The Regulation Of Nao Xin Tong To The Anti-platelet Effect Of Clopidogrel On The CYP2C19~*2 Gene Mutation In Volunteers With Qi Deficiency And Blood Stasis Constitution

Objective To investigate the effect of Nao xin tong on the platelet function of clopidogrel resistance groups with qi deficiency and blood stasis constitution related to CYP2C19*2 gene mutation.Methods 1. PCR-RFLP technology was used to genotype the CYP2C19*2 polymorphisms in 360 volunteers from 25-55 years old after physical examination. 2. Eighteen male volunteers with qi deficiency and blood stasis constitution ( 6 CYP2C19*1/*1, 6 CYP2C19*1/*2, 6 CYP2C19*2/*2 ) were enrolled in this study after signing informed consent. Each took clopidogrel 300mg on the first day and then 75mg once daily for consecutive six days . After seven washing days , NaoXinTong(0.8g,tid) was pretreated for five days. Then NaoXinTong combined clopidogrel were taken for 7 day(sthe dose was the same as above). Blood samples were obtained to measure the ADP-induced platelet aggregation by turbidimetry at baseline and 4 hours,24hours,3days and 7days after clopidogrel administration,and measure the platelet count, platelet functional and inflammatory index (ADP-induced platelet aggregation,CD62PandPAC-1 by flow cytometry,sCD40L by enzyme-linked Immunosorbnent assay) at baseline ,7 days after single clopidogrel administration, the seventh washing day,5 days after NaoXinTong administration and 7 days after clopidogrel combined NaoXinTong.Results 1. The frequencies of different genotypes of CYP2C19*2 were in agreement with Hardy-Weinberg equilibrium in our study(P>0.05).2.(1) At baseline there were no significant difference of BMI, blood biochemical indicators,platelet count,platelet aggregation, CD62P,PAC-1and sCD40L among the three different CYP2C19*2 genotypes. Nephelometry and flow cytometry determination of ADP-induced platelet aggregation has a good correlation(r=0.397,P=0.004).(2)There were significant decrease in ADP-induced platelet aggregation by turbidimetry at 4 hours,24hours,3days and 7days after clopidogrel administration among the three different CYP2C19*2 genotypes compared with that of the same genotype at baseline(P<0.01). ADP-induced platelet aggregation of CYP2C19*2/*2 genotype at 4 hours,24hours,3days and 7days were much higher than those of CYP2C19*1/*1 genotype (55.0士5.3%,52.1士2.4%,52.2士5.2% , 52.9士3.5%VS 22.2士3.8%,21.2士2.4%,23.1士4.5%,21.2士2.8% in CYP2C19*1/*1 in CYP2C19*2/*2, P<0.01).(3) There were significant decrease in ADP-induced platelet aggregation by flow cytometry after the administration of NaoXinTong among the three different CYP2C19*2 genotypes compared with that of the same genotype at the seventh washing day (P<0.01),but there were no significant difference among the three different CYP2C19*2 genotypes. (4) The platelet aggregation by flow cytometry ,CD62P,PAC-1 and sCD40L of CYP2C19*2/*2 genotype were significantly decreased at after two drugs'administration compared with the administration of NaoXinTong (2.1±0.2% VS 8.8±1.2%,22.2±1.1% VS 26.7±1.9%,20.4±1.2% VS 27.3±3.4%,1.2±0.3 VS 2.4±0.4, P <0.05).(5) The platelet aggregation by flow cytometry were significantly decreased in three different CYP2C19*2 genotypes (3.5±1.7% VS 7.2±1.5%,2.8±0.8% VS 16.1±2%, 2.1±0.2% VS 19.8±1.6%,P<0.01)between two drugs'administration and the administration of clopidogrel alone.Conclusions CYP2C19*2 genetic polymorphisms reduced to the inhibition of ADP-induced platelet aggregation by clopidogrel. The combination of clopidogrel with NaoXinTong can improve the curative effect of clopidogrel in the patients with CYP2C19 * 2 gene mutations .