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目的观察成骨样细胞在酒精干预后的变化和补肾中药对其的治疗作用。方法首先用新西兰兔14只,随机分配成中药六味地黄丸治疗组和生理盐水空白对照组。依据人与兔关系换算出治疗组每日用药5.4 g,用温生理盐水10 mL融解灌胃;空白对照组灌胃等量生理盐水,1次/d,连续3 d,末次双倍量灌胃2 h后采血制备含药血清。然后将含药血清与RPMI 1640细胞培养基配成浓度为5%、10%、20%培养液,对细胞进行培养,通过观察细胞生长曲线,得出最佳含药血清浓度为10%。最后将人成骨样细胞OS-732分为酒精干预组、药物治疗组、空白对照组进行培养并收集细胞。每组细胞提取总蛋白后进行双向凝胶电泳技术分离,得出蛋白质点电泳凝胶图谱,利用计算机图像分析软件分析找出差异蛋白质点。结果酒精干预组、药物治疗组、空白对照组比较,表现明显差异的蛋白点有5个(即SSP1002、SSP1401、SSP1602、SSP8501、SSP8504),酒精干预组与空白组比较蛋白点表达均降低;药物治疗组与酒精干预组比较,这5个蛋白点表达均得到不同程度的逆转。结论补肾中药可能能够提高受损成骨细胞的蛋白表达。
Objective To observe the changes of osteoblast-like cells after alcohol intervention and the therapeutic effects of tonifying kidney on them. Methods First of all, 14 New Zealand rabbits were randomly assigned to a Chinese herbal medicine Liuweidihuangwan treatment group and a normal saline control group. According to the relationship between human and rabbit, the treatment group was treated with 5.4 g daily, dissolved in warm saline 10 mL, and the blank control group was given normal saline once a day for 3 days. The last double-dose intragastric administration Blood was collected after 2 h to prepare medicated serum. The drug-containing serum was then mixed with RPMI 1640 cell culture medium to a concentration of 5%, 10% and 20%, and the cells were cultured. The optimal serum concentration was found to be 10% by observing the cell growth curve. Finally, human osteoblast-like cells OS-732 were divided into alcohol intervention group, drug treatment group, blank control group for culture and collection of cells. Total proteins were extracted from each group and then separated by two-dimensional gel electrophoresis. The gel electrophoresis of protein spots was obtained and analyzed by computer image analysis software to find out the differences of protein spots. Results There were 5 protein spots (SSP1002, SSP1401, SSP1602, SSP8501, SSP8504) in alcohol-treated group, drug-treated group and blank control group. The protein spots in alcohol-treated group and blank group were lower than those in control group Treatment group and alcohol intervention group, the expression of these five protein spots were different degrees of reversal. Conclusion Bushen Chinese medicine may be able to improve the protein expression of impaired osteoblasts.