他汀类药物对血管样结构物生长的促进作用

Promoting effect of statin on the growth of vessel-like structure

目的观察不同他汀类药物对血管样结构物生长及对培养人脐静脉内皮细胞(HUVEC)血管内皮生长因子(VEGF)表达的影响,探讨体外实验中他汀类药物是否具有独立于血脂调节作用以外的促进血管新生作用。方法①多种他汀类药物血管新生共同培养:加入匹他伐他汀、阿托伐他汀、普伐他汀培养后,采用倒置显微镜计数血管样结构物管腔形成数,并与阳性对照组、血管新生抑制剂组比较。②匹他伐他汀血管新生共同培养:添加不同浓度的匹他伐他汀培养后,采用血管新生定量软件分析系统计算血管样结构物管腔面积、管腔长度、管腔交差点数和管腔支数,并与阳性对照组进行比较。③HUVEC培养:添加不同浓度的匹他伐他汀培养后回收HUVEC,采用Westernblot方法检测VEGF蛋白的表达。结果①多种他汀类药物培养:匹他伐他汀1×10-10mol/ml、阿托伐他汀1×10-6mol/ml,普伐他汀1×10-5mol/ml浓度时,血管样结构物管腔支数明显高于对照组和血管新生抑制剂组(P<0.05),与阳性对照组的差异无显著性(P>0.05)。②匹他伐他汀培养:匹他伐他汀1×10-11～1×10-10mol/ml浓度时,血管样结构物管腔面积明显增大,与阳性对照组的差异无显著性(P>0.05)。匹他伐他汀1×10-10mol/ml浓度时,血管样结构物管腔长度、管腔交差点数和管腔支数均明显增加(P<0.05),与阳性对照组的差异无显著性(P>0.05)。③Westernblot法检测结果:匹他伐他汀1×10-8～1×10-12mol/ml浓度时,HUVEC培养细胞VEGF蛋白表达增强。结论他汀类药物促进了血管样结构物的生长,匹他伐他汀能诱导人脐静脉内皮细胞培养对VEGF蛋白的表达,推测他汀类药物可能具有独立于血脂调节作用以外的促进血管新生作用,且该作用与药物剂量有关。

Objective To observe the influence of Statins on the growth of vessel-like structure and the cytokines＇ expression of cultivated human umbilical vessel endothelial cells （HUVEC）, and to test the pro-anglogenesis effect of Statins independent of regulation of blood lipids. Methods The HUVECs and fibroblasts were co-cultured with 1 × 10^-8 - 1 × 10 ^-10 mol/ml Pitavastatin, 1 ×10^-6 -1× 10^-8 mol/ml Atorvastatin or 1 × 10^-5 -1 ×10^-7 mol/ml Provastatin in multiple 6 pores Petri dish. And then the number of vessel-like structures in each pore was calculated and compared with negative control （DMSO） group, positive control （VEGF） group and the angiogenesis-antagonist group. Pitavastatin in a serial concentrations （1 × 10^-10 × 10^-13 mol/ml） was added into the cells co-culture system, and then the quantitative angiogenesis analysis software was used to calculate the luminal area of the vessel-like structure, the length of the lumen, the crossing-point of the vessels, the number of vessel branches and compared with the positive control group. After treatment with Pitavastatin, the HUVECs were collected and the expression of VEGF was assessed by western blot. Results Under the conditions of 1 × 10^-10 mol/ml Pitavastatin,1 ×10^-6 mol/ml Atorvastatin or 1 × 10^-5 mol/ml Provastatin, the vessel-like structures were more than that in the negative control and the angiogenesis-antagonist group, no significant difference was seen as compared with the positive control group. The luminal area of the vessel-like structure increased with the treatment of Pitavastatin in concentrations of 1 × 10^-11 × 10^-10 mol/ml, and the length of lumen, the crossing-point of the vessels and the number of vessel branches increased when 1 ×10^-10 mol/ml Pitavastatin was added, no significant difference was seen as compared with the positive control group. The results from western blot implied that the expression of VEGF in HUVECs augmented under circumstances of 1 ×10^-8 - 1 × 10^-13 mol/ml Pitavastatin. Conclusions Statins induce the growth of vessel-like structure and the expression of VEGF in cultivated HUVECs. Statins may have pro-angiogenesis effect with dose-dependent and independent of the effect of blood lipid regulation. （Shanghai Med J, 2005,28 ： 866-868）