通痹灵合剂抗新生血管形成机制的研究

Study on inhibition mechanism of Tongbiling Composition on neovascularization

目的:研究通痹灵合剂抗新生血管形成的作用机制。方法:先随机分为造模组和正常对照组,采用弗氏完全佐剂于Wistar大鼠尾根部注射,足肿后随机分为造模对照组、消炎痛组、通痹灵合剂组。鸡胚尿囊膜模型分组:建立鸡胚尿囊膜模型(CAM),分为空白对照组、模型组,其中模型组加入20μl的10ng/L的血管内皮生长因子(VEGF)在载体上,再随机分为5组,加入各组大鼠血清或生理盐水20μl,分组为:VEGF+正常对照组,VEGF+造模对照组,VEGF+通痹灵合剂组,VEGF+消炎痈组,VEGF+1%生理盐水(VEGF组)。观察各组大鼠血清对CAM中一、二级新生血管的作用情况;另取各组大鼠右前和左后爪骨,运用RT-PCR方法检测各组关节组织的VEGF和VEGFR-1mRNA表达的影响。结果:各组CAM模型在10ng/L的VEGF刺激作用下,血管计数明显增加,VEGF+造模对照组血清作用于CAM后血管数目显著升高,与空白组比较差异明显(P<0.01)。VEGF+通痹灵合剂组血清的CAM血管计数中一、二级血管计数明显低于VEGF+造模对照组(P<0.01),消炎痛组CAM血管一、二级血管计数与造模对照组比较亦有明显降低(P<0.01)。RT-PCR图像显示出正常对照组、通痹灵合剂组、造模对照组及消炎痛组的大鼠关节组织VEGFmRNA表达,其中正常对照组及通痹灵合剂组明显弱于造模对照组与消炎痛组。正常对照组的VEGF-1mRNA图像不明显,通痹灵合剂组的VEGFR-1mRNA表达明显弱于造模对照组及消炎痛组。结论:本实验研究论证了通痹灵合剂通过抑制新生血管形成中VEGF产生及受体VEGFR-1在内皮细胞内的信号传递而阻断新生血管形成。

Objective： The study was to reveal the inhibition of neovascularization by Tongbiling. Methods： Rat model were established by injecting CFA at the root of the tail and divided into model control group, Indomethacin group and Tongbiling group at random. The serum and paws were collected after seven days treatment. 10ng/L VEGF solution was added I into CAM to stimulate the neovascularization, and to observe the influence of each group of serum on the first and second order new vessels. RT-PCR method was taken to determine the expression of mRNA of VEGF and VEGFR-1 of paws in different groups. Results： The number of vessels in the model control group was significantly more than that in the Indomethacin group and the Tongbiling group （P〈0.05）. The mRNA expression of VEGF and VEGFR-1 in Tongbiling group was dimmer than those in the model and the Indomethacin groups. Conclusion： Tongbiling could resist neovascularization by inhibiting the generation of VEGF and the signal transmission of VEGFR-1 in endothelial cell.