杜仲叶提取物槲皮素通过激活ERK磷酸化促进BMSC增殖的研究-外科学(骨外)专业论文.docxVIP

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 其可能的分子作用机制。 方法：采用经典的全骨髓培养法培养 BMSC，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浓度梯度处理细胞的 为实验组,未加槲皮素为空白对照组，MTT 法检测 BMSC 的增殖。饥饿大鼠 MSC 6h 后，分别以五种不同的终浓度 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 活化程度。 结果：对培养的细胞的形态进行观察，结果符合 SD 大鼠 BMSC 干细胞的 特征；第三代 BMSC 的 CD90CD29 阳性，CD45 阴性。MTT 结果显示，槲皮素 能促进 BMSC 增殖；Western blot 结果显示，槲皮素促使 ERK 的磷酸化水平升 高，两者均有浓度依赖性。 结论：杜仲叶提取物槲皮素可能经 ERK 活化途径促 BMSC 增殖。 关键词：杜仲叶；槲皮素；骨髓间充质干细胞；ERK；增殖 II ABSTRACT Objective：To explore the effect of quercetin extracted from eucommia leaf on mesenchym stem cell (MSC) proliferation and its possible signal transduction mechanism. Methods：BMSC culture was performed with the adhering culture method.Using the phase contrast microscope to observe cell morohology 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 with0.5、1、 2 、 4μg·mL-1 of quercetin extracted from eucommia leaf.The group without quercetin was set as the control group.By using the MTT method to detect the proliferation of BMSC. Adding the end concentration 0,0.5,1,2,4μg·mL-1 of quercetin extracted from eucommia leaf to the BMSC for 15min after starving the BMSC for 6h by the serum-free and phenol red-free medium.Then we detemine 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 in a concentration dependent manner.Western blot results proved this drug can stimulate the Phosphorylation of the ERK . Conclusion：The Quercetin extracted from eucommia leaf Promote the prolifera -tion of bone marrow derived mesenchymal stem cells through the Phosphorylation of the ERK pathway. Key Words: Eucommia leaf; Quercetin; BMSC; ERK; proliferatio