【摘要】 目的观察1,25二羟维生素D31,25(OH)2D3诱导成年大鼠骨髓来源的单核细胞形成多核破骨细胞的作用。方法实验分为单核细胞诱导组和全骨髓诱导组,每组各10只大鼠。密度梯度离心法分离大鼠骨髓中的单核细胞,α-MEM培养液中加入10-8mol/L的1,25(OH)2D3诱导单核细胞向破骨细胞分化。利用抗酒石酸酸性磷酸酶(TRAP)染色、倒置相差显微镜和扫描电镜,观察诱导不同时相TRAP阳性(TRAP+)多核细胞的形态学特征、特异性酶表达、以及骨吸收陷窝。进而计数诱导的破骨细胞数量,鉴定其功能活性,并与全骨髓诱导方法比较。结果骨髓单核细胞诱导组在培养第7d出现TRAP+多核细胞,第14dTRAP+多核细胞数量达峰值,第21d TRAP+多核细胞数量开始减少。从培养第14d到第23d,单核细胞诱导组TRAP+多核细胞数量多于全骨髓诱导组(P<0.05);培养第10d到第23d单核细胞诱导组骨吸收陷窝数量多于全骨髓诱导组(P<0.05)。单核细胞诱导组破骨细胞数量高峰期可持续7d,全骨髓诱导法仅持续3d。结论1,25(OH)2D3能够诱导成年大鼠骨髓来源的单核细胞形成破骨细胞;所诱导的破骨细胞的数量和峰值持续时间、以及骨吸收活性均高于全骨髓诱导方法。更多还原

【Abstract】 Objective To investigate the effect of 1,25(OH)2 dihydroxyvitamin D3 on the generation of osteoclasts from mononuclear cells of adult rats. Methods With density gradient centrifugation, the mononuclear cells were isolated from rat bone marrow and cultured in the α-MEM with 10-8 mol/L 1,25(OH)2 dihydroxyvitamin D3. The induced cells were fixed and stained with tartrate-resistant acid phosphatase (TRAP). The bone resorption pits were examined by scanning electron microscope, and the morphology of osteoclasts were examined with inverted phase contrast microscope. Results The TRAP+ multinucleated cells could be observed on the seventh culturing day of mononuclear cell group. The number of TRAP+ multinucleated cells went up to a peak on the fourteenth culturing day, and then began to decrease on the twenty-first culturing day. The number of osteoclasts induced from mononuclear cell group was more than that from bone marrow group (P<0.05) when it was during the fourteenth culturing day to the twenty-third culturing day. The number of bone resorption pits in the mononuclear cell group was much more than that in the bone marrow group (P<0.05) when it was in period from the tenth culturing day to the twenty-third culturing day. The number of osteoclasts induced from mononuclear cell group could keep the peak value of lasting 7 days, but that induced from bone marrow group could only last its peak value for 3 days. Conclusion The method with 1,25(OH)2 dihydroxyvitamin D3 inducing the formation of osteoclasts from the marrow mononuclear cells is better than that from the whole bone marrow.更多还原