Study On The Anti-atherosclerotic Effect Of Resveratrol And Its Effect On Smads Signaling Pathway

Ischemic cerebrovascular disease is the main pathological basis for atherosclerosis,in the process of formation of atherosclerosis,inflammatory reaction throughout the whole process of occurrence and development of atherosclerosis,Smads signal pathways can inhibit lipid metabolism,inhibit inflammation and oxidative stress play a role of resistance to atherosclerosis,resveratrol mainly through anti-inflammatory,inhibiting oxidative modification of low density lipoprotein,inhibition of foam cell formation,restrain plaque inside new blood vessels,such as antioxidant role of resistance to atherosclerosis.However,the relationship between resveratrol,Smads signaling pathway,lipid metabolism,inflammatory response,and atherosclerosis remains uncertain.This study to determine whether atherosclerosis resistant effect of resveratrol and its through Smads signal pathway of anti-inflammatory,resistance to atherosclerosis mechanism,on the basis of successful replication of the rabbit model of atherosclerosis,using different imaging techniques for the imaging characteristics of carotid atherosclerotic comparative analysis,to observe the effects of resveratrol on imaging change at the same time,Immunohistochemistry and Western-blot were used to study the Smads signaling pathway as well as the expression of macrophage markers RAM11,Actin,and vascular endothelial cell marker CD31 in rabbit carotid artery atherosclerosis,and to compare and analyze the levels of lipid and Lp-PLA2 in rabbit aortic arch atherosclerosis.The research of this paper includes the following contents:1.Establishment and evaluation of rabbit models of carotid and aortic arch atherosclerosisObjective: To establish a rabbit model of carotid artery and aortic arch atherosclerosis.Methods: Eight healthy common grade New Zealand white rabbits,male,weight2.7-3.3kg,age 3 months,4 in the carotid atherosclerosis group and 4 in the aortic arch atherosclerosis group;In the carotid atherosclerosis group,2 rabbits in the control group were given normal diet,high-fat diet combined with partial carotid artery ligation.Operation is as follows: the ear marginal vein indwelling trocar,ear marginal vein slowly push note 5% of phenobarbital sodium injection 100 mg/kg,anesthesia and surgery under aseptic conditions,neck midline incision,show and separation of the left carotid artery,in LJZ-4D under surgical microscope with a suture needle through the carotid artery middle position,10-0 silk ligation of artery block parts of blood vessels,causing about 50% stenosis,step by step a closing a wound.After carotid artery ligation,cervical blood vessel ultrasound examination was conducted 3months after feeding,and the carotid artery was dissected for HE staining.Aortic arch atherosclerosis group 4,normal diet group 2,high fat diet group 2.Raise for 3 months.Then anesthesia was performed,the aortic arch was dissected,and HE staining was performed.Results: The control group showed smooth carotid artery with no plaque,while the experimental group showed partial stenosis of carotid artery with plaque formation.Local gross anatomy showed that the cervical vascular surface of the control group was smooth and smooth,without plaque formation,while the ligation line of the experimental group showed local plaque formation.HE staining showed that the tube wall of the control group was smooth,with no intimal hyperplasia and normal morphology of smooth muscle layer without plaque formation.In the experimental group,the carotid intima was thickened,the smooth muscle layer had irregular morphology,and the lumen was narrowed,with obvious plaque formation.NDG endometrium is smooth,FDG endometrium is thickened,and subintimal macrophages are infiltrated.Conclusion: The rabbit models of carotid and aortic arch atherosclerosis were successfully established by ultrasonography and pathology.2.Study on the effects of resveratrol on atherosclerosis in rabbits using different imaging techniquesObjective: To observe the changes of carotid atherosclerosis with different imaging techniques and the effect of resveratrol on carotid atherosclerosis in rabbits.Methods: Thirty healthy normal New Zealand white rabbits were randomly assigned to control group,high-fat diet +SPL group,and high-fat diet + resveratrol+SPL group.The operation is the same as before.After 3 months of feeding,ultrasound(GE,USA)was used to detect carotid blood flow velocity,stenosis degree and plaque