护心康片通过VEGF/VEGFR-2信号通路影响ApoE-/-小鼠动脉粥样硬化斑块的实验研究

Effect of Huxinkang Tablets on atherosclerotic plaque in ApoE-/-mice based on VEGF/VEGFR-2 signaling pathway

目的观察护心康片对动脉粥样硬化(AS)小鼠主动脉斑块的影响,探讨其作用机制。方法以高脂饲料喂养ApoE-/-小鼠12周建立AS小鼠模型,设空白组、模型组、护心康低剂量组、护心康中剂量组、护心康高剂量组和阿托伐他汀钙组,连续给药12周,分别行HE染色观察主动脉形态学改变,油红O染色观察脂质含量,免疫组化法测定CD34、MOMA-2、α平滑肌肌动蛋白(α-SMA)表达情况,RT-qPCR检测VEGF、VEGFR-2mRNA表达水平,Western blotting检测血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR-2)蛋白表达水平。结果与空白组比较,模型组小鼠主动脉有较大斑块形成,脂质含量多(P<0.01),α-SMA表达减少(P<0.01),CD34、MOMA-2表达增加(P<0.01),VEGF、VEGFR-2 mRNA及蛋白表达增加(P<0.01或P<0.05);与模型组比较,给药各组小鼠主动脉斑块均明显减小,脂质含量减少(P<0.01或P<0.05),α-SMA表达增加(P<0.01),CD34、MOMA-2表达减少(P<0.01),VEGF、VEGFR-2 mRNA及蛋白表达减少(P<0.01或P<0.05),其中护心康高剂量组效果最优且有优于阿托伐他汀钙组的趋势。结论护心康片可通过抑制AS小鼠主动脉斑块内血管新生来稳定斑块延缓AS,其机制可能与抑制VEGF/VEGFR-2信号通路有关。

Objective:To study the effects of Huxinkang Tablets on aortic plaque in mice with atherosclerosis,and to explore the possible mechanism of anti-atherosclerosis by Huxinkang tablets.Methods:ApoE-/-mice were given high-fat feed for 12 weeks to replicate AS mouse models.They were divided into the blank group,mod-el group,Huxinkang low-dose,middle-dose,high-dose groups and Atorvastatin calcium group.After 12 weeks of continuous administration,their aortic morphological changes were observed by HE staining,lipid contents were measured by oil red O staining,expressions of CD34,MOMA-2 andα-SMA were detected by immunohistochemis-try,expressions of VEGF and VEGFR-2 mRNA were detected by RT-qPCR,and expressions of VEGF and VEG-FR-2 protein were detected by Western blotting.Results:Compared with the blank group,mice in the model group had large plaque formation,whose lipid contents were high(P<0.01),α-SMA expressions were decreased(P<0.01),expressions of CD34 and MOMA-2 were increased,expressions of VEGF,VEGFR-2 mRNA and pro-tein were increased(P<0.01 or P<0.05);compared with the Model group,in all administered groups,aortic plaques were significantly reduced,lipid contents were reduced,α-SMA expressions were increased,expressions of CD34 and MOMA-2 were decreased,expressions of VEGF,VEGFR-2 mRNA and protein were decreased(P<0.01 or P<0.05),among which Huxinkang high-dose group had the best effect and tended to be superior to the Atorvastatin calcium group.Conclusion:Huxinkang Tablets can restrain angiogenesis in aortic plaques of mice with atherosclerosis to stabilize plaques and delay AS progression,and its mechanism may be related to the re-straint of VEGF/VEGFR-2 signaling pathway.