杜仲叶提取物槲皮素通过激活ERK磷酸化促进BMSCs增殖的研究  

A Study of Quercetin Extracted from Eucommia Leaf Promoting the Proliferation of Bone Marrow Derived Mesenchymal Stem Cells through the Phosphorylation of ERK

作　　者：陈林攀[1] 邓鸣涛[1] 杜川[1] 方宁[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]江西中医药大学,江西南昌330006

出　　处：《时珍国医国药》2014年第12期2845-2847,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81160508)

摘　　要：目的探讨杜仲叶提取物槲皮素对SD大鼠骨髓间充质干细胞增殖的影响及其分子机制。方法采用经典的全骨髓培养法培养MSC,通过倒置相差显微镜观察细胞形态特征、流式细胞术分析其表面标志的方法鉴定BMSC。取第三代BMSC为实验材料,设用0.5,1,2,4μg·ml-1的杜仲叶提取物槲皮素处理细胞的为实验组,未加槲皮素为空白对照组,采用MTT法检测MSC增殖水平。用无血清无酚红培养基饥饿大鼠MSC 6h后,分别以终浓度0,0.5,1,2,4μg·ml-1的杜仲叶提取物槲皮素干预细胞15min,Western blot检测ERK磷酸化水平的变化。结果第3代BMSC呈现CD90、CD29阳性,CD45阴性特征。MTT结果显示杜仲叶提取物槲皮素均能促进BMSC增殖。Western blot结果显示杜仲叶提取物槲皮素促使ERK的磷酸化水平升高。结论杜仲叶提取物槲皮素通过激活ERK磷酸化促进BMSC增殖。Objective To explore the effect of quercetin extracted from Eucommia leaf on mesenchym stem cell （MSC） prolifera- tion and its possible signal transduction mechanism. Methods BMSC culture was performed with the adhering culture method, u- sing the phase contrast microscope to observe cell morphology and flow cytometry to detect the cell surface antigen to identify the BMSC. The third generation of BMSC was used as experimental material. The experimental group was designed with 0. 5 ,1,2 ,4μg·ml^-1 of quercetin extracted from Eucommia leaf. The group without quercetin was set as the control group. By using the MTY method to detect the proliferation of BMSC. Adding the end concentration 0,0.5,1,2,4μg·ml^-1 of quercetin extracted from Eu- commia leaf to the BMSC for 15min after starving the BMSC for 6h by the serum - free and phenol red - free medium. Then we de- termine the changes of the phosphorylation of the ERK level using the Western blot. Results Cell surface antigen detection is CD90（ ＋ ） ,CD29（ ＋ ） and CD45 （ - ）. MTT results showed that the quercetin extracted from Eucommia leaf can promote BMSC prolieferation . Western blot results proved this drug can stimulate the phosphorylation of the ERK. Conclusion The quercetin ex- tracted from Eucommia leaf Promote the proliferation of bone marrow derived mesenchymal stem cells through the phosphorylation of the ERK pathway.

关 键 词：杜仲叶 槲皮素 骨髓间充质干细胞 ERK 增殖 

分 类 号：R285.5[医药卫生—中药学]