摘要
为研究补骨脂对体外成骨细胞增殖的影响。以出生 2 4h以内的wistar大鼠为来源 ,酶消化法分离培养新生大鼠颅盖骨成骨细胞 ,胰蛋白酶消化后 ,传代培养大鼠成骨细胞 ;取一定量接种于 2 4孔培养板细胞贴壁后 ,分成实验组 1 5与空白对照组 ,分别用含补骨脂注射液 5 %、12 .5 %、2 5 %、5 0 %、10 0 %、0 %的DMEM培养基进行培养 ,连续观察 7天 ,绘制细胞生长曲线 ;另取一定量接种于 96孔培养板 ,分组及加入培养基同上 ,MTT法观察细胞存活和增殖能力。结果显示原代成骨细胞逐渐形成成纤维细胞集落 ,有成纤维细胞型表现 ;传代成骨细胞逐渐被细胞分泌的基质所包埋 ,形成钙化。补骨脂对新生大鼠成骨细胞的增殖有显著促进作用。表明利用胰酶消化法可以分离培养出纯度高 ,数量较多且培养时间明显缩短的成骨细胞。补骨脂抗骨质疏松的作用与其增加成骨细胞的数量和促进成骨细胞的增殖能力有关。
The objective of the paper is to study the effect of psoralea injection on invitro-cultured rat osteoblasts proliferation. Taking neonatal Wister rats aged within 24 h as the source, cranial osteoblasts were isolated and cultured with enzyme digestion. After trypsin digestion, the rat osteoblasts were cultured generatively; a proper amount of third-generation osteoblasts were inoculated into 24-hole culture plates and divided into the experimental groups I-V and the blank group, which were cultured in the DMEM media containing 5 %, 12.5 %, 25 %, 50 %, 100 % and 0 % of psoralea injection, respectively, observed successively for 7 days, and the cell growth curves were drawn; another proper amount of third-generation osteoblasts were inoculated into 19-hole culture plates, divided into the same groups, and cultured in the same media, as the above, the cell survival and the reproduction ability were observed with MTT method. The results showed that the primary osteoblasts gradually became fibroblastic colonies having the fibroblastic type expression; the generated osteoblasts were gradually embedded by the cell-secreted medium, forming calcification. Psoralea has a significantly stimulating effect on newborn rat osteoblasts proliferation. Tryptic digestion could isolate and culture numbers of osteoblasts with a high purity and obviously shorten the culture time. The anti-osteoporosis effect of psoralea may be related to its function to increase the number of osteoblasts and accelerate osteoblasts proliferation.
出处
《中医正骨》
2004年第6期6-7,共2页
The Journal of Traditional Chinese Orthopedics and Traumatology
