| Hirudo is a traditional Chinese medicine with complex chemical constituents and widely pharmacologicfunctions. It has been showed that hirudo had the effect of antithrombosis, antitumor, anti-fibrosis and anti-inflummatory and so on. With the development of science and technology, the researchs of hirudo have been doneprofoundly, the research field has been expendied continuously, the pharmacologic functions was much more minningand was widely applied to clinical, hirudo has increasingly become an important drug which can prevent and treatmany diseases, and with prospect of application. As a traditional medicine, hirudo is shinning out new energy throughthe deep and comprehensive studies and it will provide better services to human health.The study of etiology and diagnose method of cardio-cerebrovascular diseases get more and more attention fromChinese and Western medicine scholar, because of the escalating incidence of it. As an important method of modernmedical research, animal model has been used more and more in various diseases, which has played an important rolein the studies of the diagnosis of diseases and mechanism of drug action. Blood stasis syndrome was a majorsymptom in clinic, which include various clinical comprehensive features caused by blood stasis. The modelpreparation of blood stasis ayndrome was almost from the etiology, the methods including single factor and alsocomplex factors. Abundant researches indicated that blood stasis syndrome involving in many physiological andpathological processes, which has a close relation to the hemorheology, abnormal lipid metabolism, and vascularendothelial cell injury and so on.The channel tropism of Traditional Chinese Medicine was the essence of the theory of Chinese Medicine, whichwas accumulated and summarized from a large amount clinic research, which was an important basis for analyzingand describing the effect and mechanism of Chinese medicine, and played an important role in clinic research. So ithas the important significance to study the character, mechanism of medicine, and guide clinic.In this study, the blood stasis syndrome rabbit models induced by complex factors (starvation+high-fat feeding+adrenaline) were established in40d. After modeling, model rabbits randomly divided into four groups, three differentlevels Hirudo groups and model group. Rabbits in Hirudo groups were daily administered with Hirudo (0.30,0.15and0.075g/kg) for30days. The mechanism of Hirodu anti-blood stasis syndrome was studied through determinationof blood lipid lever in serum(Total cholesterol, TG; Triglyceride TC; Low density lipoprotein cholesterol, LDL-C;High density lipoprotein choleaterol, HDL-C), endothelin function indexes in plasma(endothelin-1, ET-1;AngiotensinII, AngII; Nitric oxide, NO), lipid peroxidation indexes in plasma (Malondialdehyde, MDA) and inliver(Malondialdehyde, MDA; Superoxide dismutase, SOD) of blood stasis syndrome rabbits; Realtime PCR wasused to study the levels of expression of Applipoprotein E(ApoE), Low density lipoprotein receptor(LDL-R),Endothelin-1(ET-1) and Endothelial nitricoxide synthase(eNOS) in livers of blood stasis syndrome rabbits. At the same time, the channel tropism of Traditional Chinese Medicine was studed through deteminating the contents ofMDA、SOD in heart, liver, spleen, lung, kidney, stomach, colorectal, intestinal and bile of blood stasis syndromerabbits and nomal rabbits by the obervation method of the active materials in vivo. The main results are as fallows:1) After modeling, the blood lipid of model rabbits was abnomal, compared with control group, the levels of TC,TG, LDL-C in serum of model rabbits were significantly increased(P<0.05), three indexes decreased significantlycompared with model rabbits after treatment of hirudo(P<0.05). The results suggest that hirudo could regulate theblood lipid of blood stasis syndrome rabbits, and in the30d, the low dose of hirudo has the best effect.2) After modeling, the endothelial function of model rabbits was abnomal, compared with control group, thelevels of ET-1, AngII in plasma of model rabbits were significantly increased, the level of NO in plasma wassignificantly decreased (P<0.05), all of three indexes retuned to normal level compared with model rabbits aftertreatment of hirudo (P<0.05). The results suggest that hirudo could regulate the endothelial dysfunction of blood stasissyndrome rabbits, and in the30d, the low dose of hirudo has the best effect.3) After modeling, the lipid peroxidation reaction has happened in vivo of model rabbits, compared with controlgroup, the levels of MDA in plasma and livers of model rabbits was significantly increased, the level of SOD in liverswas significantly decreased (P<0.05), all of two indexes retuned to normal level compared with model rabbits aftertreatment of hirudo (P<0.05). The results suggest that hirudo could inhibit the lipid peroxidation reaction of bloodstasis syndrome rabbits, and in the30d, the low dose of hirud has the best effect.4) After modeling, compared with control group, the gene expression of ApoE and LDL-R in livers of modelrabbits were significantly decreased (P<0.05), the gene expression of ApoE and LDL-R increased significantlycompared with model rabbits after treatment of hirudo (P<0.05). The results suggest that hirudo could regulate thegene expression of gene relevant with blood lipid in livers of blood stasis syndrome rabbits, and in the30d, the highdose and the low dose of hirudo has the best effect to regulate the gene expression of ApoE gene and LDL-Rgene,respectively.5) After modeling, compared with control group, the gene expression of ET-1in livers of model rabbits weresignificantly increased, the gene expression of eNOS in livers of model rabbits were significantly decreased (P<0.05),the gene expression of ET-1decreased significantly, and the gene expression of eNOS increased significantlycompared with model rabbits after treatment of hirudo (P<0.05). The results suggest that hirudo could regulate thegene expression of gene relevant with endothelial function in livers of blood stasis syndrome rabbits, and in the30d,the low dose of hirudo has the best effect.6) Hirudo has definite selectivity to levels of MDA and SOD in each organ whther in blood stasis syndromerabbits or nomal rabbits, which was basically agreement with the traditional channel tropism of Chinese medicine, andthe selectivy of MDA of hirudo in blood stasis syndrome rabbits was much more according with traditional channeltropism of hirudo, the results suggest that hirudo belong to liver meridian, and has definite selectivity to lung, kidney,stomach and colorectal.In summary, the results of this study suggest that complex factors (starvation+high-fat feeding+adrenaline) canestablished model of rabbits blood stasis syndrome, After modeling, compared with control group, the blood lipid,endothelial function of model rabbits was abnomal, and the lipid peroxidation reaction has happened in vivo of model rabbits, the gene expression of ApoE, LDL-R and eNOS gene in livers of model rabbits were significantly decreased,the gene expression of ET-1in livers of model rabbits were significantly increased(P<0.05), After treatment, hirudocould regulate the blood lipid, endothelial function of blood stasis syndrome rabbits, and could inhibit the lipidperoxidation reaction of blood stasis syndrome rabbits, could up-regulate gene expression of ApoE, LDL-R, eNOSgene in livers of blood stasis syndrome rabbits, and down-regulate gene expression of ET-1gene in livers of bloodstasis syndrome rabbits. The results suggest that hirudo has the effect of anti-blood stasis syndrome, the mechanism asfollows:1) Regulate the blood lipid liver and gene expression of blood metabolism related gene LDL-R and ApoE;2)Regulate the endothelial function and gene expression of related gene ET-1and eNOS;3) Inhibit the peroxidationreaction of rabbits. The channel tropism of Traditional Chinese Medicine was studed through deteminating thecontents of active material, the results showed that Hirudo has definite selectivity to levels of MDA and SOD in eachorgan whther in blood stasis syndrome rabbits or nomal rabbits, and the selectivity was basically agreement with thetraditional channel tropism of Chinese medicine. The results suggest that the method of deteminating the contents ofactive material has the feasibility of studying the channel tropism of Traditional Chinese Medicine. |
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