| BackgroundInflammation is the main pathology in the Coronary sclerosis.Inflammation biomarkers take part in many patho-physiological course such as inflammation,their levels in the blood can evaluate and predict the inflammate action in coronary sclerosis plaque. TNF-a, E-selectin and ICAM-1are very important to the adhesion between inflammatory cells and endothelium cells.Objecitve1.To observe the clinical epidemiology of CHD,research CHD’s differentiation of symptoms features, test the levels of TNF-a, E-selectin and ICAM-1in different symptoms of CHD,discuss the correlation between them,from the angle of Chinese medicine,provide reference to evaluate condition of CHD by inflammation biomarkers.2.Stimulate HCMEC by TNF-a,produce an inflammatory condition to observe the effects of APS on adhesion between neutrophils and HCMEC,from the angles of protein and gene expression,discuss the effects of APS on endothelium cells inflammatory.Methods1.Investigate243CHD Chinese medicine differentiation syndromes distribution.Observe the change pattern of blood fat and blood coagulation.Test the level of TNF-a,E-selectin and ICAM-1in patients’serum of different syndromes,using medicial statistics methods to investigate those biomarkers diviosion between different Chinese medicine syndromes.Discuss the correlation between TNF-a,E-selectin and ICAM-1and different syndromes,blood fat and blood coagulation.2.Effects of APS on adhesion between neutrophils and endothelium cells and its mechanism:Separate neutrophils from human blood,using TNF-a stimulate adhesion between neutrophils and endothelium cells.using APS to affect this adhesion action.test amount of adhesion cells.discuss the effects of APS on adhesion action.Using Western-blot method to test protein expression of adhesion molecules E-selectin、ICAM-1.Using RT-PCR method to test mRNA of E-selectin,P38and ATF-2.Discuss the effects of APS on adhesion molecules and cells signal transduction,as a result to find the mechanism of APS intervene molecules adhesion.Results1. The distribution of CHD patients with differentiation syndromes orderly are:blood stagnation syndrome> qi deficiency syndrome> phlegm stasis syndrome> qi stasis syndrome> yin deficiency syndrome> yang deficiency syndrome> cold coagulation syndrome.The features of syndromes combination is that three syndromes combination are in the majority, among this, the largest amount is Qi deficiency and blood stagnation syndrome.The second is two syndromes combination. Qi deficiency and blood stagnation syndrome is the basic syndrome in CHD(58.03%of the total patients).2.Correlation between Blood cholesterol and Chinese medicine differentiation syndromes:Blood cholesterol in qi dificency syndromes compare with other syndromes (non-qi dificency syndromes) statistical significance isn’t obvious (P>0.05). TC、LDL-C level in blood stagnation syndrome group is higher than non-blood stagnation syndrome group,while HDL-C level is lower than non-blood stagnation syndrome (P<0.05). HDL-C level in qi deficiency and blood stagnation syndrome is lower than other syndromes (non-qi deficiency and blood stagnation syndrome)(P<0.05). Blood stagnation syndrome is correlated with LDL-C, qi deficiency and blood stagnation syndrome is correlated with HDL-C,phlegm syndrome is correlated with TG,TC and HDL-C.3. Correlation between level of TNF-a、sICAM-1、sE-selectin in serum with CHD differentiation syndromes:TNF-a level of CHD each group is higher than normal group,UAP and AMI group is higher than SAP group(P<0.05). sICAM-1、sE-selectin level of CHD each group is higher than normal group,UAP and AMI group is higher than SAP group (P<0.01). TNF-a,sICAM-1and sE-selectin level of qi deficiency syndrome group,blood stagnation syndrome group, qi deficiency and blood stagnation syndrome group are higher than normal group (P<0.01). TNF-a level in qi dificency syndromes compare with non-qi dificency syndromes statistical significance is obvious (P<0.05).sICAM-1、sE-selectin level in blood stagnation syndromes compare with non blood stagnation syndrome statistical significance is obvious (P<0.05). Correlation between TNF-a and blood stagnation syndrome is extremely obvious (P<0.01) correlation between blood stagnation syndrome and sICAM-1,sE-selectin is obvious(P<0.05). Correlation between qi dificency syndromes and TNF-a,sICAM-1and sE-selectin is obvious (P<0.05).while correlation between qi dificency and blood stagnation syndromes and TNF-a,sICAM-1and sE-selectin is extremely obvious (P<0.01)4.Correlation between adhesion molecules and blood cholesterols and blood coagulation: correlation between sICAM-land FIB.LDL-C.HDL-C is obvious (P<0.05).Correlation between sE-selectin and FIB,LDL-C is obvious (P<0.05)5.Correlation between TNF-a and sICAM-1, sE-selectin:Correlation between TNF-α and sICAM-1,sE-selectin is obvious (P<0.05)6. Effects of APS on TNF-a stimulate adhesion action between neutrophils and HCMEC:HCMEC in modle group was shrinking,fall off,appeared to inflammatory injure.The inflammatory injure in APS group are much lighter than modle group.The adhesion cells amount between neutrophils and HCMEC in modle group is higher than normal group (P<0.01),while in APS group and inhibitor group are much lower than in modle group(P<0.01).7. Effects of APS on TNF-a stimulate adhesion molecules protein expression in HCMEC: ICAM-1protein expression in modle group is much higher than in normal group (P< 0.01).Compare to modle group, ICAM-1protein expression in APS high dose group and inhibitor group is much lower (P<0.01)8. Effects of APS on TNF-a stimulate adhesion molecules mRNA and P38signal pathway in HCMEC:P38and E-selectin mRNA in modle group are much higher than in normal group (P<0.01), ATF-2mRNA in modle group are higher than in normal group (P <0.05).Compare to modle group, P38,ATF-2and E-selectin mRNA in APS high dose group,low dose group and P38inhibitor group are lower (P<0.05)ConclusionThis research found that Qi deficiency,blood stagnation and qi deficiency-blood stagnation syndrome is the most common syndrome. Qi deficiency and blood stagnation syndrome is the basic Chinese medicine syndrome in CHD.Qi deficiency,blood stagnation and qi deficiency-blood stagnation syndrome is obviously correlated with blood cholesterol. Blood cholesterol level are obviously different in different syndrome. Qi deficiency,blood stagnation and qi deficiency-blood stagnation syndrome is obviously correlated with inflammatory biomarkers,which are in according with CHD’s condition.Inflammation take part in the development of CHD, inflammatory biomarkers can predict and evaluate the inflammatory condition of CHD,be reference to disease evaluation. APS can inhibit adhesion between inflammation cells and HCMEC,the mechanism are through the effects of inhibit activation of P38signal pathway,alleviate ATF-2mRNA,to reduce protein and mRNA expression of E-selectin, ICAM-1. |
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