Research On Effects Of Bushenhuoxuehuatan Decoction To Atherosclerotic Plaque Vunerability

AS(artherosclerosis,AS) is common aliment harming health and is the important factor leading to most of CCVD(cardiovascular and cerebrovascular diseases,CCVD),the incidence of AS is increasing recent years.The plaque fracture is the main factor to lead to the clinical end point events of CCVD. There are close ties between the metabolic disorder of lipid,the proliferation and apoptosis of VSMC(vascular smooth muscle cells,VSMC),the promoting apoptosis gene of caspase-3 and the inhibiting apoptosis gene of bcl-2, MMP-2(matrix metalloproteinase-2,MMP-2) and TIMP-2(tissue inhibitor of metalloproteinase-2,TIMP-2)with the plaque vulnerability.There are such theories of TCM(Traditional Chinese Medicine,TCM) that they are the main mechanisms that heat poison,moist heat,blood stasis,phlegm muddy and deficiency of body self or interaction of them lead to AS.The great achievements have been made in the field of researching AS by clearing heat and toxic,removing heat and dampness,promoting blood circulation by removing blood stasis,resolving phlegm and making the Qi,blood and Zangfu strong by tonic methods,but there are fewer research on plaque vulnerability of AS. There are small number specialists think that there is very close relationship between the heat poison and the inflammation mechanism of AS plaque vulnerability and make some studies and get some achievements.It is experience summary comes from the professor Luo Luyi of Guangzhou University of TCM who has been gone in to CVD for many years by the method of notifying the kidney, removing blood stasis and resolving phlegm.There are very good effects by this method on AS and some progress having been made,but the plaque vulnerability has not been involved in.Therefore,this project set out from this point to research the effects of bushenhuoxuehuatan decoction on AS plaque vulnerability.This project set out from the penetration point of plaque vulnerability to research the mechanism of hindering and deferring AS by the bushenhuoxuehuatan decoction,including the clinical study and the animal experiment.1 Clinical Study1.1 Study of Effects of Bushenhuoxuehuatan Decoction on the Patients of CAS Objective:To research the mechanisms that bushenhuoxuehuatan decoction alleviating the symptoms of CAS(carotid arterioles artherosclerosis,CAS) patients and affecting AS progress and AS plaque vulnerability through observing the effects of bushenhuoxuehuatan decoction on the symptoms integral, total efficacy,serum lipid,the carotid IMT(intima-media thicknesses,IMT) and the type and quantity of plaques in carotid arterioles.Methods:96 CAS patients were divided into two groups at random,BSTG (treatment group of bushenhuoxuehuatan decoction) and ACTG(treatment group of atorvastatin calcium,ACTG),48 patients in either group,and to intervene by bushenhuoxuehuatan decoction and atorvastatin separately.Evaluate the alleviating symptom condition and the total efficacy,and detect the serum lipid by ACCA(Automatic Clinical Chemistry Analyser,ACCA),IMT and the type and quantity of AS plaques in carotid of patients by Doppler ultrasound and after 27 weeks.The numerical value were expressed by mean±standard deviation((?)±S) and statistical analyzed with the soft ware of SPSS 11.5.The independent sample t-test was used in comparison among groups,paired sample t-test was used in comparison before and after treatment,Ridit analysis was used in ranked data and chi square test was used in rate comparison.Results:Compared with before treatment,the symptom integral of two groups decreased obviously,there is not marked difference between two groups ((P＜0.02 or P＜0.05)),the total efficacys were very significant and the BSTG was superior to the ACTG.The level of the serum TC and LDL-C of two groups decreased significantly and the level of HDL-C increased obviously(P＜0.01 or P＜0.05).There is no affect on TG in BSTG(P＞0.05),but the level of TG in ACTG in decreased obviously.There is no deference in two groups in HDL-C and TC(P＞0.05).The level of the serum LDL of BSTG increased compared with ACTG(P＜0.05).The IMT of BSTG didn't become thinner but the AS plaques become less