乙二胺四亚甲基膦酸钙预防性给药对卵巢切除大鼠骨质疏松的作用

Effects of preventive administration of calcium ethylenediamine tetramethylene phosphonate on osteoporosis rats induced by ovariectomy

目的探讨预防给乙二胺四亚甲基膦酸钙(Ca-EDTMP)对卵巢切除大鼠骨质疏松的作用。方法 SD雌性大鼠80只,随机分成8组:假手术组、模型组、Ca-EDTMP2,4,8,16,32mg·kg-1组和氯膦酸二钠80mg·kg-1阳性药对照组。通过切除卵巢的方法制备大鼠骨质疏松症模型。手术后第4天开始给药,Ca-EDT-MP组大鼠每4周iv给予1次,共4次;阳性对照组po给予氯膦酸二钠50mg·kg-1,每天1次,连续120d。末次给药后1周处死大鼠,分别采用甲基麝香草酚蓝法和紫外法检测血清Ca和P水平,AMP法碱性磷酸酶(ALP)活性;Holigic骨密度测定仪测定大鼠右股骨骨密度;采用甲基麝香草酚兰法和紫外法检测骨组织中Ca和P含量;采用生物力学测定仪检测弹性模量、最大载荷、最大挠度、弹性压力和弹性应变;HE染色检查股骨骨小梁的变化。结果与模型组比较,Ca-EDTMP2,4,8,16和32mg·kg-1组血清Ca在正常范围内波动,而血清P水平有明显下降,分别降低了8.6%,11.5%,11.1%,13.8%和10.0%(P<0.05,P<0.01);ALP活性分别下降了5.9%,6.9%,9.3%,12.0%和14.3%(P<0.05,P<0.01);Ca-EDTMP2和4mg·kg-1组能增加卵巢切除大鼠股骨骨密度,但无统计学差异;Ca-EDTMP8,16和32mg·kg-1组股骨骨密度提高了7.3%,8.2%和12.5%(P<0.05,P<0.01)。Ca-EDTMP4,8,16和32mg·kg-1组骨组织中Ca和P均明显增加,Ca分别增加了1.9%,2.2%,2.7%和3.3%(P<0.05,P<0.01),P含量分别增加了1.1%,2.3%,3.4%和4.6%(P<0.05,P<0.01)。Ca-EDTMP2,4,8,16和32mg·kg-1组最大载荷显著增加,分别增加了23.9%,27.2%,28.7%,45.7%和37.2%(P<0.01);Ca-EDTMP8,16和32mg·kg-1组的弹性变化显著增加,分别增加了44.8%,39.2%和45.2%(P<0.05)。Ca-EDTMP32mg·kg-1组骨密质厚度正常,细胞层次较清晰,骨小梁增宽,相互联结,骨髓较丰富。结论预防性给予Ca-EDTMP可防止骨量丢失,抑制骨吸收,对防治骨质疏松有一定作用。

OBJECTIVE To investigate the effect of calcium ethylenediamine tetramethylene phosphonate(Ca-EDTMP) on osteoporosis rats induced by ovariectomy.METHODS Eighty Sprague-Dawley(SD) female rats were randomly divided into 8 groups:sham,model,Ca-EDTMP 2,4,8,16 and 32 mg·kg-1 and positive drug clodronate disodium groups.The rats were given drugs on the 4 th day after operation.Ca-EDTMP was iv given once every four weeks for totally four times.Clodronate disodium was given po 50 mg·kg-1 daily for 120 d.The rats were executed 1 week after the last administration.Serum Ca,P level and alkaline phosphatase(ALP) activity,bone mineral density(BMD) of the right femur,Ca and P content in bone tissue,bone biomechanics indicators including elastic modulus,maximal loading,maximal bending,elastic stress and elastic strain,and femoral trabecular bone changes were detected.RESULTS Compared to model group,the serum Ca level of Ca-EDTMP groups fluctuated in the normal range and P level in serum decreased by 8.6%,11.5%,11.1%,13.8% and 10.0%(P < 0.05,P <0.01) ;ALP activity in serum decreased by 5.9%,6.9%,9.3%,12.0% and 14.3%(P <0.05,P <0.01).BMD in Ca-EDTMP 8 and 16 and 32 mg·kg-1 groups significantly increased by 7.3%,8.2% and 12.5%(P < 0.05,P <0.01).Bone mineral content of Ca in Ca-EDTMP 4 and 8 and 16 and 32 mg·kg-1 groups increased by 1.9%,2.2%,2.7% and 3.3%(P < 0.05,P <0.01) ;content of P increased by 1.1%,2.3%,3.4% and 4.6%(P < 0.05,P < 0.01).Maximal loading in Ca-EDTMP groups increased by 23.9%,27.2%,28.7%,45.7% and 37.2%(P < 0.01).Elastic change of Ca-EDTMP 8 and 16 and 32 mg·kg-1 groups increased by 44.8%,39.2% and 45.2%(P <0.05).The thickness of compact bone substance of the rats in Ca-EDTMP 32 mg·kg-1 group was normal and delamination of cells was clear and bone trabecula was broadened and marrow was plentiful.CONCLUSION Ca-EDTMP could prevent bone loss and inhibit bone resorption,potentially effective for osteoporosis prevention and treatment.