杜仲叶通过激活ERK及AKT磷酸化促进大鼠成骨细胞增殖的研究  

Eucommia leaf promotes rat osteoblast proliferation by activating the phosphorylation of ERK and AKT

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

机构地区：[1]南昌大学第二附属医院,南昌330006 [2]江西中医学院

出　　处：《中国骨质疏松杂志》2013年第3期217-220,共4页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金(81160508);江西省自然科学基金(GZY0170)

摘　　要：目的研究杜仲叶提取物对大鼠成骨细胞的增殖作用及其分子机制。方法选取新生SD大鼠的乳鼠颅骨通过消化法分离出乳鼠成骨细胞,并用碱性磷酸酶染色进行细胞鉴定;先用杜仲叶提取物分别以0 mg/ml,5 mg/ml,20 mg/ml,40 mg/ml,60 mg/ml五种不同浓度梯度对大鼠成骨细胞干预,2d后用MTT方法检测细胞增殖情况;用无血清无酚红的培养基饥饿大鼠成骨细胞2 h后,分别以0mg/ml,5 mg/ml,20 mg/ml,40 mg/ml,60 mg/ml五种不同浓度杜仲叶提取物干预大鼠成骨细胞,2 h后Western blot检测ERK和AKT的活化情况。结果碱性磷酸酶染色后的成骨细胞呈紫红色(特异性染色),MTT法结果显示杜仲叶提取物促进大鼠成骨细胞的增殖,并具有浓度依赖性;Western blot结果显示杜仲叶提取物可使ERK及AKT磷酸化水平提高,并具有浓度依赖性。结论杜仲叶提取物通过ERK通路及AKT通路促进大鼠成骨细胞的增殖。Objective To investigate the effect of proliferation and the molecular mechanism. Methods the extraction of eucommia leaves on rat osteoblast calvaria of new-born SD rats. The cells were identified The osteoblasts were isolated and cultured from the with alkaline phosphatase staining. Rat osteoblasts were treated with 5 different concentration gradients of leaf extraction： 0 mg/ml, 5 mg/ml, 20 mg/ml, 60 mg/ml, and 100 mg/ml. Cell proliferation was detected using MTT method 2 days later. After the addition of serum-free and phenol red-free medium for 2 h, 0 mg/ml, 5 mg/ml, 20 mg/ml, 60 mg/ml, or 100 mg/ ml leaf extraction was additioned to the medium of the osteoblast culture. Then the expression of ERK and AKT was detected using Western blotting 2 hours later. Results After alkaline phosphatase staining, osteoblasts were stained in purple （specific staining）. The results of MTT methods showed that leaf extraction promoted rat osteoblast proliferation in a dose-dependent manner. The results of Western blotting showed that leaf extraction could improve the phosphorylation levels of ERK and AKT in a dose - dependent manner. Conclusion The extraction of eucommia leaves promotes rat osteoblast proliferation through ERK and AKT pathways.

关 键 词：杜仲叶 成骨细胞 ERK AKT 增殖 

分 类 号：R285.5[医药卫生—中药学]