| Objective: Using high-fat diet with positioning of a soft silicone collar around the rabbits’ right carotid artery established early atherosclerotic models, and intervened with drugs, observed the morphological changes of the carotid artery, the changes of blood lipids, the correlation factors’ expressions of angiogenesis and inflammatory, et al. further explored the effect of a treatment proposal, which consisted of atorvastatin,tongxinluo,and aspirin, on the inflammatory mechanisms of vasa vasorum angiogenesis, this study will provide a new idea for combination traditional Chinese and Western medicinein preventing and treating early atherosclerotic lesions.Methods:1 Effects of Tongxinluo combined with Atorvastatin and Aspirin(ATS) on early atherosclerotic New Zealand rabbitsNinety healthy New Zealand rabbits with half males and half females were fed adaptively for 2 week, and then divided into 6 groups randomly: control group, model group, tongxinluo(TXL) group, atorvastatin(ATO) group, aspirin(ASP) group, golden triangle(ATS) group. n=15.The control group was fed with normal diet, TXL, ATO, ASP, ATS groups’ right carotid arteries with the silicone tube, and were fed with high-fat diet.Whether related papers or the body surface area algorithm of calculating human and animals’ reasonable dose: the suspension of corresponding concentration was consist of tongxinluo superfine powder, atorvastatin and aspirin which dissolved in 0.5% sodium carboxymethyl cellulose(CMC-Na). The TXL group, ATO group and ATS group were given suspension of tongxinluo superfine powder, atorvastatin and aspirin, dosage: 0.3g·Kg-1·d-1, 2.5mg·Kg-1·d-1, 12mg·Kg-1·d-1, the golden triangle group were given suspension of tongxinluo superfine powder(0.3g·Kg-1·d-1), atorvastatin(2.5mg·Kg-1·d-1) and aspirin(12mg·Kg-1·d-1), the control group and the model group were given 0.5% CMC-Na by 3ml·Kg-1, The experiment lasted for 4 weeks.Tissue slice of carotid artery was stained with HE and The pathological morphological changes in carotidartery were observed under light microscope. The change of blood liquid was detected by biochemical assay. Color microsphere method to detect the blood perfusion changes of the cartoid artery microvascular.2 Interventional effects of Tongxinluo combined with Atorvastatin and Aspirin(ATS) on the angiogenesis of vasa vasorum in early atherosclerotic New Zealand rabbitsOn the foundation of creating the early stage of atherosclerosis modelsuccessfully, to further explore effects of tongxinluo combined with atorvastatin and aspirin(ATS) on the the level of factors, which associated with angiogenesis on the adventitia, The methods of modeling, grouping,administration and taking material were the same as part 1. At the end of the test, immunohistochemical staining was used to detect the protein expression of CD34 around the carotid artery adventitia.VEGF,VEGFR-2 gene and protein’ s expression in the cartoid artery were detected by Real-time PCR and Western Blot.3 Interventions of Tongxinluo combined with Atorvastatin and Aspirin(ATS) on the mechanisms of the adventitia inflammatory in early atherosclerotic New Zealand rabbitsOn the foundation of previous two parts experiments, this part further explored the effects of tongxinluo combined with atorvastatin and aspirin on the level of inflammatory factors, the possible inflammatory mechanism and molecular signal transduction pathways, which related with carotid artery wall vasa vasorum angiogenesis.The methods of modeling, grouing, administration and taking material were the same as first part. At the end of the test, Real-time PCR were used to detect the expressionof MCP-1,IL-8, NF-κB,and Western Blot were used to detect the expression of NF-κB.The immunohistochemical staining was used to detect theprotein expression of MCP-1,IL-8,NF-κB around the carotid artery adventitia.The expression of IL-6 in the carotid artery adventitia was detected by ELISA.Results: 1 Effects of Tongxinluo combined with Atorvastatin and Aspirin(ATS) on early atherosclerotic New Zealand rabbits 1.1 The pathomorphism features of carotid artery in various groupsControl group: the shape of the separated endothelial cell are integral, nucleus are flat, and fit closely with the inner elastic plate, aligned smooth muscle.Model group: there is obviously exfoliate of endothelial cells in this group, and Intimal partly thickening in medication administration groups, the injury of carotid artery in TXL, ATO, ATS groups were significantly reduced. 1.2 The levels of blood lipids in various groupsExcept ASP group, compared with the control group, the contents of serum TC, TG, LDL-C in model group increased obviously(P<0.01). The contents of serum TC, TG, LDL-C in TXL group, ATO group and ATS group were all obviously lower than model group(P<0.01, P<0.05). The contents of serum TC, TG, LDL-C in ATS group were lower than TXL group and ATO group( P<0.01), there was no significant difference in the contents of serum TC, TG, LDL-C, HDL-C between TXL group and ATO group(P>0.05). 