摘要
背景:中医"肾主骨"理论治疗激素性股骨头坏死是否对成骨细胞的蛋白表达变化起调节作用尚不清楚。目的:实验拟观察地塞米松对成骨样细胞的损伤及六味地黄丸的治疗作用机制,从蛋白水平观察激素引起成骨样细胞凋亡和中药对凋亡细胞保护作用的蛋白表达变化。设计、时间及地点:对比观察,于2005-06/2007-12在广州中医药大学分子生物实验室完成。材料:新西兰兔20只,雌雄各半,体质量(2.0±0.5)kg。方法:制备六味地黄丸含药血清,通过OS-732细胞增殖实验及碱性磷酸酶活性测定及引起OS-732细胞凋亡的实验分别筛选出中药及地塞米松的最佳剂量用于实验。实验分组:正常组用含体积分数为0.1小牛血清的RPMI1640培养液培养3d;模型组用高浓度(10-7mmol/L)地塞米松培养3d;治疗组在高浓度(10-7mmol/L)地塞米松培养2d后加入100g/L中剂量六味地黄丸含药血清3d。主要观察指标:提取正常组、模型组和治疗组OS-732细胞总蛋白的双向电泳图谱,对差异蛋白点进行分析。结果:模型组有7个蛋白表达下降,其中SSP1005、SSP2301、SSP3202和SSP65014个点最为明显,SSP5604点蛋白不发生表达;治疗组对上述7个点的表达均有不同程度的增强,SSP5604点明显增强。结论:糖皮质激素作用可以使成骨样细胞某些蛋白表达减弱,从而造成成骨样细胞凋亡;补肾中药可改善激素作用下OS-732细胞的蛋白表达变化,其作用是多靶点的。
BACKGROUND： According to the theory of ＂the kidney being in charge of bone＂, it is not clear whether the treatment of steroid-induced avascular necrosis of the femoral head cause the change of the protein expression on the osteoblasts. OBJECTIVE： To observe the action mechanism of Liuwei Dihuang Wan on treating the injury of osteoblast-like cells induced by dexamethasone （DX）. and to analyze the protein expression change of osteoblast-like cells apoptosis caused by hormone and the effect by using Chinese herbs. DESIGN, TIME AND SETTING： A controlled observation was performed in the laboratory of Molecular Biology of Guangzhou University of Chinese Medicine from June 2005 to December 2007. MATERIALS： Twenty New Zealand rabbits of either gender, weighing （2.0＋-0.5） kg, were adopted in this study. METHODS： The experimental serum containing Liuwei Dihuang Wan was prepared. The optimal doses of Chinese herbs and DX were defined by the trials for the OS-732 cells proliferation and apoptosis, as well as the detection of alkaline phosphatase activities. Normal control group was cultured for 3 days in RPMI 1640 culture solution containing 0.1 volume fraction of calf serum; Model group was cultured for 3 days with high dose of DX （10 7 mmol/L）; Treated group was cultured for 2 days with high dose of DX （10^- 7 mmol/L） and then continued culturing with middle dose （100 g/L） of Liuwei Dihuang Wan another for 3 days. MAIN OUTCOME MEASURES： The total protein of OS-732 cells cultivated in the normal control group, model group and treated group to analyze the differential protein spots by two-dimensional electrophoresis pattern. RESULTS： There were 7 protein expressions decreased in the model group, especially the spots of SSP1005, SSP2301, SSP3202 and SSP6501 decreased mostly. No protein expressed in the spot of SSP564; Protein expressing in the above-mentioned 7 spots were reinforced at different levels in the treated group, and that on the spot SSP5604 was reinforced mostly. CONCLUSION： Effect of glucocorticoid can attenuate some protein expressions of the osteoblast-like cells and cause apoptosis; Kidney-tonifying herbs exert a multitarget effect on ameliorating the protein expression of OS-732 cells affected by hormone.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第33期6411-6415,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
