| Background:Atherosclerosis is the pathological basis of acute coronary syndrome and acute stroke events. Although the percutaneous coronary intervention and coronary artery bypass grafting can timely open the culprit artery,form bypass circulation and improve blood supply to save the lives of patients, but can not interfere and reverse the pathological process of atherosclerosis. Therefore, the application of anti atherosclerosis drug treatment is still the fundamental prevention and treatment. At present, the western medicine aim at antiplatelet, antioxidation, stabilizing and reversing atherosclerotic plaque, reducing blood lipid and other aspects of the treatment of atherosclerosis. But during the treatment, there are some patients can not tolerate long-term treatment with drug reactions. Therefore, we should study the therapeutic targets of atherosclerosis in order to find more ways to alleviate the condition of patients with atherosclerosis. In recent years, with the development of the research field of the molecular level of cardiovascular disease, a large number of studies have indicated that the atherosclerosis disease is always running through the cell apoptosis and more data suggest that apoptosis is a major factor in the progression of atherosclerosis. Therefore, the treatment of anti-apoptosis can effectively inhibit the progression of atherosclerosis.Studing anti- apoptosis drugs used in clinical practice can better for treatment of different therapeutic mechanism. The theory of Chinese medicine consider congenital deficiency and postnatal malnutrition, especially diet and emotional imbalance cause the pathological changes of the body. The phlegm and blood stasis in the chain of cardiovascular events always exists, which is the reason that the angina pectoris can not be cured. Guided by this theory, the creation of "Fuxie" according to etiological treatment and combined with Chinese medicine treat-ment put forward the "Yiqi Huoxue Huatan" treatment of disease and prepare the Shenhong Tongluo granule. Clinical observation found that Shenhong Tongluo granule can effective cure the phlegm turbidity obstructing the collaterals syndrome and reduce the incidence of cardiovascular events such as myocardial infarction, sudden death, cardiac angina and so on and improve the quality of life. Therefore, with the breakthrough point of cell apoptosis in coronary atherosclerosis,studing the effect of Shenhong Tongluo granule on atherosclerosis plaque and cell apoptosis can provide reliable experimental evidence for the theory of "phlegm and blood stasis". Objective:To observe the pathological changes and apoptosis of the aorta atherosclerotic plaque of apolipoprotein E gene knockout mice by Shenhong Tongluo granule and to investigate the effect of Shenhong Tongluo granule on atherosclerosis. Method:Choose 6 week old healthy male apolipoprotein E gene knockout(Apo E-/-) 30 mice as experimental subjects, weight 25+0.3g. After 1 week routine feeding 30 mice were randomly divided into high fat diet group, high fat diet+ Shenhong Tongluo granule group and normal diet group with 10 rats in each group and offered two groups of mice on a high-fat diet, high fat diet by(1% cholesterol +8% butter +7.5% egg yolk powder +83.5% ordinary feed + bear deoxycholic acid(2mg/kg / d) + propylthiouracil(4mg/kg/d). After 4 weeks of feeding, three groups of mice were given 0.9% saline, Shenhong Tongluo granule(10.12g/kg/d) and 0.9% saline for 8 weeks.After 13 weeks each group were extracted eye blood and use the enzyme linked immunosorbent assay to measure triglyceride, serum total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol. And then kill the mice,got the arch of aorta tissue to make Hematoxylin eosin staining for observing degree of atherosclerosis and histopathological change of aorta. Immunohistochemical staining was used to detect the expression of Caspase-3 protein of apoptosis related genes. Td T-mediated d UTP nick end labeling was used to detect apoptosis in atherosclerotic plaque. Result:Compared with the normal diet group, high-fat diet group, TG, TC, LDL-C index were significantly increased, the difference was statistically significant(P<0.05). Compared with the high fat diet group, the LDL-C and TC of high fat traditional Chinese medicine infusion group were significantly lower and HDLC was significantly higher, and the difference was statistically significant(P<0.05). Light microscope observation of pathological section of Apo E-/- mice aortic arch found the area of atherosclerotic plaque area and the ratio of the lumen area of the vascular lumen in the high fat traditional Chinese medicine perfusion group were significantly higher than that of the common diet group, and the difference was statistically significant(P<0.05). Detection of cell apoptosis in aortic arch atherosclerotic plaque by TUNEL and the number of apoptotic cells of the Chinese medicine infusion group was significantly decreased compared with the high fat diet group and the apoptotic index was also significantly reduced, and the difference was statistically significant(P<0.05). Apoptotic cells are concentrated in the lipid core under the atherosclerotic fibrous cap. The expression of caspase-3 protein in atherosclerotic plaque was detected by immunohistochemistry. The results showed that the expression of caspase-3 protein in high fat diet group was the highest and the staining of atherosclerotic plaque was deepened and shallow in the high fat traditional Chinese medicine perfusion group. Conclusions:1、Shenhong Tongluo granule can reduce serum total cholesterol(TC), triglyceride(TG) and low density lipoprotein cholesterol,which can prevent and cure diseases by reducing risk factors for atherosclerosis.2、Early intervention treatment of Shenhong Tongluo granule can control the progression of the atherosclerotic plaque of apolipoprotein E gene knockout mice in high fat diet, and play a role in the treatment of atherosclerosis.3、Shenhong Tongluo granule can inhibiti cell apoptosis in the atherosclerotic plaque and reduce the expression of Caspase-3 apoptosis related protein to control the atherosclerotic progress. To provide a new theoretical basis for the prevention and treatment of atherosclerosis by the theory of "phlegm and blood stasis". |
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