基于bFGF基因表达探讨缺氧条件下补阳还五汤促进体外血管三维模型的血管新生分子机制

Study on Molecular Mechanism of Neovascularization of the Three-Dimensional Model of Blood Vessels Which Promoted by Buyang Huanwu Decoction in Vitro under Hypoxia Condition Based on bFGF Gene Expression

目的:基于bFGF基因表达情况调控缺氧条件下补阳还五汤促进体外血管三维培养模型中新生血管的分子机制。方法:于Matrigel基质胶上种植腹主动脉血管段构建体外血管三维培养模型,常氧条件下培养5 d后,给予24 h缺氧干预,平行进行补阳还五汤和辛伐他汀治疗,利用倒置显微镜观察新生血管的生长情况,利用实时定量PCR技术分析bFGF、PDGFB、Ang1的基因表达水平。结果:倒置显微镜观察各实验组均有新生血管形成,其中模型组生长最为旺盛,但杂乱无序,补阳还五汤治疗后,新生血管规则有序。RT-PCR结果显示,与正常组相比,模型组bFGF基因表达明显上调(P <0. 01)、PDGFB基因表达上调(P <0. 05),Ang1基因表达下调(P <0. 05);基于补阳还五汤治疗后,bFGF基因表达上调(P <0. 05)、PDGFB基因表达明显上调(P <0. 01),Ang1基因表达上调(P <0. 05)。结论:缺氧条件下,经补阳还五汤治疗后,上调体外血管三维模型的新生血管相关基因水平表达,进而促进新生血管形成。

Objective:The molecular mechanism of neovascularization of the three-dimesional model which was promoted by Buyang huanwu Decoction under hypoxia condition was regulated based on the expression of bFGF gene.Methods:The three-dimesional model of vascularization was established by implanting abdominal aorta in Matrigel matrix.After incubation under normoxia for 5 days,the model was given hypoxia intervention for 24 hours and parallel to the treatment of Buyang Huanwu Decoction and simvastatin.The growth of neovascularization was observed by inverted microscope.The expressions of b FGF,PDGFB and Ang1 genes were analyzed by real time quantitative PCR.Results:There was neovascularizition which was observed by inverted microscope in each experimental group.The neovascularizition had the most vigorous growth in the model group,but disorderly.After the treatment of Buyang Huanwu Decoction,the neovascularization grew orderly.The result of RT-PCR showed that the expressions of bFGF(P<0.01)and PDGFB(P<0.05)genes were higher and the expression of Ang1 gene was lower(P<0.05)in the model group compared with those of the normal group.After the treatment of Buyang Huanwu Decoction,the expressions of bFGF(P<0.05),PDGFB(P<0.05)and Ang1(P<0.05)genes were higher compared with those of the model group.Conclusion:After the treatment of Buyang Huanwu Decoction,the expressions of several genes were high in neovascularization of the three-dimensional model of blood vessels in vitro under hypoxia condition,and thus promoted the formation of neovascularization.