杜仲叶通过Shp2激活Erk1/2促进BMSCs增殖的研究  

Duzhongye Activates Erk1 /2 Through Shp2 to Promote BMSCS Proliferation

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

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

出　　处：《时珍国医国药》2013年第6期1286-1288,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81160508);江西省自然科学基金(No.GZY0170)

摘　　要：目的研究杜仲叶提取物对大鼠BMSCs的增殖作用及其分子机制。方法使用密度梯度离心和贴壁培养法从SD大鼠股骨骨髓中获取BMSCs,通过流式细胞仪检测细胞表面抗原;分别用0,5,20,60,100 mg/ml的杜仲叶提取物和20mg/ml杜仲叶提取物+10μmol/L Shp2抑制剂NSC87887处理BMSCs,2 d后BrdU检测细胞增殖情况;用无血清无酚红培养基饥饿大鼠BMSCs 6 h后,一组分别加入0,5,20,60,100 mg/ml的杜仲叶提取物处理15 min,另一组用10μmol/LShp2抑制剂处理预处理2 h后加入20 mg/ml杜仲叶提取物处理15 min,Western blot检测p-Shp2(Tyr580)和Erk1/2磷酸化水平的变化。结果流式细胞仪细胞表面抗原检测显示CD29、CD90呈阳性,CD34、CD45呈阴性;BrdU结果显示杜仲叶提取物促进BMSCs增殖,并具有浓度依赖性,NSC87887可以抑制其促进作用;杜仲叶提取物可以诱导BMSCs的p-Shp2(Tyr580)和Erk1/2磷酸化水平升高,而对诱导Erk1/2磷酸化的促进作用是通过激活Shp2来完成的。结论杜仲叶提取物通过Shp2/Erk途径促进大鼠BMSCs增殖。Objective To study its molecular mechanism of Duzhongye extract on the proliferation of rat BMSCs.Methods Using density gradient centrifugation and adherent culture of BMSCs from the SD rat femur bone marrow;Cell surface antigen is detected by flow cytometry;0,5 mg/ml,20 mg/ml,60 mg/ml,100 mg/ml respectively Duzhongye extract and 20mg/ml Duzhongye extract ＋ 10 μM Shp2 inhibitor NSC87887 processing BMSCs the 2 days,then BrdU detection cell proliferation;Groups were added with serum-free medium without phenol red starved rats BMSCs at 6h,then by 0,5 mg/ml,20 mg/ml,60 mg/ml,100 mg/ml Duzhongye extract treatment 15min,another group pretreatment with 10μM Shp2 inhibitors two hours,then added 20 mg/ml Duzhongye extract processing 15 min,Western blot detection p-Shp2（Tyr580） and Erk1/2 phosphorylation levels.Results Cell surface antigen detection is CD29-positive,CD90-positive and CD34-negative,CD45-negative;BrdU results showed that Duzhongye extract promotes BMSCs proliferation in a dose-dependent manner and NSC87887 can inhibit its role in promoting;BMSCs can be induced p-Shp2（Tyr580） and Erk1/2 phosphorylation levels increased by Duzhongye extract,while the role of induced Erk1/2 phosphorylation is accomplished by activating Shp2.Conclusion The effect of Duzhongye extract can promote the proliferation of rat BMSCs by Shp2/Erk pathway.

关 键 词：杜仲叶 BMSCS Shp2 ERK1 2 增殖 

分 类 号：R285.5[医药卫生—中药学]