槲皮素活化mtor信号通路对l929成纤维细胞 - 第三军医大学学报.docVIP

槲皮素活化mTOR信号通路对L929成纤维细胞增殖的影响 董海良1 ，张 颖1 ，黄谢梅1，黄 宏2 ，郭 韡2 ，邢 伟2 ，郝 进1 ，徐 祥2 (400010重庆，重庆医科大学附属第二医院皮肤科1； 400042重庆，第三军医大学第三军医大学大坪医院野战外科研究所第一研究室创伤、烧伤与复合伤国家重点实验室[摘要] 目的：探讨槲皮素（quercetin）对L929成纤维细胞增殖的影响及其分子机制。方法：采用MTT法检测槲皮素对L929细胞增殖的影响，荧光素酶报告基因法检测槲皮素对蛋白质翻译调控的影响，Western blot法检测槲皮素对mTOR 介导的蛋白质翻译调控通路关键基因mTOR的表达和4E-BP1的磷酸化,细胞周期调控蛋白cyclin D1的表达的影响及mTOR通路阻断mTOR、cyclin D1表达和4E-BP1的磷酸化明显增加。100ng/ml的雷帕霉素作用36h后，与对照组相比，槲皮素组P-4E-BP1、mTOR、cyclin D1蛋白表达明显增加，雷帕霉素组明显减少，雷帕霉素+槲皮素组蛋白表达明显增加，但较槲皮素组有所减少。 结论：槲皮素可通过诱导mTOR信号通路活化，进而促进细胞周期调控蛋白cyclin D1的表达，诱导L929细胞的增殖。 [关键词] 槲皮素；L929成纤维细胞；mTOR信号通路；细胞增殖 The effect of quercetin on mice fibrob-last cell line L929 proliferation through activating mTOR signal pathway Dong Hai-liang Zhang Ying Huang Xie-mei Huang Hong Guo Wei Xing Wei Hao jin xu xiang (Department of dermatology，The second Affiliated Hospital，Chongqing Medical University[Abstract] Objective：To determine the effect of quercetin on mice fibrob-last cell line L929 proliferation and explore it’s molecular mechanism．Methods：The effect of mice fibctrob-last cell line L929 proliferation was measured by MTT assay,the effect of quercetin on protein regulation was detected by luciferase reporter assay, the effect of quercetin on Expression of mTOR、cyclin D1 and p-4E-BP1,key genes in the mTOR-mediated protein translation pathway, were determined by Western Blot analysis．Results：l0-4mol/L )at the point of 36 hours,Quercetin significantly induced proliferation of mice fibrob-last cell line L929;The Luciferase reporter gene detection system showed:Compared with the control group, the RLU (relative light units) of 2.5×l0-4mol/L quercetin value increased significantly at the point of 36 hours;And quercetin increased the expression of mTOR and cyclin D1,and phosphorylation of 4E-BP1 in mice fibrob-last cell line L929 by western blot,and mTOR pathway blocking experiments .The role of 100ng/ml rapamycin, after 36h, compared with the control group, The P-4E-BP1, mTOR, cyclin D1 protein express