Effect Of Huatu Tongmai Recipe On Cholesterol Reverse Transporter In Atherosclerotic Rabbits

Atherosclerosis(AS) is an important factor which leads to cardiovascular and cerebrovascular diseases ranking first among all causes of death in developed countries. The growth of people’s lifetime and the aging of population lead to AS with high morbidity and mortality rate. Therefore, it becomes a hot area of research about cause and mechanism, prevention and treatment measures of AS that medical investigator need face and solve. AS is a complex multifactorial pathological process, therefore a single mechanism of action of the drug probably cannot fully inhibit the development of AS. The Chinese compound is based on traditional Chinese medicine theory, focusing on the macro level, comprehensive treatment, overall adjustment in theory, from multi-angle, multi-level and multi-target preventing and treating AS effectively. The presentation of "Pulse turbidity" theory innovate the comprehension of AS pathogenesis and connect with traditional Chinese medicine and modern medicine. Resolving turbidity and dredging collaterals method is established for this syndrome, which has received a good clinical efficacy.Objective:To observe the effects of Huazhuotongmai compound on plaques on arterial wall and lipid in serum and aorta vascular smooth muscle cell of rabbit models of atherosclerosis and investigate the mechanisms of Huazhuotongmai compound for against AS on reverse cholesterol transport(RCT).Experimental methods:There were 40 healthy purebred male New Zealand rabbits divided into four groups randomly:blank control group, model group, simvastatin group, and Huazhuotongmai group. The rabbits of AS were established by simple high-fat feeding method. The blood samples were collected from ear marginal veins before and after drug administration for 14 weeks after abrosia. The thoracic aortas and livers of the animals were taken after execution under anesthesia at the end of 15 weeks. The rabbit fatty streaks and fatty plaques of gross specimen were inspected by visual study. The formation of rabbit arterial plaque was observed by HE staining. The lipidosis in arterial wall was viewed by oil red o staining. The levels of total cholesterol(TC), triglyceride(TG), low-density lipoprotcin(LDL), high-density lipoprotein(HDL) in serum were measured by enzymic method. The levels of TC and free cholesterol(FC) in vascular smooth muscle cell and CD36 in livers were measured by enzyme-linked immuno sorbent assay(ELISA). The expressions of caveolin-1(cav-1), cyclophilin A(CyPA), ATP-binding cassette transporters A1(ABCA1) in arterial wall and scavenger receptor claBs B type I (SR-B I) in liver were measured by western blot(WB). The expressions of mRNA of cav-1, CyPA and ABCA1 in arterial wall, mRNA of SR-B I and CD36 in liver were evaluated by real-time quantitative polymerase chain reaction(real-time PCR).Results:The surface of arterial wall in blank control group is smooth, the layers of structure are clear and the endothelial cells and the smooth muscle cells are continuous without lipidosis. The arterial wall in model group is obviously thickened and covered with lipid plaques obviously. The arterial wall in Huazhuotongmai group is slightly thickened with a little lipidosis. There are no differences between Huazhuotongmai group and simvastatin group. At the end of 14 weeks, TC, TG and LDL in serum in model group are significantly higher than those in blank control group(P<0.01). TC and FC in smooth muscle cells in model group are higher than that in blank control group(P<0.05,P<0.01). TC, TG and LDL in serum in Huazhuotongmai group are lower than those in model group(P<0.05,P<0.01). TC and FC in smooth muscle cells in Huazhuotongmai group are significantly lower than those in model group(P<0.01). After 15 weeks the test results of aorta cells show that compared with blank control group, the expression of cav-1 and cav-1 mRNA in model group are significantly declining(P<0.01). Compared with model group, the expression of cav-1 mRNA in Huazhuotongmai groups is significantly increasing(P<0.01), the expression of cav-1 presents a trend of increasing, which shows no statistical significance(P>0.05). Compared with blank control group, the expression of CyPA in model group is increasiong(P<0.05), the expression of CyPA mRNA presents a trend of declining, which shows no statistical significance (P>0.05). Compared with model group, the expression of CyPA is significantly declining(P<0.01), the expression of CyPA mRNA is significantly increasing(P<0.01) in Huazhuotongmai group.Compared with blank control group, the expression of ABCA1 mRNA in model group is significantly increasing(P<0.01), the expression of ABCA1 presents a trend of declining, which shows no statistical significance (P>0.05). Compared with model group, the expression of ABCA1 mRNA in Huazhuotongmai group is significantly declining(P<0.01), the expression of ABCA1 presents a trend of increasing, which shows no statistical significance (P>0.05).The test results of liver cells show that the expression of SR-B I and SR-B I mRNA in model group declines slightly than those in blank control group, which shows no statistical significance(P>0.05). And compared with model group, the expression of SR-B I mRNA in Huazhuotongmai group presents an upward trend, which shows no statistical significance(P>0.05). Compared with blank control group, the expression of CD36 mRNA in model group is increasing(P<0.05), the expression of CD36 presents an upward trend, which shows no statistical significance(P>0.05). Compared with model group, the expression of CD36 in Huazhuotongmai group presents a trend of declining, which shows no statistical significance (P>0.05).Conclusions:1. Huazhuotongmai compound can reduce lipidosis in artery wall cells and inhibit the plaque formation in artery of rabbits with AS.2. Huazhuotongmai compound can reduce the contents of TC, TG and LDL in serum, TC and FC in artery cells.3. Huazhuotongmai compound can promote the expression of cav-1, cav-1 mRNA, CyPA mRNA and ABCA1, increase transmembrane transport of cholesterol to promote cellular cholesterol efflux, ensure the normal expression of cav-1 and CyPA and reduce the over expression of CyPA, which has an effect on anti-AS.4. Huazhuotongmai compound can inhibit the expression of CD36, promote the expression of SR-B I mRNA and cholesterol metabolism in liver, which has an effect on anti-AS.