摘要
目的研究凉血退赤方(LXTCF)含药血清对人脐静脉内皮细胞(ECV304)血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(b-FGF)、促血管生成素1(ANG-1)及色素上皮衍生因子(PEDF)基因及蛋白表达的影响,探讨其抑制角膜血管新生的可能作用机制。方法制备LXTCF含药血清,以不同浓度的LXTCF含药血清(10%、20%)作用于ECV304,以正常大鼠血清(10%、20%)处理的ECV304为空白对照组,以单纯培养体系处理的ECV304为正常对照组。采用Western Blot、实时荧光定量PCR(RT-PCR)检测ECV304细胞VEGF、b-FGF、ANG-1及PEDF蛋白及基因表达的变化。结果与空白对照组(20%)[(0.695±0.028),(1.28±0.08)]比较,20%LXTCF含药血清可明显降低ECV304细胞VEGF[(0.416±0.053),(0.56±0.13)]、b-FGF[(0.306±0.053),(0.64±0.11)]蛋白及基因表达(P<0.05);含药血清(10%、20%)对ANG-1及PEDF蛋白及基因表达均无明显影响(P>0.05)。结论下调VEGF和b-FGF基因及蛋白表达,从而抑制角膜血管新生,可能是LXTCF治疗CRNV性疾病的机制之一。
Objective： To study the influence of drugcontaining serum of Lian Xue Ttui Chi Fang （LXTCF） on VEGF, b- FGF, ANG - 1 and PEDF of human umbilical vein endothelial ceils and explore its possible mechanism. Methods ： The drug - containing serum of LXTCF was prepared with different concentrations of LXTCF, containing the serum （ 10%, 20% ） to affect on ECV304, and taking ECV304 normal serum （ 10%, 20% ） as a blank control group, and treating ECV304 of pure culture system as a normal control group. Western Blot was used to make real - time quantitative PCR （ RT - PCR） and to detect the changes of ECV304 cell, VEGF, b - FGF, ANG- 1 and PEDF protein and gene expression. Results： Compared with the control group （20%） [ （0. 695 ± 0. 028 ）, （1.28 ± 0.08） ], 20% of LXTCF containing serum can significantly reduced the ECV304 cell VEGF [ （0. 416±0. 053）, （0. 56 ±0. 13） ], b - FGF [ （0. 306 ±0. 053 ）, （0. 64 ± 0. 11 ） ] gene and protein expression （ P 〈 O. 05 ）. The drug - containing serum （ 10%, 20% ） had no significant difference （P 〉 0. 05 ） on ANG - 1 and PEDF protein and gene expression. Conclusion ： The lowering of VEGF and b - FGF gene and protein expression can inhibit corneal neovascularization, which may be one of CRNV disease mechanisms of LXTCF treatment.
出处
《云南中医中药杂志》
2015年第2期61-64,共4页
Yunnan Journal of Traditional Chinese Medicine and Materia Medica
基金
云南省科技厅面上项目(NO:2011FZ257)
