The Effect Of Huayu Qutan Recipe In Regulating HIF-1?/VEGF/VEGFR-2 Pathway On Aortic Plaque In AS Rabbits And Its Mechanism

Purpose:To observe the effect of Huayu Qutan recipe on regulating the expression of HIF-1?/VEGF/ VEGFR-2 pathway on the aortic lipid plaque in As rabbits and to explore the mechanism of the Huayu Qutan recipe on the atherosclerotic plaque.Material and method:Sixtyhealthy common grade male New Zealand rabbits were randomly divided into the blank control group,the model group,the Huayu Qutan Recipe group and the Simvastatin group.Atherosclerotic rabbit model wasestablished by immune-induced endothelial injury and by feeding with high-fatdiet.The control group was given normal feed.The high-fat group was given high-fat food.After 8 weeks,each group was given the corresponding drug intervention;the control group and the model group were given the same volume of saline,1times a day for 4 weeks before taking materials.The levels oftriglyceride(TG)total cholesterol(TC)TC,low-density lipoprotein cholesterol(LDL-C)andhigh-density lipoproteincholesterol(HDL-C)in serumwere detected by the automatic biochemical analyzer.HE staining and the oil red O staining were used to observe the pathological changes of the aorta in rabbits.JEOR 801 D morphological image analysis system software was used to measure the thickness of aorta intima and media.Immunohistochemical method was adopted to determine CD31 mean optical density value.The contents ofhypoxia inducible factor(HIF)?vascular endothelial growth factor(VEGF)?Vascular Endothelial Growth Factor Receptor 2(VEGFR-2)in serum were detected by ELISA.Western blot was used to detect the expression of HIF-1?,VEGF and VEGFR-2 in the aorta.Results:1.The results of serum lipid content of rabbits in each group showed: Compared with the blank control group,the levels of TG,TC and LDL-C in the model group were significantly increased(P<0.01)and HDL-C levels were significantly decreased(P <0.01).Compared with the model group,the levels of TG,TC and LDL-C in the Huayu Qutan Recipe group and the Simvastatin group were significantly decreased(P <0.05 or P <0.01),and HDL-C content was significantly increased(P <0.05 or P <0.01).2.The results of HE staining and the oil red O staining in each group showed : in the blank control group,the intima of the aortic wall was smooth and complete,with uniform thickness.The endothelial cells were arranged in a clear and orderly manner,without lipid deposition.The smooth muscle cells were arranged in a regular manner,and no pathological changes were observed.In the model group,the intima cells of the aortic duct wall proliferated,and the intima were significantly thickened and rough and not smooth.The vascular wall was thickened,with a large number of foam cells and lipid deposition,and the smooth muscle cells were irregularly arranged.In the Huayu Qutan Recipe group,the intima of the aortic wall was smooth and complete,with a small amount of foam cells and lipid deposition.The thickening degree of the aortic wall was significantly reduced,and smooth muscle cells were arranged regularly.In the Simvastatin group,the intima of the aortic tube wall was smooth and complete,with a small amount of foam cells and lipid deposition.The thickness of the tube wall was significantly reduced,and smooth muscle cells were more regular.Oil red O staining showed that the intima of the aortic wall of the blank control group was smooth,thin,soft,shiny,white,without plaques or bumps.In the model group,the intima of the aortic wall was rough and unsmooth,thickened,and there were a large number of red-stained and variously sized uplift plaques combined into a lumen with convexity,and the elasticity of the wall decreased.In the Huayu Qutan Recipe group,The intima of the aortic wall is basically smooth,lipid striations and plaques are scattered or thin strips with occasional uplift,and the wall has good elasticity.In the Simvastatin group,smooth vascular intima,lipid striations and plaques were observed in the aortic wall,with occasional uplift and good elasticity.3.The results of quantitative analysis of aorta histopathology and morphology in each group showed: Compared with the blank control group,Compared with the blank control group,the thickness of intimaand tunica mediain the model group were significantly increased(P<0.01).Compared with the model group,The thickness of intima and tunica media in the Huayu Qutan Recipe group and the Simvastatin group were significantly decreased(P <0.01).4.The results of immunohistochemical in each group showed: the blank control group,there was a small amount of positive CD31 expression in the intima of aortic wall,and the expression product was brown-yellow particles.In the the model group,he intima of the aorta was obviously thickened,and there were continuous or discontinuous small lumen with brown color near the inner elastic plate,which was the new small blood vessel and showed strong positive expression.In the Huayu Qutan Recipe group,A small number of brownyellow granules were observed in the intima of aorta,and the expression of CD31 was significantly decreased.In the Simvastatin group,A small number of brown-yellow granules were observed in the intima of aorta,and the expression of CD31 was significantly decreased.The result of CD31 mean optical density value in each group showed: compared with the blank control group,the CD31 mean optical density value in the model group were significa-ntly increased(P<0.01).Compared with the model group,the CD31 mean optical density value in the Huayu Qutan Recipe group and the Simvastatin group were significantly decreased(P<0.01).5.The result of serum HIF-1? and VEGF levels in each group showed: compared with the blank control group,the contents of HIF-1?,VEGF in the model group were significantly increased(P<0.01).Compared with the model group,the levels of HIF-1?,VEGF in the Huayu Qutan Recipe group and the Simvastatin group were significantly decreased(P <0.05 or P <0.01).6.The expression levels of HIF-1?,VEGF and VEGFR-2 in the aorta showed: compared with the blank control group,The expressions of HIF-1?,VEGF and VEGFR-2 in the aorta of the model group were significantly increased(P <0.01).Compared with the model group,the expression of HIF-1?,VEGF and VEGFR-2 in the Huatan Qutan Recipe and the Simvastatin were significantly decreased(P<0.01).Conclusion:Huayu Qutan Recipe can significantly decreased the levels of HIF-1? and VEGF in the serum and the expression of angiogenic factors in aortic lipid plaques of AS rabbits.It has been proved that this method can inhibit angiogenesis in the plaques,Thus,it can delay the occurrence and development of AS and play an anti-asrole.