To Study The Mechanism Of Maixinkang Anti-atherosclerosis Of Apolipoprotein E Gene Knockout Mice And Its Prevention Of The Foam Cell Formation

Objective:To study the etiological factor,pathogenesis and new therapeutics ofatherosclerosis by means of benefiting vital energy to activate blood,remove stasis,anddiscuss the mechanism of atherosclerosis and the theoretical basis of the intervention innew therapeutics of benefiting vital energy,toxin-resolving and blood-activating.Toexplore the pharmacologic action and effect of Capsule Maixinkang on Atherosclerosis andinvestigate the mechanism of maixinkang to prevent macrophages transforming into foamcell.Method:To study the etiological factor and pathogenesis of atherosclerosis,to sumup the result of the observation Capsule Maixinkang on atherosclerosis,hyperlipidemiaand the experiments of effect of Maixinkang to high fat diet rat and ApoE(-/-)mice.Analyze the composition of Maixinkang,explore the pharmacologic action and effect ofMaixinkang on atherosclerosis.Summarize the academic experience in treatingatherosclerosis by benefiting vital energy,activating blood to remove stasis,andtoxin-resolving.Six weeks old ApoE(-/-) mice were divided randomly into four groups,inwhich lavaged with maixinkang,lovastatin,rosiglitazone and saline for two weeks,half ofwhich lavaged for eight weeks,while normal control group which is composed of sixweeks old C57BL/6J mice lavaged no medicine.Half of mice in all groups is put to deathrespectively,collect venous blood and arteriae aorta after eight and twelve lavaged.The areaof Aorta atherosclerosis which observed under light microscope and electron microscopewas analyzed by image analysis system.The expression of PPARγ,CD36,ABCA1 mRNA was detected hybridization in situ.The PPARγ,CD36,ABCA1 protein was evaluatedby immunohistochemistry.The serum concentration of TNF-α,IL-1βwere detected byELISA.2.Peritoneal macrophages derived from ApoE(-/-) mice were incubated withox-LDL and blood serum containing medicine for 72 hours.The peritoneal macrophageswere divided into six groups according to blood serum containing different medicine:Maixinkang,Lovastatin,rosiglitazone,saline and control group.The congcentration ofCa²⁺ intra-cellular was observed and analyzed with Ca²⁺ image analysis system andconfocal laser scanning microscope.Intracellular total cholesterol and free cholesterol(FC) was measured by high performance liquid chromatography (HPLC),ABCA1 andPPAR),protein was detected in immunohistochemistry and semiquantitative analyzed byimage analysis system.CD36 protein was detected by flow cytometry.The expression ofPPAR3,,CD36,ABCA1 mRNA was detected hybridization in situ.Result:1.Maixinkang,relieve and regress atherosclerosis plaque in ApoE(-/-) miceaorta,even for advanced plaques by reducing blood lipid,anti-inflammation,anti-oxidation,protecting vascular endothelium,inhibiting the degradation of extracellular matrix,whichcertificate the therapeutics effect of benefiting vital energy,toxin-resolving andblood-activating has highly effective to prevent atherosclerosis.2.Maixinkang relieveultramicrostructure pathological changes,prevent atherosclerosis and reduce atheroscleroticplaque in aorte.The expression of ABCA1 and PPARγmRNA increasing obviously inMaixinkang group,Lovastatin,rosiglitazone group,thus,.the expression of ABCA1 andPPARγprotein increasing in Maixinkang,Lovastatin,rosiglitazone group,while CD36mRNA and protein expressed less than control group and saline group.The bood serumconcentration of TNF-a,IL-1 in Maixinkang,Lovastatin,rosiglitazone group lowersignificantly than saline and control group.The concentrition of Ca²⁺ of the activatedmacrophages increased fast in control group and saline group more than Maixinkang,Lovastatin,rosiglitazone group.The expression of ABCA1 and PPARγmRNA increasingobviously in Maixinkang,Lovastatin,rosiglitazone group,thus,the expression of ABCA1and PPARγprotein increasing in Maixinkang,Lovastatin,rosiglitazone group,whileCD36 mRNA and protein expressed less than control group and saline group.The bood serum concentration of TNF-α,IL-1βin Maixinkang,Lovastatin,rosiglitazone grouplower significantly than saline and control group.The lever of intra-cellular Cholesteroland cholesteryl ester of Maixinkang,Lovastatin,rosiglitazone is lower than saline andcontrol group in 48h and 72h cultrued.Conclusion:The therapeutic of Combination of benefiting vital energy,toxin-resolving and blood-activating method in anti-atherosclerosis has important clinicaland academic significance.Our research indicated that Maixinkang has the fouction ofinhibit the formation of foam cell,anti-inflammatory,anti-inflammatory,down regulationof macrophages chematropism,anti-atheroscierosis and stabilize atherosclerotic plaque byincreasing the expression of ABCA1 and PPARγmRNA and protein intra-cellular,reducing the level of TNF-αIL-1βin blood serum and culture medium,decreasing theCholesterol and cholesteryl ester and degrading the calcium overload intra-cellular.