obviously(P＜0.01),and there' s no deference between two groups (P＞0.05). Conclusions:The symptoms of CAS patients can be alleviated by bushenhuoxuehuatan decoction,and the total efficacy was very obvious.The serum TC and LDL in CAS patients can be decreased prominently,and the HDL can be increased significantly,and the IMT can not become thinner observably and the AS plaques can become less by bushenhuoxuehuatan decoction.Therefore, there are these effectives of bushenhuoxuehuatan decoction that it can obstruct or retard the progress of the CAS and stable the AS plaques.It is a kind of mechanism to regulate the lipid.1.2 Study of Effects of Bushenhuoxuehuatan Decoction on the Patients of CADObjectives:To research the mechanisms that bushenhuoxuehuatan decoction affects AS plaque vulnerability through observing the effects of bushenhuoxuehuatan decoction on serum lipid,MMP-2 and TIMP-2 of CAD(coronary atherosclerotic heart disease,CAD) patients.Methods:75 CAD patients were divided into two groups at random,there are 25 patients in BSTG and there are 50 patients in ACTG,and to intervene by bushenhuoxuehuatan decoction and atorvastatin separately.Detect the serum lipid by ACCA,MMP-2 and TIMP-2 of patients by ELISA after 3 months.The numerical value were expressed by mean±standard deviation((?)±S) and statistical analyzed with SPSS 11.5 soft ware.The independent sample t-test was used in comparison among groups,paired sample t-test was used in comparison before and after treatment.Results:The level of the serum TC and the LDL-C of two groups decreased significantly in two groups(P<0.01).The level of HDL of BSTG increased obviously(P<0.01).There were not difference in the level of TG and HDL-C of ACTG compare with before treatment.There is no obvious deference of TC between BSTG and ACTG(P>0.05),the level of HDL of BSTG increased and TG decreased obviously compared with ACTG,and the level of LDL-C of ACTG decreased obviously compared with BSTG.The level of the serum MMP-2 and TIMP-2 of two groups decreased prominently compared with before treatment(P<0.01), and there is no deference between the two groups.Conclusions:The serum TC,TG and LDL-C in CAD patients can be decreased prominently,HDL can be increased significantly,MMP-2 and TIMP-2 can be balanced by bushenhuoxuehuatan decoction.Therefore,it can be deduced that there are such effectives of bushenhuoxuehuatan decoction that it can obstruct or retard progress of the CAD and stable the AS plaques.It may be a kind of mechanism to regulate the serum lipid and balance MMP-2 and TIMP-2 by bushenhuoxuehuatan decoction.2 Animal ExperimentsThe effects of Bushenhuoxuehuatan Decoction on the Plaque Vulnerability of AS Model in RabbitsObjective:To research the mechanisms that bushenhuoxuehuatan decoction affects AS progress and plaque vulnerability and investigate the mechanisms causing AS plaque vulnerability through observing the effects of bushenhuoxuehuatan decoction on serum lipid and the expression in AS plaques ofα-SMA(α-smooth musclc actin,α-SMA),PCNA(Proliferating cell nuclcar antigen, PCNA),VSMC apoptosis(VSMC apoptosis,VSMCAP),bcl-2,caspase-3,MMP-2 and TIMP-2 of AS model in rabbits. Methods:60 male rabbits of New Zealand were divided into two groups at random, CG(control group,CG),MG(model group,MG),BSIG(intervention group of bushenhuoxuehuatan deconction,BSIG),ACIG(intervention group of atorvastatin calcium,ACIG),BSTG(treatment group of bushenhuoxuehuatan deconction,BSTG),ACTG(treatment group of atorvastatin calcium,ACTG),and there are 10 rabbits in every groups.The later five groups were made into model,to make AS model by high lipid diet and injecting the vein of ear fringe with BSA(bovine serum albumin,BSA).The deferent interventions were given. CG feed with common fodder.There were no any intervene in MG except made model. BSIG and ACIG were tube feeding through mouth for about 12 weeks until they become successful model.BSTG and ASTG were tube feeding through mouth for 4 weeks after becoming successful model.Analyze the serum lipid level of six groups by ACCA and the expression ofα-SMA,PCNA,bcl-2,caspase-3,MMP-2 and TIMP-2 in AS plaques by HIS(immunol histochemistry staining,HIS).VSMCAP was detected by TUNEL.The numerical