1.3 Comparison of microvascular blood flow volume of the carotid artery wall in various groupsCompared with the control group, the microvascular blood flow volume of the carotid artery wall in the model group were significantly increased(P<0.01). Those in each medication administration group were lighter than those in the model group(P<0.01). In the ATS group, the microvascular blood flow volume of the carotid artery wall was lower than that in the TXL group, ATO group and ASP group(P<0.01). The microvascular blood flow volume of the carotid artery wall in different subgroups showed no significant difference(P>0.05). 2 Interventional effects of Tongxinluo combined with Atorvastatin and Aspirin(ATS) on the angiogenesis of vasa vasorum in early atherosclerotic New Zealand rabbits 2.1 The positive expression of CD34 of the cartoid artery adventitiaThe CD34 antibody positive stainings of the wall in the model group has a progressive increase compared with the comtral group. Compared with the model group, the content of CD34 in the other medication administration groups were decreased. 2.2 The gene and protein expressions of VEGF, VEGFR-2 of the carotid artery tissue in various groupsCompared with the control group, the gene and protein expressions of VEGF, VEGFR-2 in the model group were significantly increased(P<0.01), but those in each medication administration group were lighter than those in the model group(P<0.01,P<0.05). In the ATS group, the gene and protein expressions of VEGF, VEGFR-2 in cartoid artery were lower than that in the TXL group, ATO group and ASP group(P<0.01,P<0.05). Compared with the ASP group, the protein expression of VEGFR-2 were significantly decreased in the TXL group and ATO group(P<0.05). The gene and protein expressions of VEGF, VEGFR-2 in different subgroups showed no significant difference(P>0.05). 2.3 Tthe content of ES of the cartoid artery adventitia tissue in various groupsCompared with the control group, the content of ES in the model group was significantly decreased(P<0.01). but this in each medication administration group was highter than this in the model group(P<0.01). In the ATS group, the content of ES was higher than that in the TXL group, ATO group and ASP group(P<0.01). Compared with the ASP group, the content of ES was significantly increased in the TXL group and ATO group(P<0.01). The content of ES in the TXL group and ATO group showed no significant difference(P>0.05). 3 Interventions of Tongxinluo combined with Atorvastatin and Aspirin(ATS) on the mechanisms of the adventitia inflammatory in early atherosclerotic New Zealand rabbits 3.1 The comparison of MCP-1m RNA, IL-8m RNA and IL-6 expressions of the carotid artery tissue in various groupsCompared with the control group, the expressions of MCP-1m RNA, IL-8m RNA and IL-6 in the model group group were significantly increased(P<0.01). Compared with model group, the expressions of MCP-1m RNA, IL-8m RNA and IL-6 were decreased in each medication administration group(P<0.01,P<0.05). In the ATS group, the expressions of MCP-1m RNA, IL-8m RNA and IL-6 were lower than that in the TXL group, ATO group and ASP group(P<0.01).No significant differences between the 3 group were observed(P>0.05). 3.2 The gene and protein expressions of NF-κBof the cartoid artery tissue in various groupsCompared with the control group, the gene and protein expressions of nuclear NF-κB in the model were significantly increased(P<0.01), but those in each medication administration group were lighter than those in the model group(P<0.01). In the ATS group, the gene and protein expressions of nuclear NF-κB were lower than that in the TXL group, ATO group and ASP group(P<0.01,P<0.05).The gene and protein expressions of Nuclear NF-κB in different subgroups showed no significant difference(P>0.05). 3.3 The positive expression of MCP-1, IL-8and NF-κB of the cartoid artery adventitia in various groupsThe MCP-1, IL-8and NF-κB antibody positive stainings of the wall in the model group have a progressive increase compared with the model groups. The content of MCP-1, IL-8and NF-κB were decreased compared with the other medication administration groups.Conclusion:In this research, using high-fat diet with positioning of a soft siliconecollar around the rabbits right carotid artery established early atherosclerotic models, and intervened with drugs, we confirmed the Golden Triangle(ATS) project can inhibit the development of AS, decrease the artery wall microvascular blood flow volume, reduce the expressions of proangiogenic factors(CD34,VEGF,VEGFR-2), increase the expression of angiogenesis-inhibiting factor(ES). Indirectly demonstrating the project can reduce carotid artery wall vasa vasorum angiogenesis, the development of AS was effectively delayed in carotid artery, its role was superior to three drugs alone. The mechanism was related to the reduction of inflammatory cytokine expression and Inhibition of vascular adventitia inflammation, and maybe the effection on the NF-kappa B signaling pathway was one of the upstream mechanisms to inhibite vascular adventitia inflammation. |
PDF Full Text Request