补肾法对酒精干预人成骨样细胞OS-732蛋白质表达的影响  

Influence of replenishing-kidney therapy on protein expression of human osteoblast-like cell OS-732 intervened by alcohol

作　　者：刘武[1] 樊粤光[2] 夏雄智[3] 曾毅荣[2] 刘建仁[2] 王海彬[2] 

机构地区：[1]广西中医学院第一附属医院骨科,广西530023 [2]广州中医药大学第一附属医院骨科 [3]广东省第二中医院骨科

出　　处：《北京中医药大学学报》2009年第9期614-617,共4页Journal of Beijing University of Traditional Chinese Medicine

基　　金：高等学校博士学科点专项科研基金项目(No.20060572002)

摘　　要：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 intervened by alcohol, and therapeutic effects of Chinese herbal remedies with replenishing-kidney action on these changes Methods New Zealand rabbits （n = 14 ） were randomly divided into the treatment group given Liuweidihuang Pills intragastrically （5.4 g dissolving in 10 mL warm normal saline solution） according to the conversion rate between rabbit and human, and blank control group given normal saline solution in the same quantity, once a day for 3 days repectively. The blood samples were collected from rabbits 2 hours after the last time administration of remedies （double quantity） to prepare the medicated serum. The serum then was mixed with RPMI1640 cell culture medium and made up the culture solution with the concentrations of 5%, 10% and 20% respectively, in which osteoblast-like cells were cultured. The best concentration of medicated serum （containing Liuweidihuang Pills） was 10% through the observation on cell growth curve. The osteoblast-like cells OS-732 were divided into the alcohol intervened group, treatment group and blank control group. After cell culture and collection the total protein was extracted from RPMI 1640 cells in every group and separated by using dimensional gel electrophoresis technique, and the gel electrophoresis atlas of protein spots were obtained, which then were analyzed and the different protein spots were found out by computer image analysis software. Results After comparing among three groups there were 5 protein spots with significant difference （i. e. SSP1002, SSP1401, SSP1602, SSP8501 and SSP8504）. The expressions of protein spots in the alcohol intervened group were lower than those in the blank control group. The above 5 protein spots were reversed in different extent when the treatment group was compared with the alcohol intervened group. Conclusion The Chinese herbal remedies with replenishing-kidney action may enhance the protein expression of injured osteoblast-like cells.

关 键 词：酒精 成骨样细胞 蛋白质表达 补肾法 兔 

分 类 号：R285.5[医药卫生—中药学]