咪达唑仑对冠心病患者体外血小板活化反应的影响

目的 评价咪达唑仑对冠心病患者体外血小板活化反应的影响.方法 分别抽取10名健康志愿者及40例冠心病患者肱静脉血样5 ml,抗凝离心后得到富含血小板血浆(PRP).实验Ⅰ:每份PRP 1 ml,10份健康志愿者PRP作为正常对照组(ⅠC组,n=10),40份冠心病患者PRP随机分为4组(n:10):ⅠM0组、ⅠM1组、ⅠM2组和ⅠM3组Ⅰ.C组和ⅠM0组不加入咪达唑仑,ⅠM1组、ⅠM2组、ⅠM3组加入咪达唑仑,其终浓度分别为100、200、400ng/ml,孵育3 min,检测血小板聚集率.实验Ⅱ:将剩余PRP进一步离心后得到洗涤血小板(WP),每份WP 1 ml,10份健康志愿者WP作为正常对照组(ⅡC组),40份冠心病患者WP随机分为4组(n=10):ⅡM0组、ⅡM1组、ⅡM2组和ⅡM3组.ⅡC组和ⅡM0组不加入咪达唑仑,ⅡM1组、ⅡM2组、Ⅱ3组加入咪达唑仑,其终浓度分别为100、200、400 ng/ml,孵育3 min,采用竞争酶联免疫法检测血小板环磷酸腺苷(cAMP)和血栓烷B2(TXB2)的含量.结果 与ⅠC组比较,ⅠM0组血小板聚集率升高(P<0.01);与ⅠM1组比较,ⅠM2组和ⅠM3组血小板聚集率降低(P<0.01),ⅠM1组差异无统计学意义(P>0.05).与ⅡC组比较,ⅡM0组血小板cAMP含量降低、TXB2含量升高(P<0.01);与ⅠM0组比较,ⅡM1组和ⅡM3组血小板cAMP含量升高(P<0.05或0.01),,TXB2含量降低(P<0.01),ⅠM1组差异无统计学意义(P>0.05).结论 咪达唑仑200、400 ng/ml通过提高血小板cAMP水平,抑制其TXA2生成,可降低冠心病患者血小板的活化程度.

Effect of midazolam on platelet activation in patients with coronary heart disease in vitro

Objective To evaluate the effect of midazolam on platelet activation in patients with coronary heart disease (CHD) .Methods Brachial venous blood samples 5 ml drawn from 10 healthy adult volunteers and 40 patients with CHD were anticongulated with 3.8 % sodium citrate. Platelet rich plasma (PRP) 3 ml was obtained by centrifngation at 800 rpm for 8 min. Experiment Ⅰ : Ten portions of PRP from healthy adult volunteers served as control group with 1 ml for each (group ⅠC, n = 10). Forty portions of PRP from patients with CHD were randomly divided into 4 groups (n = 10 each): ⅠM0, ⅠM1,ⅠM2 and ⅠM3. In group ⅠC and ⅠM0, midazolam was not added while in group ⅠM1 , ⅠM2 and ⅠM3, midazolam was added with the final concentration at 100, 200 and 400 ng/ml respectively and the PRPs were then incubated for 3 min. The platelet aggregation rote was determined using turbidimetric method. Experiment Ⅱ : After the remaining PRP was eentrifnged at 2000 r/ rain for 5 min, washed platelets (WPs) were obtained. Ten portions of WPs from healthy adult volunteers served as control group with 1 ml for each (group ⅡC, n = 10). Forty portions of WPs from patients with CHD were randomly divided into 4 groups (n = 10 each) : ⅡM0, ⅡM1, ⅡM2 and ⅡM3· In group ⅡC and ⅡM0, midazolam was not added while in group Ⅱ, ⅡM2 and ⅡM3, midazolam was added with the final concentration at 100, 200 and 400 ng/ml respectively and the WPs were then incubated for 3 min. The content of platelet cAMP and were measured by enzyme immunoassay. Results Platelet aggregation rate was significantly higher in group ⅠM0 than in group ⅠC(P<0.01). Compared with groupⅠM0, platelet aggregation rate was significantly decreased in group ⅠM2 and ⅠM3(P<0.01) but there was no significant change in group ⅠM1 (P >0.05). The content of platelet cAMP was significantly decreased while the level of TXB2 was significantly increased in group ⅡM0 as compared with group ⅡC (P < 0.01). Compared with group Ⅱ M0, the content of platelet cAMP was significantly increased (P <0.05 or 0.01) while the level of TXB2 was significantly decreased in group ⅡM2 and ⅡM3(P< 0.01), but there was no significant change in group ⅡM1 (P > 0.05). Conclusion Midazolam 200 and 400 ng/ml can inhibit the platelet activation through increasing the content of platelet cAMP and reducing the production of TXA2 in vitro.