texture.HRMRI(Siemens,Skyra 3.0T,Germany)detected plaque texture and stenosis degree.PET/CT(u MI510)was used to detect the uptake of plaque nuclides.Results: The lumen of the control group was smooth and no plaque formation was observed.Compared with the high-fat diet +SPL group,the carotid blood flow velocity and stenosis degree of the high-fat diet + resveratrol +SPL group were reduced(P>0.05).The plaque thickness,the proportion of soft plaque and mixed plaque were lower(P<0.05),and the plaque texture was more stable.HRMRI showed that the common carotid artery in the control group had normal morphology and no plaque in the lumen.Compared with the high-fat diet +SPL group,the high-fat diet +resveratrol +SPL group reduced the carotid artery stenosis and the plaque in the blood vessels,leading to a reduced degree of stenosis.PET-CT showed that compared with the high-fat diet +SPL group,the nuclide uptake(SUV)in the plaque(stenosis)site of the high-fat diet + resveratrol +SPL group was reduced(P < 0.05).Conclusions: Ultrasound,HRMRI and PET/CT imaging examination have different advantages in evaluating the carotid atherosclerosis model in rabbits.While confirming the presence of carotid atherosclerotic plaque in rabbits,it suggests that the presence of vulnerable plaque can be found.Resveratrol has anti-atherosclerotic effect,has the effect of reducing plaque metabolism and promoting plaque stability.3.Study on the effect of resveratrol on the regulation of Smads signaling pathway against atherosclerosisObjective: To investigate the effect of resveratrol on Smads signaling pathway of carotid atherosclerosis in rabbits and its anti-arteriosclerosis effect through anti-inflammatory.Methods: The control group,the high-fat diet +SPL group,the high-fat diet +resveratrol +SPL group,the carotid artery was dissected after imaging examination,and HE staining was used for pathological examination.The expressions of Smad2,Smad3 and Smad7 were observed by Western-blot and immunohistochemical technique,and the expressions of RAM11,Actin and CD31 were observed by immunohistochemical technique.Quantitative analysis of western-blot results,IPP software was used to measure the Area and IOD values of the immunohistochemical results.SPSS19.0 software was used for data statistics and t test.Results: HE staining showed that the tube wall of the control group was smooth without plaque formation,and plaque thickness was reduced in the resveratrol +high-fat diet +SPL group compared with the high-fat diet +SPL group(P<0.05).Immunohistochemistry and Western-blot showed that Smad2,Smad3 and Smad7 were increased in the high-fat diet +SPL group compared with the control group.The expression of Smad2 and Smad3 in the high-fat diet + resveratrol +SPL group was lower than that in the high-fat diet +SPL group,and the expression of Smad7 was consistent.The quantitative analysis of CD31,RAM11 and Actin in the high-fat diet + resveratrol +SPL group was significantly lower than that in the high-fat diet +SPL group(P<0.05).In addition,quantitative analysis of CD31,RAM11 and Actin in the high-fat diet + resveratrol +SPL group and the high-fat diet +SPL group were all higher than those in the control group(P<0.05).Conclusions: High-fat diet can activate the Smads signaling pathway,and resveratrol may enhance the expression of Smad7 by inhibiting the expression of Smad2 and Smad3,and inhibit the infiltration of macrophages,smooth muscle hyperplasia and angiogenesis for anti-atherosclerosis effect.4.Effect of resveratrol on blood lipids and Lp-PLA2 in rabbits with atherosclerosisObjective: To investigate the effect of resveratrol on serum lipids and Lp-PLA2 in rabbit model of atherosclerosis.Methods: The rabbits were randomly divided into three groups: NDG,FDG and RFG.Experiment before the heart blood,eat after 3 months,for cardiac blood again,by using chemiluminescence detection of blood lipids,the dry fluorescence immunoassay detection Lp-PLA2 level,blood after anesthesia in aortic arch,line HE staining to observe the pathological changes,observe aorta intima thickness ratio,smooth muscle layer thickness,the lining/smooth muscle layer,data statistical analysis adopt SPSS19.0 software,carries on the t test.Results: There were statistically significant differences in TC,HDL-C,LDL-C and Lp-PLA2 in the last three groups(P<0.05),and the values of TC,LDL-C and Lp-PLA2 in RFG were lower than those in FDG group(P<0.05).Conclusion: Resveratrol can resist atherosclerosis by regulating blood lipid and inhibiting Lp-PLA2.