| Objection:The aim of this study was to investigate the relationship between the expression level of plasma micro RNA-223 and platelet reactivity in patients with acute ischemic stroke(AIS),to evaluate its predictive value in clopidogrel high on-treatment platelet reactivity(HTPR),and explore the relationship between plasma micro RNA-223 expression levels and CYP2C19 gene polymorphism.Methods:During the period from November 2017 to November 2018,patients in the control group and AIS group who were hospitalized in the Department of Neurology,Second Hospital of Tianjin Medical University,recorded clinical data and laboratory tests and results.Flow cytometry was used to detect the phosphorylation level of vasodilator stimulated phosphoprotein(VASP)and ADP-induced platelet aggregation(ADP-PAg)in platelets,method to assess platelet reactivity.and real-time PCR was used to detect the expression level of plasma micro RNA-223.The CYP2C19loss-of-function(LOF)allele was typed by polymerase chain reaction-restriction fragment length polymorphism(common polymorphisms *2(G681A)and *3(G636A))).The control group was tested for platelet aggregation rate by VASP and the expression level of plasma micro RNA-223 was measured at admission.Patients in the AIS group were tested for platelet aggregation before and after 7±1 days of clopidogrel treatment using both ADP-PAg and VASP methods.Then the relative inhibition rate(Relative Inhibition,RI)calculated according to the ADP-PAg method is grouped in quantiles.The upper quartile was clopidogrel high-response group(clopidogrel non-HTPR group)and lower quad The number of digits is clopidogrel low-response group(clopidogrel HTPR group).Finally,the detection of plasma micro RNA-223 expression level and CYP2C19 genotype determination before and after treatment in patients with AIS.The clinical outcome of the patients was assessed by a modified Rankin score at the end of March.Correlation between plasma micro RNA-223 expression level and clopidogrel HTPR in patients with AIS by unconditional logistic regression.The predictive value of plasma microrna-223 expression level for the HTPR of clopidogrel in AIS patients and the adverse clinical outcomes in AIS patients was analyzed using the subject operating characteristic curve.Results:(1)In 120 patients with platelet aggregation rate detected by ADP-PAg and VASP,VASP-PRI was positively correlated with ADP-PAg before and after treatment(Before administration: r=0.190,P=0.037;after administration: r=0.323,P<0.001)(2)Plasma micro RNA-223 level and VASP-PRI level in the AIS group before treatment were higher than those in the control group(before administration mi R-223: 1.254 ± 0.80vs1.001 ± 0.51,P=0.041;before administration VASP-PRI:66.23 ± 11.32 vs 34.77 ± 10.69,P<0.001).Logistic regression analysis showed that elevated VASP-PRI before administration was associated with increased risk of AIS(OR 1.011,95% CI 1.004-1.019,P=0.002)(3)Among the 60 AIS patients screened by the quarterwise method of RI,the expression level of microrna-223 after treatment was negatively correlated with VASP-PRI after treatment(r=-0.304,P=0.018).(4)Plasma micro RNA-223 levels before and after clopidogrel treatment in patients with AIS were lower than those in non-HTPR group after clopidogrel treatment(before administration mi R-223: 1.033±0.910 vs 1.575±0.536,P=0.032;after administration mi R-223: 0.521±0.379 vs 0.676±0.328,P=0.034).Logistic regression analysis showed that the decrease of plasma micro RNA-223 expression level before and after treatment was associated with an increased risk of clopidogrel HTPR in AIS patients(pre-treatment mi R-223: OR 2.219,95% CI 1.030-4.781,p=0.042;after administration mi R-223:OR 5.109,95% CI 1.082-24.133,p=0.039).The results of ROC curve showed that the expression level of plasma micro RNA-223 was less than1.185 before AIS patients were prone to clopidogrel HTPR.The plasma micro RNA-223 expression level was analyzed by a grouping of 1.185,and it was found that the micro RNA-223 expression level decreased(<1.185)CYP2C19 LOF allele carrying number compared with the micro RNA-223 expression level(?1.185)group.More,but the difference did not reach statistical difference(P>0.05)(5)By interaction analysis of unconditional logistic regression,there was an interaction between pretreatment plasma micro RNA-223 expression level and whether or not the CYP2C19 LOF allele was carried(OR 3.750,95% CI 1.078-13.039,p=0.038),plasma micro RNA after administration.There was no interaction between the expression level of-223 and whether or not the CYP2C19 LOF allele was carried(OR 1.266,95% CI 0.297-5.399,p=0.750).The CYP2C19 LOF allele-bearing status group was found to be in the CYP2C19 LOF allele carrier.The expression level of plasma micro RNA-223 in HTPR group was lower than that in non-HTPR group after clopidogrel treatment,and it was statistically significant(P <0.05).In the non-carriers of CYP2C19 LOF allele,the expression level of plasma micro RNA-223 had no significant relationship with clopidogrel HTPR(P>0.05).(6)Decreased expression of plasma micro RNA-223 before and after treatment was associated with an increased risk of clinically poor outcome(pre-treatment mi R-223: OR 1.929,95% CI 1.250-2.976,P= 0.003;after administrationmi R-223: OR 3.079,95%CI 1.360-6.969,P =0.007).The results of ROC curve showed that the decrease of plasma micro RNA-223 expression level in patients with AIS can predict the poor clinical outcome.Less than 1.148 is prone to poor outcome(optimum cutoff value 1.148,area under the curve is 0.878),and plasma micro RNA-223 expression level below 0.633 is prone to poor outcome(optimum cutoff value 0.633,area under the curve is 0.776).Conclusion: An increase in platelet aggregation rate before administration increases the risk of AIS.Decreased plasma micro RNA-223 expression levels before and after treatment increased the risk of clopidogrel HTPR in AIS patients,and the decrease in plasma micro RNA-223 expression levels before and after treatment was associated with an increased risk of clinical outcomes.The CYP2C19 LOF allele interacts with the expression level of plasma micro RNA-223,and the CYP2C19 LOF allele carrying may make the differential expression of plasma micro RNA-223 more distinct.The study suggests that combined detection of plasma micro RNA-223 and CYP2C19 gene polymorphism may be more effective in predicting the occurrence of clopidogrel HTPR in patients with AIS,and provide an objective basis for future studies of individualized differences in clinical antiplatelet drugs. |
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