中药Liu-OGN对大鼠破骨细胞体外培养的影响

Effects of Liu-OGN on Rat Osteoclast Culture in Vitro

目的 :研究中药 Liu-OGN对体外培养的大鼠破骨细胞的作用。方法 :采用 Fenton[1] 等人的方法加以改良 ,从新生大鼠四肢长骨中分离培养破骨细胞 ,将破骨细胞均匀接种于含预置新鲜成年牛股骨骨磨片的 2 4孔培养板中 ;中药 Liu-OGN溶于 5 %阿拉伯胶中喂大鼠后 ,提取含中药 Liu-OGN药物血清加到培养液中 ,2 4孔培养板中的培养液每 2天更换一次 ,用不同药物干预 ,分别于培养期 2 4、4 8、72、96小时取出骨片TRAP染色 ,计数 TRAP( + )多核细胞数量、结果用 SPSS软件包分析。结果 :在培养期的前 4 8小时 ,每张骨片上 TRAP( + )的破骨细胞样细胞的数量有不同程度的增加 ,培养期 72小时后不同浓度中药 Liu-OGN对大鼠破骨细胞样细胞抑制作用明显 (与对照组相比 P<0 .0 1 )。结论 :中药 Liu-OGN对大鼠破骨细胞样细胞的体外生长有抑制作用 。

Objective:To study effect of chinese traditional medicine-Liu-OGN on rat osteoclasts.Methods:Using Fenton's method with some modification, we isolated and cultured osteoclasts from long bones of 1 day old wistar rats, and evenly inoculated on 24 well culture plates which put dentine slices in advance.Liu-OGN was dissolved in 5 percent Alrab gal and fed rats,then collected serum, put the serum into medium.Osteoclasts were treated by different drugs.The medium was repalced every 2 days.The culture was ended seperatively at 24?48?72?96hours and counted the number of TRAP positive multinucleated cells. The results were analysed by statistical package for social science(SPSS.)Results:The number of multinucleated TRAP positive osteoclast like cells present on the surface of each slice increated for first 48 hours. After 72 hours of culture, the number of osteoclast like cells was obviously inhibited by different concentration of Liu-OGN(compared to control P<0.01).Conclusion :Liu-OGN could inhibit rat osteoclast like cells growth in vitro, but this effect varied by concentraiton.