| Objective:Coronary heart disease (CAD) is the leading cause of death inthe elderly as a systemic fibrous hyperplasia inflammatory diseases. Modernmedicine consider the basis of coronary heart disease is coronaryatherosclerosis (atherosclerosis,AS). Multiple factors are involved in thepathological process of AS, such as vascular endothelial damage, lipidinfiltration, platelet adhesion and aggregation, inflammatory cell infiltration,expression of the immunity factor, oxygen free radicals damage, excessiveproliferation of vascular smooth muscle, local thrombosis, calcium overload invascular smooth muscle cells (VSMC) caused by the increasing influx ofcalcium and other pathological link related. These mechanisms play the keyroles in the process of atherosclerotic plaque erosion and disintegration. Inrecent years, people have recognized that drug therapy against various targetsin atherosclerosis is expected to become one of the effective means for theprevention and treatment of AS.Shenshao oral liquid is the traditional Chinese medicine for the clinicaltreatment of AS.According to the pre-study of our group, it can develop itsprotective effect on myocardial cell by reducing the level of serum vascularendothelial cell adhesion molecule-1and E-selectin, releasing the oxygen freeradicals oxidative damage, protecting the structure and function of myocardialmitochondria and so forth. However, whether Shenshao oral liquid hasremedial effect on atherosclerosis is unclear. In addition, its protectivemechanism on cardiovascular system is still unknown.This study was divided into three parts. By constructing the model of ratatherosclerosis, it explored the regulatory effect of Shenshao oral liquid onlipid levels and oxidative stress. In the mean while, it observed whetherShenshao oral liquid can inhibit the expression of inflammatory factors. These experiments may provide more theoretical foundation and experimental basisfor the treatment on AS of this drug.Methods:The50rats were randomly divided into5groups:1) control group;2)model group;3) Shenshao oral liquid low-dose group;4) Shenshao oral liquidhigh-dose group;5) simvastatin group. Normal group was given conventionalbreeding. The model group and treatment group were given high fat diet+VD360million IU/kg/d by intraperitoneal injection of3days. After4weeks,Shenshao oral liquid high-dose was given the medicine under the dosage of8ml/d. While the low-dose group was given the medicine under the dosage of4ml/d. The simvastatin group was given the medicine under the dosage of5mg/kg/d. Other groups were given saline. The rats were sacrificed after drugfor8weeks. The blood was drawn from femoral artery of rats. The serum totalcholesterol (TC), triglycerides(TG) and low-density lipoprotein cholesterol(LDL-C) were detected by enzymatic assays with use of an automatedbiochemical analyzer. The malondialdehyde (MDA),glutathione peroxidase(GSH-PX), catalase（CAT）and superoxide dismutase (SOD) were detected byELISA assay to observe the effect of Shenshao oral liquid on oxidative stressindicators. Take a fixed embedding of the complete aorta, HE staining of rataortic atherosclerotic plaque formation.The expressions of Intercellularadhesion molecule-1（ICAM-1）,Vascular cell adhesion molecul（eVCAM-1）,Interleukin-1β（IL-1β）,Interleukin-17A（IL-17A）, Interleukin-23（IL-23）and α-smooth muscle actin（α-SMA）were detected by immunohistochemicalstaining. The expressions of tissue ICAM-1, VCAM-1, IL1β, IL-17A, IL-23protein were detected by western blot.Results:1HE staining to atherosclerosis rat model.HE staining showed that the aortic lumen is stenosis in the aorticatherosclerosis model group.The tunica intima is thickening and protrudinginto the lumen.We can observed that a large number of smooth muscle cellproliferation, plaque and accumulation of foam cells under the tunica intima.We can found calcium deposition and foam cell necrosis area in thebottom of the plaque. The elastic fiber degenerating and tunica mediashrinking leading to the disorder of the three-tier structure of the arterial wall.The tunica intima and tunica media have a large number of plaques andinflammatory cell infiltration. In the Shenshao treatment group, especially thehigh-dose group, atherosclerotic plaque lesions is reduced significantly.2The effect of Shenshao oral liquid on lipid levels of AS rats.Compared with the control group, the As model groups significantlyup-regulate serum TC,TG,LAL-C levels (P<0.05). Compared with the modelgroup, the level of serum lipid in Shenshao oral liquid treatment groupdecreases significantly,and the level of serum lipid in high-dose Shenshao oralliquid group is obviously lower than that of the low-dose Shenshao oral liquidgroup.(P<0.05).There is no significant difference between the high-doseShenshao oral liquid treatment group and the simvastatin group(P>0.05).3The effect of Shenshao oral liquid on the oxidative stress in AS rats.Comparison with the control group, AS model group significantlyincreased serum MDA levels and decreased GSH-PX,CAT and SOD levels(P<0.05). Each treatment groups significantly decreased serum MDA levelsand increased GSH-PX,CAT and SOD levels. There is no significantdifference between the high-dose Shenshao oral liquid treatment group and thesimvastatin group (P>0.05). The level of MDA in high-dose Shenshao oralliquid group was significantly lower than that of the low-doseShenshao liquidgroup（P<0.05）. The level of GSH-PX,CAT and SOD in high-dose Shenshaooral liquid group was significantly higher than that of the low-doseShenshaoliquid group（P<0.05）.4The regulation of Shenshao oral liquid on the expression of inflammatorycytokines in AS rats.We examined expression of ICAM-1, VCAM-1, IL-1β, IL-17A andIL-23by immunohistochemical staining. These proteins showed slightstaining in the control group. The expression of ICAM-1, VCAM-1, IL-1β,IL-17A and IL-23was robustly up-regulated, however, in the atherosclerosis group, as evidenced by strong staining. Compared with the atherosclerosisgroup, the expression of ICAM-1, VCAM-1, IL-1β, IL-17A and IL-23wassignificantly reduced in the low and high dose Shenshao decoction groups, aswell as in the simvastatin-treated group.The results of western blot show that compared with control group, theexpressions of ICAM-1, VCAM-1, IL1β, IL-17A and IL-23increasedsignificantly in atherosclerotic model group.It revealed that the expression ofICAM-1, VCAM-1, IL-1β, IL-17A and IL-23increased3.07-,2.92-,2.42-,3.21-and2.34-fold, respectively, in the atherosclerosis group, as compared tothe control group (all p<0.05). The expression of ICAM-1,VCAM-1, IL-1β,IL-17, and IL-23were reduced by57.65%,52.05%,65.7%,59.5%and50.43%respectively, in the high dose Shenshao liquid group, as compared tothe atherosclerosis group.while in the low dose Shenshao oral liquid groupwere reduced by64.16%,71.92%,84.3%,70.72%and62.39%, as comparedto the atherosclerosis group.Conclusion:1Application of vitamin D3+high-fat diet can establish the AS rat model.Shenshao oral therapy can attenuate the progression of atherosclerotic plaques.2Shenshao oral liquid may develop its anti-atherosclerosis effect byregulating the lipid level, inhibiting oxidative stress and inflammatoryresponses. |
PDF Full Text Request