value were expressed by mean±standard deviation(X±S) and statistical analyzed with SPSS 11.5.The one way ANOVA was used in comparison among groups,to analyze the relationship among these index by correlated analysis and linear regression.Results:(1) The level of the all serum lipid of the five model groups increased significantly compared with CG(P＜0.01).Compared with MG:the level of the serum TC,TG and LDL-C of BSIG and ACIG decreased and the level of the HDL-C increased significantly(P＜0.01),the level of the serum TC and LDL-C of BSTG and ACTG decreased and the level of the HDL-C decreased significantly(P＜0.01), there is no obvious change of TG in BSTG and ASTG.The level of LDL-C of BSIG decreased significantly compared with ACIG(P＜0.05),and the level of TC of ACIG decreased compared with BSIG(P＜0.05),but there is no obviously deferent in HDL-C and TG between the two groups(P＞0.05).The level of TC in ACTG decreased significantly compared with BSTG and the level of TG decreased and HDL increased obviously in BSTG compared with ACTG(P＜0.05,P＜0.01)),but there is no obviously deferent in LDL-C between the two groups(P＞0.05).(2) The results ofα-SMA,PCNA and VSMC apoptosis expression in AS plaqueThe expression ofα-SMA,PCNA and VSMC apoptosis of AS plaques in five model groups are increased significantly compared with CG(P＜0.001).The expression ofα-SMA,PCNA,VSMC apoptosis are increased significantly in BSIG, ACIG and ACTG compared with MG(P＜0.01,P＜0.05).The expression ofα-SMA and VSMC apoptosis are increased significantly in BSTG(P＜0.001),but there is no change of the expression of PCNA in BSTG compared with MG(P＞0.05).There are no deference that the expression of theα-SMA,PCNA,VSMC apoptosis, between the BSIG and ACIG(P＞0.05).The expression ofα-SMA and VSMC apoptosis are increased in BSTG compared with ACTG(P＜0.05),but there is no difference between two groups.(3) The results of caspase-3and bcl-2 expression in AS plaqueThe expression of caspase-3 increased and bcl-2 decreased significantly in five model groups compared with CG(P＜0.001).The expression of caspase-3 decreased and bcl-2 increased significantly in BSIG,ACIG,BSTG and ACTG compared with MG(P＜0.001).There is no change of the expression of caspase-3 and bcl-2 between the BSIG and ACIG,between the BSTG and ASTG(P＞0.05).(4) The results of MMP-2 and TIMP-2 expression in AS plaqueThe expression of MMP-2 and TIMP-2 increased significantly in five model groups compared with CG(P＜0.001).The expression of MMP-2 and TIMP-2 decreased significantly in BSIG,ACIG,BSTG and ACTG compared with MG(P＜0.001,P＜0.01). There is no difference of the expression of MMP-2 and TIMP-2 between BSIG and ACIG(P＞0.05),between BSTG and ACTG(P＞0.05).Conclusions:(1) The AS model can be made successfully using New Zealand white rabbits through high lipid diet and injecting the vein of ear fringe with BSA.It also illustrates that high lipid diet is very important risk factor leading to AS. The AS progress can be impeded and retarded and the plaque vulnerability can be decreased by regulating serum lipid.(2) The serum lipid of AS model groups can be regulated by bushenhuoxuehuatan decoction.The early intervene to HDL and latter treatment to HDL and TG of bushenhuoxuehuatan decoction is superior to atorvastatin,but the early intervene latter treatment to TC of bushenhuoxuehuatan decoction is inferior to Atorvastatin.(3) There are correlation among theα-SMA,PCNA,VSMC apoptosis,caspase-3, bcl-2,MMP-2 and TIMP-2 and they can be affected each other.(4) The expression ofα-SMA,PCNA,VSMC apoptosis,caspase-3,bcl-2,MMP-2 and TIMP-2 can be regulated effectively by early intervene and latter treatment with bushenhuoxuehuatan decoction.(5) The bushenhuoxuehuatan decoction can effect on AS from multiple target points.It can retard and impede the process of AS and decrease the AS plaque vulnerability by regulating serum lipid,controlling excessive proliferation and apoptosis of VSMC and preventing ECM degradation.The early intervention is superior to latter treatment.(6) There are some correlations between the pathogenesis of kidney deficiency, blood stasis and phlegm stagnation with the process of AS and AS plaque vulnerability.This may be a kind of the pathogenesis of AS.