| Objective:To study the correlation between Platelet membrane glycoprotein PAC-1,CD62p, CD63and acute ischemic stroke (IS).To investigate the platelet membrane glycoprotein PAC-1,CD62p,CD63monitoring platelet activation status and the possibility to predict acute IS condition by observe the changes of the factors mentioned above expression in Patients with acute ischemic stroke (IS).To investigate its correlation with the platelet membrane glycoprotein expression and aspirin resistance (AR) by detecting the platelet aggregation and the Platelet membrane GP Ⅱb HPA-3gene polymorphism of patients with acute IS,in order to supplying the evidence of antiplatelet therapy.Methods:1. Acute IS patient group (n=226),normal control group (n=40) and high-risk control group (n=60).High-risk control group for the combined risk factors for stroke,such as hypertension,diabetes,coronary heart disease risk factors.All the subjects were detected in blood routine test, hepatic function,blood lipids,glucose and other biochemical markers.The patient group is divided into light,medium and heavy three subgroups by NIHSS.Meanwhile, the patients also divided into progressive IS and stable IS with there clinical condition within48hours.2. The platelet membrane glycoprotein PAC-1,CD62p,CD63expression were detected by flow cytometry (FCM),using Adenosine diphosphate (ADP) and arachidonic acid (AA) as an inducer to detect the expression of platelet aggregation rate of all the samples. And detect the platelet membrane GP Ⅱb HPA-3gene polymorphism of all the samples by polymerase chain reaction-restriction digestion fragment length polymorphism analysis method.3. Part of the patient group taking aspirin enteric-coated tablets100mg/d for7days,detect the platelet membrane glycoprotein PAC-1the CD62p,CD63and platelet aggregation expression rate for the second time.4. Observed the changes of expression of platelet membrane protein PAC-1,CD62p,CD63and platelet aggregation rate in different NIHSS cases group.5. Observed the changes of expression of platelet membrane protein PAC-1,CD62p,CD63and platelet aggregation rate in progressive IS and stable IS patients.6. Investigate the correlation between Platelet membrane GP Ⅱb HPA-3gene polymorphism and the expression of platelet membrane glycoprotein PAC-1, CD62p, CD63.7. According to the expression level of platelet aggregation after aspirin treatment,screening patients with AR (According to the experimental results,the ADP as inducers measured platelet aggregation rate>51.35%;AA as an inducer of platelet aggregation measured rate>47.63%;to meet the two is defined as AR).And investigate the correlation between AR and Platelet membrane GP Ⅱb HPA-3gene polymorphism.Results:1.The expression of platelet membrane glycoprotein PAC-1、CD62p、CD63were compared in patient group,high-risk control group and the normal control group.The expression of platelet membrane glycoprotein PAC-1was higher in patient group than the normal control group (P<0.05).The expression of platelet membrane glycoprotein CD62p was higher in patient group than the normal control group (P<0.05).The expression of platelet membrane glycoprotein CD62p was higher in high-risk control group than the normal control group (P<0.05).The expression of platelet membrane glycoprotein CD63was higher in patient group than the high-risk control group and the normal control group (P<0.05),the difference was statistically significant.2. The expression of platelet aggregation (induced by ADP) in patient group and the high-risk group were higher than the normal control group,the difference was statistically significant (P<0.05).The expression of platelet aggregation (induced by AA) in patient group were higher than the high-risk group and the normal control group, meanwhile the high-risk control group were higher than the normal control group, the differences were statistically significant (P<0.05).3. The expression of platelet membrane glycoprotein PAC-1、CD62p、CD63and platelet aggregation induced with ADP and AA were no significant difference in different NIHSS subtypes (P>0.05).4. The expression of platelet membrane glycoprotein PAC-1、CD62p、CD63and platelet aggregation in patient group were significantly decreased afeer aspirin therapy,and the difference was statistically significant (P<0.05).5. The expression of platelet membrane glycoprotein PAC-1and CD62p in progressive IS group were higher than the stable IS group.The difference was statistically significant (P<0.05).6. In our study, there were6patients were resistance to aspirin,105patients were sensitive to aspirin,5.4%of IS patients with AR.The expression of platelet membrane glycoprotein PAC-1、CD62p and CD63in AR group were significantly higher than in AS group,the difference was statistically significant(P<0.05).7. Platelet membrane GP Ⅱb HPA-3gene polymorphism is divided into aa,ab and bb. The expression of platelet membrane glycoprotein PAC-1and CD62p in patients who with bb genotype were higher than those with aa and ab genotype,but there’s no significant difference.Whether the platelet membrane glycoprotein in the people who suffered stroke with bb genotype were expressed higher.Further research is needed to confirm this view.8. The patients with GP Ⅱ b the HPA-3bb genotype were easier to suffer AR than those with aa and ab genotype,the difference was statistically significant (P=0.041).Suggesting bb genotype may be associated with aspirin resistance.Conclusion:1. The platelets were highly activated in patients with acute IS, so to detect the expression of platelet membrane glycoprotein PAC-1,CD62p,CD63by FCM can be used as a sensitive indicator of platelet activation.2. The expression of platelet membrane glycoprotein were higher in progressive IS group than in stable IS group.Prompted whether the platelet membrane glycoprotein expression were increased significantly in IS patients developed progressive IS possibility.3. The expression of platelet membrane glycoprotein and aggregation rate in high risk control group was higher than healthy people,suggesting the people who with stroke risk factors such as hypertension,coronary heart disease,diabetes,their platelet were activated.So we should control stroke risk factors in order to reduce the expression of platelet membrane glycoprotein and platelet aggregation rate,thereby reducing the incidence of stroke and thromboembolic events..4. Aspirin can reduce the expression of platelet membrane glycoprotein and aggregation rate,so aspirin can inhibit t platelet activation.5. The expression of platelet membrane glycoprotein PAC-1,CD62p and CD63in different GP Ⅱ b HPA-3genotypes was no significant difference, but the expression PAC-l,CD62p was higher in bb genotype than that of genotype aa and ab,the IS patients carrying bb genotype may be more likely to lead to high expression of platelet membrane glycoprotein. Further research is needed to confirm this view.6. The AR phenomenon does exist in IS, in this study,5.4%of IS patients with AR,meanwhile the platelet aggregation cannot be inhibited effectively after taking aspirin,and this phenomenon was high related with GPⅡb HPA-3bb genotype,suggesting platelet GPⅡb genotype of HPA-3bb gene may be an independent risk factor of AR. |
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