NaF与H_2O_2对大鼠真皮成纤维细胞血管内皮生长因子表达的影响  

Effects of NaF and H_2O_2 on expression of vascular endothelial growth factor in fibroblasts of rats

作　　者：吴昊昱[1] 吴璇[1] 魏振宏[1] 刘朋飞[1] 董超[1] 冯业童[1] 刘迪[1] 石毅[1] 孙波[1,2] 

机构地区：[1]吉林大学药学院生物工程试验中心,吉林长春130021 [2]吉林大学药学院实验药理与毒理学教研室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2012年第5期948-951,F0003,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省卫生厅科研基金资助课题(2009Z043);吉林大学博士研究生交叉学科科研基金资助课题(2011J023)

摘　　要：目的:比较不同剂量的氟化钠(NaF)与过氧化氢(H2O2)对成纤维细胞表达血管内皮生长因子(VEGF)的影响,探索促进VEGF表达的简单有效的方法,为优化组织工程真皮培养条件提供理论参考。方法:酶消化法分离大鼠皮肤成纤维细胞,以不同剂量NaF(0.05和0.50 mmol.L-1)、H2O2(10 nmol.L-1和10μmol.L-1)以及两者联合(NaF 0.05mmol.L-1,H2O210μmol.L-1)分别作用于成纤维细胞,在第0、24及48h采用CCK-8法测定细胞的增殖活性,在第48小时采用ELISA法和免疫组织化学法测定其对VEGF表达的影响。结果:不同剂量NaF、H2O2以及两者联合作用对成纤维细胞增殖无抑制作用,与空白对照组比较差异无统计学意义(P>0.05);NaF实验组和H2O2实验组的VEGF表达量均明显提高,与空白对照组比较差异有统计学意义(P<0.05),而NaF与H2O2两者联合作用效果最为明显,可提高40%(P<0.05)。结论:NaF与H2O2联合作用能在不影响成纤维细胞正常增殖的前提下,有效地促进VEGF的表达。Objective To compare the effects of different doses of sodium fluoride（NaF） and hydrogen peroxide（H2O2） on expression of vascular endothelial growth factor（VEGF） in fibroblasts of rats,and to explore a simple and effective way to promote the expression of VEGF,and in order to provide some theoretical reference to optimize the dermis culture condition in tissue engineering.Methods The rat fibroblasts were isolated by enzyme digestion,and they were cultivated respectively with DMEM medium contained different doses of sodium fluoride（0.05 and 0.50 mmol·L-1）and H2O2（10 nmol·L-1and 10 μmol·L-1）,or with both two factors（NaF 0.05 mmol·L-1,H2O2 10 μmol·L-1）.The proliferation ability was evaluated by CCK-8 assay at 0,24 and 48 h after the fibroblasts were treated with these factors.And the effects of NaF and H2O2 on the expression of VEGF were evaluated by ELISA assay and immunohistochemical method at 48 h.Results There was no evidently inhibitory effects of different doses of NaF and H2O2 used alone or combined on the proliferation of fibroblas,there was no significant difference compared with blank control（P0.05）.The expressions of VEGF in both NaF group and H2O2 group were greatly improved,there was significant difference compared with blank control group（P0.05）.However,the expression of VEGF in the fibroblasts treated with combined two factors was most enhanced,which could increase 40%（P0.05）.Conclusion The combination of NaF and H2O2 can keep the normal proliferation activity of fibroblasts,and promote the expression of VEGF effectively.

关 键 词：成纤维细胞 血管内皮生长因子 氟 过氧化氢 

分 类 号：R318.08[医药卫生—生物医学工程]