Study On The Effects Of Huanglian Jiedu Decoction To The Rabbit Atherosclerosis Caused By Toxic Heat And The Activity Of AMPK

BackgroundAtherosclerosis is a major risk factor in cardiovascular disease(CVD).With the increasing population in obesity and diabetes,CVD diseases caused by AS such as coronary heart disease(CHD)have become the leading cause of death in the world[1-4].Therefore,hunting for high-efficiency,low-toxic anti-AS agents is one of hot topics in new drugs research.In recent years due to the development of Western medicine to the plateau and its obvious side effects,including the complex mechanism of multi-factor,multi-site,multi-link,multi-targets,multi-network in AS,more and more asian and western scholars turned to the Traditional Chinese Medicine(TCM).However,most scholars only focus on bioactive substances in Chinese herbal medicine such as the study of berberine,ignoring the qualities of herbal medicines and herbal formula.Due to the constraint of research conditions,most scholars choose cell lines in vitro model.Despite efficiency and economy,it could not really reflect inner environment;And the experimental results are still needed to be confirmed.At present,Qing Re Jiedu square-Huanglian Jiedu Decoction(HJD)has been generally recognized as a clear hot alexipharmic in the treatment of AS.But the exact mechanism is still not very clear,and further research is needed.There have been many researches confirmed that the berberine(active substances of coptis)had active effects on the activated protein kinase(AMPK).AMPK has proved to be the target of anti-AS,so activation of AMPK has become a potential target for drug treatment in AS.Berberine can increase the activity of AMPK in liver cells and endothelial cells[5-7].Therefore we conclude that one of the mechanism of HJD anti-AS may be activation of AMPK signaling pathway in liver and vascular endothelial cells,and thus regulating energy metabolism in vivo.ObjectiveTo investigate the effects of HJD on the rabbit atherosclerosis and the activity ofAMPK by establishing atherosclerosis rabbit models by toxic heat and high fat,and reveal the mechanism of HJD on the molecular biological levels,and provide a new experimental data for Clearing Heat and Removing Toxin method on anti-AS.MethodsUnder the same experimental conditions,thirty-two healthy male New Zealand white rabbits were randomly divided into four groups(8 rabbits in each group): normal control group,model group,huanglian jiedu decoction(HLJD)group,metformin(Met)group.The rabbits of all the treated groups were injected LPS by ear margin vein and administered with the same high fat diet for setting up rabbit AS model.The rabbits of HLJD and Met groups were interfered by HLJD(1.6 g/kg/d)and metformin(150 mg/kg/d)at the beginning of the fifth week.The blood lipid levels of each group were measured respectively before and after drug intervention.Stop drugs and fast for 12 hours at the end of the 8th week,then kill the rabbit after collecting blood in ear margin vein,observe thepathomorphology of abdominal aorta and hepatic tissue by HE staining,measure the expression levels of AMPK,P-AMPK in hepatic tissue and AMPK,P-AMPK and P-ACC in artery tissue by Western-blot.Study the formation state of AS and its effects on AMPK activity after drugs intervention.Results1.The blood lipid level of model group was higher than all drug groups and normal control groups,the difference was statistically significant(P <0.01;P<0.05).There was no significant difference among HJD group and Met group in blood lipid change(P>0.05).2.The obvious statistical differences were showed in endomembrane thickness、tunica media thickness and intima-media thickness ratio of the model groups respectively compared with drug groups and normal control groups(P<0.0 1).There was no significant difference among HJD group and Met group about indexes above(P>0.05).3.The aorta and hepatic tissue by H-E staining was under the microscope and didn’t show atherosclerotic plaques in the HJD group and Met group,while atherosclerotic plaques were observed in model group.4.Compared with the model groups,the expression of hepatic tissue AMPK P-AMPK and P-ACC levels of drug groups and normal control groups were higher(respectivelyP<0.01)and the expression of artery tissue AMPK and P-AMPK levels were higher(both P<0.01).There was no significant difference among HJD group and Met group blood about indexes above(P>0.05).ConclusionsHJD can adapt AMPK-ACC intracellular pathways and promote fatty acid oxidation in the liver.At the same time HJD can also increase AMPK activity and protect the function of endothelium.This may be one of the mechanisms of HJD anti-AS.So that the subject reveals the mechanism of HJD on the molecular biological levels.