts and osteoclasts in lumbar were observed via transmission electron microscope. The pathological changes of the trabecular bone of the tibia and the lumbar were observed under optical microscope, the percentage of trabecular area （%Tb.Ar） was measured with Image-Pro Plus （IPP） software. Results Bone fluoride contents of low, middle and high fluoride groups [（1 828.62 ± 102.93）, （3 308.27 ± 185.63）, （4 933.36 ± 301.16） mg/kg] were higher than that of the control group [（775.23 ± 92.56） mg/kg, all P 〈 0.05]. The incidences of dental fluorosis in the 4 groups were 0（0/16）, 47% （7/15）, 93% （ 14/15 ） and 100% （16/16）, respectively; the difference was statistically significant （x^2 = 27.23, P 〈 0.05）. In middle and high fluoride groups, serum AKP [（18.30 ± 1,99）, （24.50 ± 3.14） king unit/100 ml] and ACP [（11.97 ± 1.73）, （11.31 ± 1.46） king unit/100 ml] were significantly higher than those of control [（14.63 ± 1.21）, （9,07 ± 1.47） king unit/100 ml, respectively, all P 〈 0.05]. Under the electron microscope, osteoblast had developed organelles in each fluoride group, rough endoplasmic retieulum, Golgi body, and mitochondria were abundant, and nucleolus was obvious in the osteoblast. Osteoclast was rich in mitoehondria, ruffled border clear and distributed phagocytic vacuoles in low fluoride group and middle fluoride group. Compared with the control group （17.03 ± 3.73）, HE staining of tibia %Tb.Ar in high fluoride group （28.79 ± 8.26） was significantly increased （P 〈 0.05）. The lumbar spine %Tb.Ar in low, middle and high fluoride groups （15.87 ± 2.59, 18.28 ± 0.89, 21.99 ± 1.81） were higher than that of the control group （12.06 ± 1.76, all P 〈 0.05]. Conclusions BALB/e mice could be used as a model of skeletal fluorosis. Osteoblast and osteoelast are activated in BALB/c mice with skeletal fluorosis. Bone formation is more obvious than bone resorption and bone mass is increased. What is more, bone mass has increased more significantly in the lumbar spine of mice.
出处
《中华地方病学杂志》
CAS
CSCD
北大核心
2017年第4期235-240,共6页
Chinese Journal of Endemiology
基金
黑龙江省科学基金项目（JC2015018）
关键词
氟
骨和骨组织
成骨细胞
破骨细胞
Fluorides
Bone and bone tissue
Osteoblast
Osteoclast
