实验性氟中毒骨组织中Ⅱ型胶原基因表达及结构的改变  

Expression of type Ⅱ collagen gene and structural change in bone tissues of rats with experimental fluorosis

作　　者：刘秉慈[1] 许增禄[2] 缪庆[1] 徐园园[2] 徐茗[1] 钱晓菁[2] 尤宝荣[1] 袁伯华[1] 康宁[1] 

机构地区：[1]中国疾病预防控制中心职业卫生与中毒控制所,北京100050 [2]中国医学科学院基础医学研究所中国协和医科大学基础医学院

出　　处：《中华预防医学杂志》2003年第4期243-245,T002,共4页Chinese Journal of Preventive Medicine

基　　金：国家自然科学基金重点资助项目 (3 973 0 3 90 )

摘　　要：目的　观察过量氟对大鼠骨组织中Ⅱ型胶原基因表达的影响和对大鼠骨骼中胶原类型及胶原结构的作用。方法　制备Ⅱ型胶原cDNA探针 ,采用cDNA mRNA原位杂交技术 ,检测饮用过量氟水 (氟化钠 2 2 1mg/L)的大鼠骨组织中Ⅱ型胶原的改变。采用苦味酸天狼星红染色 偏振光法对骨组织中胶原类型及胶原纤维结构进行观察。结果　染氟后 ,股骨等骨组织出现成软骨细胞。cDNA mRNA原位杂交显示 ,软骨化的骨组织软骨陷窝的软骨胞质中可见Ⅱ型胶原基因表达。染氟0 5个月组肋软骨软骨细胞胶原mRNA水平最高 ,其灰度值和标准误分别为 0 0 86和 0 0 13,染氟 1个月及 2个月后逐渐减弱 ,其灰度值和标准误分别为 0 0 4 7、0 0 0 8和 0 0 39、0 0 0 7。苦味酸天狼星红染色 偏振光法观察骨组织中出现多色性Ⅱ型胶原纤维 ,胶原纤维断裂、排列紊乱、含量下降。结论　过量氟可致骨组织中胶原类型和胶原结构的改变 ,导致Ⅱ型胶原在软骨化骨组织中的表达及在肋软骨组织中的表达增强。Objective To investigate the effects of excessive intake of fluoride on the expression of type Ⅱ collagen gene and types and morphological change of collagen fiber in the bone tissues of rats. Methods A rat model with fluorosis was established by adding 221 mg/L of sodium fluoride (NaF) to drinking water for the rats for 15 days, 30 days and two months, respectively. Type Ⅱ collagen α1(Ⅱ) cDNA probe was prepared, and cDNA-mRNA in-situ hybridization was employed to detect change in expression of type Ⅱ collagen mRNA in the bone tissues of rats with excessive intake of fluoride (221 mg/L NaF). Picrosirus-polarization method was used to observe types of collagen and morphology of collagen fiber in the bone tissues. Results Chondroblasts were found in the femur and other bone tissues of the rats after exposure to fluoride. cDNA-mRNA in-situ hybridization showed that expression of type Ⅱ collagen gene could be observed in the cytoplasm of chondrocytic lacuna and chondrified bone tissues. mRNA in collagen of chondrocytes of the rib cartilage reached the peak level 15 days after exposure to fluoride, and decreased gradually one month and two months after exposure. Polychromatic type Ⅱ collagen, breakage of collagen fiber, disorder array and reduced content of type Ⅱ collagen could be found in the bone tissues with picrosirius-polarization method. Conclusions Excessive intake of fluoride could lead to changes in types and structure of collagen (cross-linkage) of bone tissues, which caused expression of type Ⅱ collagen gene in the chondrified bone tissues and enhanced its expression in the rib cartilage tissues.

关 键 词：实验性氟中毒 骨组织 Ⅱ型胶原 基因表达 胶原纤维结构 cDNA-mRNA 原位杂交技术 

分 类 号：R114[医药卫生—卫生毒理学]