龟鹿二仙胶汤含药血清对兔骨髓基质干细胞增殖的影响  

Effects of"Guilu Erxian Jiao Decoction"Serum on Proliferation of Bone Marrow Stromal Cells in Rabbits in Vitro

作　　者：李文顺[1] 李楠[2] 王和鸣[2] 李西海[2] 詹红生[1] 石印玉[1] 王拥军[3] 周重建[3] 施杞[3] 李月平 王永强[5] 张林[6] 

机构地区：[1]上海中医药大学附属曙光医院骨伤科,上海200021 [2]福建中医学院骨伤系,福建福州350003 [3]上海中医药大学附属龙华医院骨伤科,上海200032 [4]山东省东营市胜采医院骨科,山东东营257230 [5]山东中医药高等专科学校骨伤系,山东烟台264100 [6]复旦大学药学院,上海201203

出　　处：《上海中医药大学学报》2009年第1期56-60,共5页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：教育部高等学校博士学科点专项基金资助项目(20050393002);国家自然科学基金资助项目(30772814);福建省教育厅科研基金资助项目(JA04212)

摘　　要：目的:观察龟鹿二仙胶汤对骨髓基质干细胞(BMSCs)增殖的影响,确定影响BMSCs增殖的最佳中药血清浓度。方法:经前期体外分离、纯化、培养获得兔BMSCs,运用不同浓度的龟鹿二仙胶汤中药含药血清对获得的兔BMSCs进行药物干预24、48、72 h后,采用MTT、免疫组化、流式细胞仪等方法,研究龟鹿二仙胶汤对BMSCs增殖的影响。结果:MTT比色实验显示,龟鹿二仙胶各浓度组均能不同程度地促进BMSCs的能量代谢,促进BMSCs增殖,与空白对照组相比有显著差异(P<0.01),而且以15倍药物浓度组为最佳。免疫组化结果证实,龟鹿二仙胶汤含药血清可通过促进PCNA表达来控制DNA复制,以15倍龟鹿二仙胶汤药物浓度血清培养时BMSCs表达致密的棕褐色颗粒最多,其他浓度组及TGF-β_3组较多,空白对照组则最少。FCM结果显示,二仙胶汤药物血清能显著提高BMSCs增殖指数,且以15倍药物浓度龟鹿血清培养时最明显。结论:龟鹿二仙胶汤刺激BMSCs增值与药物浓度有关,低浓度龟鹿二仙胶汤就能显著刺激BMSCs增殖,无论是48 h、72 h测验均为15倍药物浓度时具有显著的增值作用,而更高浓度龟鹿二仙胶汤则表现为促进BMSCs增殖作用减弱。Objective ： To investigate the effects of ＂Guilu Erxian Jiao Decoction＂ （GLEXJD） on proliferation of bone marrow stromal cells （BMSCs） and the optimum serum concentration, netheds：The BMSCs were isolated from the nucleated cells fraction of rabbit bone marrow with density gradient centrifuge and were extracted, purified, cultured in vitro. The BMSCs were interfered with（GLEX.ID） of different concentrations for 24, 48, 72 hours, and the proliferation of BMSCs were deceted with MTT assay, immunohistoehemistly and flow cytometry. Results：The MTT colorimetric assay showed that compared with the control group, the GLEXJD of different concentrations could promote the energy metabolism and the proliferation of BMSCs, with the optimum effect appeared at the 15 times of concentrations. Immunohistochemistry confirmed.that the GLEXJD serum could control the DNA duplication through promting the expression of PCNA, with the highest density of brown panicles occurred at the 15 times of concentrations. Among all the other groups, the density of brown particles in other administration groups and TGF-3 group were relatively higher than that of the control group. FCM showed that the GLEXJD drug serum could significantly increase the proliferation index of BMSCs, with the optimum effect appeared at the 15 times of concentrations. Conclution：The effect of GLEXJD on stimulating the proliferation of BMSCs is related to the drug serum concentration. GLEXJD can stimulate the proliferation of BMSCs even at the low concentration and the optimum effect appeared with 15 times of concentrations at the 48th, 72th hours. But the effect will be reduced with the drug serum concentration increasing even higher.

关 键 词：龟鹿二仙胶汤 骨髓基质干细胞 转化生长因子-Β3 含药血清 细胞增殖 

分 类 号：R285.5[医药卫生—中药学]