单核巨噬细胞集落刺激因子诱导创伤后成纤维细胞生成血管内皮生长因子的研究  

GMCSF induces production of VEGF via PKC-NF-κB signal pathway in human fibroblasts during wound healing

作　　者：李晓光[1] 方勇[1] 姚敏[1] 俞为荣[1] 徐鹏[1] 郭兵[1] 毛志刚[1] 杨鹏高[1] 

机构地区：[1]上海交通大学医学院附属第三人民医院颌面外科,201900

出　　处：《中华医学美学美容杂志》2015年第4期229-233,共5页Chinese Journal of Medical Aesthetics and Cosmetology

基　　金：上海市教委优秀青年基金资助项目（zzjdyx12112）;金赛烧伤外科医师科研基金资助项目（201210）

摘　　要：目的 探讨创伤愈合过程中单核巨噬细胞集落刺激因子（GMCSF）调控新生血管化的分子机制.方法 将成纤维细胞分为空白对照组、GMCSF组、GMCSF＋ Go6976组以及Go6976组,空白组应用GMCSF溶解液PBS处理;GMCSF组应用50 ng/L的GMCSF处理;GMCSF＋Go6976组先加入50 ng/L的GMCSF和10μmol/L的Go6976处理;Go6976组用10 μmol/L的Go6976处理.应用GMCSF作用成纤维细胞后,采用Western印迹法观察蛋白激酶磷酸化水平;采用酶联免疫吸附试验检测血管内皮生长因子（VEGF）表达,采用凝胶电泳迁移试验检测核因子-κB的活化.结果 GMCSF作用成纤维细胞45 min蛋白激酶C蛋白相对表达量0.434±0.056,与0 min 0.224±0.035比较,差异有统计学意义（P＜0.05）;ELISA显示,GMCSF＋ Go6976处理组VEGF表达量为39.26土7.63,与GMCSF组的86.75±10.78比较,差异有统计学意义（P＜0.05）;凝胶电泳迁移实验结果显示,GMCSF组核因子-κB为82835±4565,GMCSF＋Go6976组为71279±3658,差异有统计学意义（P＜0.05）.结论 GMCSF可通过PKC活化核因子-κB,从而诱导人皮肤成纤维细胞VEGF的表达.Objective To explore the molecular mechanisms for GMCSF regulating angiogenesis during wound healing.Methods Human fibroblasts were divided into control group,GMCSF group,GMCSF＋Go6976 group and Go6976 group.The control group was treated with GMCSF solution PBS,the GMCSF group treated with 50 ng/L GMCSF,the GMCSF＋ Go6976 group treated with 50 ng/L GMCSF and 10 μmol/L Go6976,and the Go6976 group treated with 10 μmol/L Go6976.The total VEGF proteins were extracted from the fibroblasts treated with GMCSF.Western blotting was employed to determine levels of PKC.The fibroblasts were treated by CMCSF.VEGF protein was analyzed by ELISA;NF-κB activation was analyzed by EMSA.Results The relative expression level of reached PKC 0.434±0.056 to the peak at 45 min in the fibroblasts affected by GMCSF,com-pared with that of PKC （0.224 ±0.035）,with significant difference （P〈0.05）.Compared with the GMCSF group （VEGF protein expression 86.75± 10.78）,the GMCSF＋Go6976 group （VEGF protein expression 39.26±7.63） showed that the expression level of VEGF significantly inhibited （P〈0.05） in ELISA.In EMSA,GMCSF group of nuclear factor kappa B was 82835 ±4565;and GMCSF＋group （71279 ± 3658） Go6976 binding activity of nuclear factor kappa B and DNA was significantly lower than that in the GMCSF group （P〈0.05）.Conclusions GMCSF up-regulates production of VEGF through activating PKC-NF-κB signal pathway in human fibroblasts.

关 键 词：单核巨噬细胞集落刺激因子 血管内皮生长因子 血管新生 

分 类 号：R641[医药卫生—外科学]