N-脱硫酸肝素对ICR雌性小鼠骨形态计量学的影响

Effect of N-desulfated heparin on bone morphometry in ICR female mice

目的探讨肝素诱发骨质疏松症的发病机理。方法ICR雌性小鼠共30只分成3个实验组,普通肝素组和N-脱硫酸肝素组各10只,分别按1IU/g.day给予药物;生理盐水组10只,给予同体积的生理盐水。自12w月龄开始每日皮下注射,28d后心脏采血,检测PT及APTT;取双侧胫骨,制成组织切片,ALP和TRAP染色计数成骨细胞和破骨细胞;使用图形处理软件测量兴趣区域(AOI)的骨小梁长度和面积等,结果进行t检验。结果普通肝素组与N-脱硫酸肝素组在骨小梁面积、成骨细胞计数、破骨细胞计数方面两组之间比较无显著性差异,而与生理盐水组之间存在显著性差异(P<0.05);N-脱硫酸肝素组与普通肝素组在PT、APTT方面均存在显著性差异(P<0.05)。结论N-硫酸基团在普通肝素的体内抗凝血作用中起重要作用,但是该基团与诱导骨质疏松的形成之间缺乏明显的相关性。

Objective To investigate the pathogenesis of osteoporosis induced by heparin. Methods Thirty ICR female mice were divided into three groups： the regular heparin, the N-desulfated heparin and the control group （10 mice of each）. The first two groups were given subcutaneous regular or N-desnlfated heparin at 1 U/g/d, respectively, while the control was given normal saline at the same dose only. The injection was started since 12 weeks of age. Four weeks later, the blood in the heart was obtained and the levels of PT and APTT were examined. The samples of tibia of both sides were taken and the amount of osteoblasts and osteoclasts were calculated after ALP and TRAP staining. The length and area of bone trabecular in AOI were measured with the graph-disposal software.Results Significant difference was shown in the area of bone trabecular, the amount of osteoblasts and osteoclasts between the two heparin groups and the control（ P 〈 0.05）, but not within the two heparin groups（ P 〉 0.05）. The regular heparin group showed significant difference in the level of PT and APTT compared with the N-desulfated heparin group （ P 〈 0.05 ）. Conclusions N-sulfuric acid radical plays an important role in the anticoagulant function of regular heparin in vivo and is not associated with inducing osteoperosis.