冰蜜生肌膏对兔成纤维细胞增殖影响的实验研究

Research of immunohistochemistry on the influence of borneol and honey ointment(BHO)promote proliferation of the fibroblasts of rabbits

目的：观察冰蜜生肌膏促进体外培养兔成纤维细胞增殖的影响.方法：采用消化分离法进行兔成纤维细胞的原代培养,通过形态学和免疫组织化学的方法鉴定成纤维细胞.第3代纯化的成纤维细胞分为5组,空白对照组每孔加入等量的PBS,实验组每孔分别加入125、62.5、31.25、15.625 mg/ml的冰蜜生肌膏提取液进行培养,培养24、48、72 h后分别行MTT法检测每孔的光吸收值（OD值）.结果：冰蜜生肌膏提取液加入到成纤维细胞中培养24、48、72 h后,经MTT法检测,不同浓度（浓度分别为125、62.5、31.25、15.625 mg/ml）冰蜜生肌膏与对照组OD值比较差异有显著性统计学意义（P＜0.05）.结论：15.625 mg/ml浓度冰蜜生肌膏对成纤维细胞的生长有明显促进作用（P＜0.05）,药物浓度在15.625～62.5 mg/ml时其作用随药物浓度升高而增加,并在浓度为62.5 mg/ml时达到最大效应值（P＜0.01）,当药物浓度大于62.5 mg/ml时则作用趋向于饱和,提示一定浓度范围内的冰蜜生肌膏药液能明显增加成纤维细胞数量,且促进作用和浓度呈显著依赖性.

Objective ： To observe the influence of BHO on the promotion of rabbit fibroblasts proliferation. Methods：Fibroblasts of rabbit were obtained from the digested and aparted method of primary culture and identified by morphology and immunohistochemistry. The third purified fibroblasts were divided into 5 groups. Each hole of the blank compared group was added into same quantity PBS to culture. Each hole of the experimental groups was cultured respectively with the concentration of BHO of different quantity（ 125,62. 5,31.25, 15. 625 mg/ml）. MTT method was used to measure OD number of each hold after being cultured for 24,48, 72 h respectively. Results：Fibroblasts of rabbit were obtained from technology of cell culture and identified by morphology and immunohistochemistry. Compared with blank groups, OD numbers with different concentration BHO were obviously different （P 〈 0. 05 ）. Conclusion： ①Fibroblasts of rabbit could be obtained from the digested and aparted method of primary cell cluture;②BHO of regular concentration could increase the numbers of fibroblast obviously.