杜仲叶对SD大鼠成骨细胞增殖及骨钙素表达水平的影响  

Effect of Eucommia Ulmoids Leaf on SD rat osteoblast proliferation and expression levels of osteocalcin

作　　者：方宁[1] 陈林攀[1] 邓鸣涛[1] 张立超[1] 杜川[1] 戴鹏[1] 刘荣华[2] 罗军[1] 

机构地区：[1]南昌大学第二附属医院,江西南昌330006 [2]江西中医药大学,江西南昌330006

出　　处：《时珍国医国药》2014年第11期2574-2576,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81160508)

摘　　要：目的研究不同浓度杜仲叶提取物对成骨细胞增殖及骨钙素表达的影响。方法取新生SD大鼠（〈24-48 h,6只）通过酶消化法分离出乳鼠原代成骨细胞;用碱性磷酸酶染色进行成骨细胞鉴定;先用无血清无酚红培养基饥饿SD大鼠成骨细胞2h后,再给予五组浓度梯度杜仲提取物（0,1,10,50,100 mg/ml）对SD大鼠成骨细胞进行干预、其中以浓度为0 mg/ml设为对照组,继续培养48 h后用MTT方法检测细胞增殖情况;同上步骤,用Western bolt分别检测五组浓度杜仲叶影响骨钙素（OC）的表达水平。结果碱性磷酸酶染色后的SD大鼠成骨细胞呈紫红色;MTT法结果显示：与对照组相比,杜仲叶提取物对成骨细胞的增殖具有促进作用（P〈0.05）,且具有浓度依赖性,其中浓度以50 mg/ml最为显著（P〈0.01）。Western bolt结果示各加药组与对照组比较,骨钙素蛋白表达水平均有显著的升高（P〈0.05）,同时也具有一致的浓度依赖性。结论杜仲叶提取物通过影响骨钙素表达对成骨细胞增殖具有促进作用,并具有浓度依赖性。Objective To study different concentrations of Eucommia extract effect on osteoblast proliferation and osteocalcin expression. Methods Newborn SD rats（ 24 - 48 h,6） are isolated by enzymatic digestion to get primary neonatal rat osteoblasts,osteoblasts were identified by alkaline phosphatase staining; First each groups were added with serum-free medium without phenol red starved SD rats osteoblasts at 2h,then give five groups concentrations Eucommia extract（0mg/ml,1mg/ml,10 mg/ml,50 mg/ml,100 mg/ml） interfere in SD rat osteoblasts and cells were incubated for 48 hours,set the control group（0mg/ml）,then MTT method detect cell proliferation; ibid steps,Western blot detect the expression levels of osteocalcin. Results After alkaline phosphatase staining,SD rats osteoblasts was purple,MTT method showed that the leaf extract on the proliferation of osteoblasts have a role in promoting（P〈0. 05） Compared with the control group,proliferation of osteoblas tsassociated with the drug concentration,wherein the most significant concentration was 50 mg/ml（P〈0. 01）. Western bolt results are shown that each dosing groups of osteocalcin protein levels were significantly elevated（P〈0. 05） compared with the control group,and also has a consistent concentration-dependent. Conclusion Eucommia extract can promote osteoblast proliferation through affecting the expression of osteocalcin and osteoblast proliferation has a concentration-dependent nature.

关 键 词：杜仲叶 成骨细胞 骨钙素 增殖 

分 类 号：R285.5[医药卫生—中药学]