| Objective:To study the therapeutic effect and mechanism of cooling blood and dissipating blood stasis method on rabbit model of atherosclerosis(AS).Method:1.Establishing a high-fat AS animal model:selecting New Zealand rabbits,clean grade,male(one month old).The cholesterol content in the high-fat diet was set at 0.20%.After balloon dilation,the high-fat diet lasted for 16 weeks.2.Animals were divided into 6 groups,10 in each group,which were normal control group(group A),simple high-fat control group(Bl group),atorvastatin group(B2 group),Qingxin Tongmaiyin(QXTM)low dose Group(B3 group),QXTM middle dose group(B4 group),QXTM high dose group(B5 group).Groups B3,B4 and B5 were administered with Qingxin Tongmaiyin 3.33 g·kg-1 d-1,6.66 g kg-1·d-1,13.32 g kg-l·d-1,respectively;B2 group was administered with atorvastatin calcium suspension at a dose of 2.6 mg kg-1·d-1;Group A and group B1 were intragastrically administered with normal saline once a day for 4 weeks.3.At the end of the 16th week,anesthetized animals,carotid blood was collected,centrifuged that at 1500 rpm for 15 minutes,serum was separated,and total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)were measured by biochemical analysis.High density lipoprotein cholesterol(HDL-C)levels;The levels of inflammatory factor C-reactive protein(CRP),interleukin-1(IL-1),tumor necrosis factor-a(TNF-a)and interferon-β(IFN-β)were detected by enzyme-linked immunosorbent assay.4.The abdominal aorta was isolated from the animal,and the lesions of AS were observed by the naked eye.Pathological sections and HE staining were performed.The pathological observation of AS was performed by Daley s fractional microscopy.5.Quantitative reverse transcription-PCR method was used to detect the expression of apoptosis signal molecules bax,Bel-2,p38 MAPK,ATF-6 and caspase 3 in arterial plaques.Result:1.HE staining results showed that compared with group A,foam cell necrosis and fibrous cap calcification were observed in the endothelium of group Bl,lipid accumulation was observed in the vascular lumen,and the number of smooth muscle cells in the middle layer of the blood vessels was reduced,arranged disorderly,and atrophied to varying degrees.The vascular membrane was thinned,and new capillaries and connective tissue hyperplasia were observed in the outer membrane,and cell necrosis visible in the plaque.In the B2?B3.B4.and B5 groups,different degrees of lipid deposition were observed in the arterial intima,but compared with the model group,the intima of the Chinese medicine group was intact and the fibrosis was reduced.2.The results of biochemical experiments showed that compared with group A,TC,TG,HDL-C and LDL-C in group B1 were significantly increased(p<0.01).Compared with group Bl,TC,TG and LDL-C levels were significantly down-regulated in B2 group,HDL-C levels were significantly increased(p<0.01);TC,TG and LDL-C levels were slightly down-regulated in B3 group,and HDL-C levels were slightly lower.There was no significant difference in the levels of TC,TG,and LDL-C in the B4 group(p<0.01 or p<0.05),and increased in the B3 group.HDL-C level,but the difference was not statistically significant(p>0.05);TB,TG,LDL-C levels were significantly down-regulated in B4 group(p<0.01),HDL-C levels were significantly increased(p<0.01).3.Elisa results showed that compared with group A,the levels of CRP,TNF-α,IL-1 and IFN-P in group B1 were significantly higher(p<0.01),and the levels of serum CRP,TNF-α,IL-1 and IFN-P in B2,B3,B4 and B5 groups were significantly down-regulated(p<0.01).4.PCR results showed that compared with group A,the expression of apoptotic factors bax5 ATF-6,p38 MAPK and caspase-3 mRNA in plaque of group B1 was significantly increased(p<0.01),and anti-apoptotic factor Bcl-2 was obvious.Decrease(p<0.01).Compared with group Bl,the expression of bax,ATF-6,p38 MAPK and caspase-3 mRNA in group B2 was significantly decreased(p<0.01),and the expression of Bcl-2 mRNA was significantly increased(p<0.01).The expression of bax and caspase-3 mRNA in group B3 was decreased(p<0.05),the expression of Bcl-2 mRNA was increased(p<0.05),and the expression of ATF-6 and p38 MAPK was slightly decreased.There was no significant difference compared with group B1(p>0.05);the expression of bax,ATF-6 and caspase-3 mRNA in group B4 decreased(p<0.05 or p<0.01),the expression of Bcl-2 mRNA increased(p<0.05),and the expression of p38 MAPK decreased slightly.There was no significant difference compared with group B1(p>0.05).The expression of bax,ATF-6,p38 MAPK and caspase-3 mRNA in group B5 was significantly decreased(p<0.05 or p<0.01),Bcl-2 mRNA.The expression was significantly elevated(p<0.01).Conclusion:QXTM can improve the progression of atherosclerosis.The mechanism of treatment of atherosclerosis may be related to regulate blood lipid metabolism,reduce apoptotic factors in plaque,inhibit inflammatory response and reduce the expression of inflammatory factors. |
