龙柴降血方对原发性血小板增多症患者凝血功能及血小板活化蛋白的影响

Effects of Longchai decoction for regulating platelet on coagulation function and platelet activating protein in patients with essential thrombocythemia

目的观察龙柴降血方对原发性血小板增多症(ET)患者凝血功能及血小板活化蛋白的影响。方法选取2020年1月—2021年10月中国中医科学院西苑医院收治的ET患者22例,给予龙柴降血方治疗4个月,观察治疗前后血栓弹力图指标[凝血时间(R)、凝血速率(K)、纤维蛋白功能(Angle)、血栓最大振幅(MA)、血栓最大振幅时间(TMA)、即时振幅(A)、30 min振幅(A30)、从血样开始检测到曲线分叉所需时间(SP)、血凝块溶解时间(CLT)]、高凝状态发生率和积分变化。选取其中15例ET患者,采用平行反应监测(PRM)技术检测患者治疗前后血小板活化蛋白[血小板膜糖蛋白Ibα(GPIbα)、糖蛋白VI(GPVI)、选择素-P(SELP)]表达水平,分析比较血小板聚集改善患者和血小板聚集增强患者上述各蛋白表达水平的差异;取5例健康者和10例ET患者治疗前后血清,采用Western blot法检测血清GPIbα表达情况;分析15例ET患者MA与血清GPIbα表达水平的相关性。结果与治疗前比较,龙柴降血方治疗后MA、CLT、高凝状态发生率、高凝状态积分和血清GPIbα、GPVI表达水平均明显降低(P均<0.05),血清SELP表达水平无明显变化(P>0.05)。治疗后血小板聚集改善患者血清GPIbα表达水平明显低于血小板聚集增强患者(P<0.05),GPVI表达水平无明显变化(P>0.05)。与健康人相比,10例ET患者治疗前血清GPIbα相对表达量明显增高(P<0.05);10例ET患者治疗后血清GPIbα相对表达量较治疗前明显降低(P<0.05)。龙柴降血方治疗后MA下降与GPIbα表达下调呈正相关(r=0.6986,P<0.05)。结论龙柴降血方能够抑制ET患者血小板聚集,改善高凝状态,其机制可能与抑制GPIbα蛋白介导的血小板活化信号通路有关。

Objective It is to observe the effects of Longchai decoction for regulating platelet on coagulation function and platelet activating protein in patients with essential thrombocythemia(ET).Methods Twenty-two ET patients treated in Xiyuan Hospital of China Academy of Chinese Medical Science from January 2020 to October 2021 were selected and treated with Longchai decoction for regulating platelet for 4 months,the indexes of thromboela-stogram[reaction time(R),kinetics time(K),kinetics of clot development(Angle),maximal amplitude(MA),time to MA(TMA),amplitude(A),30 min amplitude(A30),SP,clot lysis time(CLT)],incidence and scores of hypercoagulable state before and after the treatment were observed.Fifteen of the ET patients were selected,and their expression levels of platelet activation proteins[glycoprotein Ibα(GPIbα),glycoprotein VI(GPVI),and selectin-P(SELP)]before and after treatment were detected by the parallel response monitoring(PRM)technique to analyze and compare the differences in the expression levels of the above proteins between the patients with improved platelet aggregation and the patients with enhanced platelet aggregation;the serum of 5 healthy patients and 10 ET patients were taken before and after treatment to detect the expression of serum GPIbαby Western blot method;the correlation between MA and the expression level of serum GPIbαin 15 ET patients was analyzed.Results Compared with before treatment,the MA,CLT,incidence and scores of hypercoagulable state,and expression levels of serum GPIbαand GPVI were significantly reduced after treatment with Longchai decoction for regulating platelet(all P<0.05),and there was no significant difference in the expression level of serum SELP(P>0.05).The expression level of serum GPIbαwas significantly lower in patients with improved platelet aggregation than that in patients with enhanced platelet aggregation after treatment(P<0.05),but there was no significant change in the expression level of GPVI(P>0.05).Compared with healthy people,the relative expression of serum GPIbαwas significantly higher in 10 ET patients before treatment(P<0.05);the relative expression of serum GPIbαwas significantly decreased in 10 ET patients after treatment compared with before treatment(P<0.05).There was a positive correlation between the decrease of MA and the down-regulation of GPIbαexpression after treatment with Longchai decoction for regulating platelet(r=0.6986,P<0.05).Conclusion Longchai decoction for regulating platelet can inhibit platelet aggregation and improve hypercoagulability in patients with ET,and its mechanism may be related to the inhibition of GPIbαprotein-mediated platelet activation signaling pathway.