益气活血方联合PI3K抑制剂Wortmannin对血小板聚集率的影响  

Effects of Yiqi Huoxue Granule Combined with PI3K Inhibitor on Platelet Aggregation Rate

作　　者：吴鸿[1] 雷震[1] 高水波[1] 韩勇军[1] 王振涛[1] 韩丽华[1] WU Hong;LEI Zhen;GAO Shuibo;HAN Yongjun;WANG Zhentao;HAN Lihua(Henan University of Chinese Medicine,Zhengzhou Henan China 450046)

机构地区：[1]河南中医药大学,河南郑州450046

出　　处：《中医学报》2018年第11期2151-2155,共5页Acta Chinese Medicine

基　　金：国家自然科学基金项目(81072968);教育部留学回国人员科研启动基金项目[教外司(2011)1568];河南省重点科技攻关项目(102102310334)

摘　　要：目的:观察益气活血方联合PI3K抑制剂Wortmannin对大鼠体外血小板聚集率的影响。方法:高剂量、中剂量、低剂量益气活血方(0. 2 g·L^(-1)、0. 1 g·L^(-1)、0. 05 g·L^(-1))体外干预血小板,用比浊法检测二磷酸腺苷(adenosine diphosphate,ADP)诱导的血小板聚集水平,用钙离子探针fura-2/AM观察血小板胞内Ca2+变化,用q PCR和Western blot法分别检测钙池调控Ca2+流入通道(store-operated calcium channel,SOCC)成分Orai1和基质交感分子1(stromal interaction molecule l,STIM1) mRNA及蛋白表达水平。结果:与对照组比较,益气活血方中剂量组(0. 1 g·L^(-1))和高剂量组(0. 2 g·L^(-1))血小板聚集率显著降低(P <0. 05),益气活血方高剂量组(0. 2 g·L^(-1))血小板胞质内Ca2+浓度显著降低(P <0. 05)。益气活血方高剂量组(0. 2 g·L^(-1)) SOCC通路上STIM1、Orai1的mRNA及蛋白表达水平显著降低(P <0. 01);联合应用PI3K抑制剂Wortmannin能显著抑制血小板聚集率(P <0. 01),抑制作用明显好于单独应用中药或西药。结论:益气活血方联合应用PI3K抑制剂Wortmannin能显著抑制血小板聚集率,其作用与抑制STIM1/Orai1活性有关。Objective: To observe the effects of Yiqi Huoxue Granule combined with PI3K inhibitor Wortmannin on platelet aggregation rate in vitro. Methods: High-dose,medium-dose,and low-dose of Yiqi Huoxue Granule ( 0. 2 g·L^-1,0 . 1 g·L^-1,0 . 05 g· L^-1 ) was used to intervene platelets in vitro,and the platelet aggregation level induced by adenosine diphosphate ( ADP) was detected by turbidimetry. Calcium ion probe fura-2 /AM was used to observe intracellular Ca2 + changes in platelets,and qPCR and Western blot were used to detect the store-operated calcium channel ( SOCC) component Orai1 and stromal interaction molecule 1 ( STIM1) mRNA and protein expression levels. Results: Compared with the control group,the platelet aggregation rate in middle dose group( 0. 1 g·L^-1 ) and high dose group ( 0. 2 g·L^-1 ) of Yiqi Huoxue Granule was significantly lower ( P < 0. 05) ,platelet cytoplasmic Ca2 + concentration in Yiqi Huoxue Granule high dose Group ( 0. 2 g·L^-1 ) was significantly decreased ( P < 0. 05) . The mRNA and protein expression levels of STIM1 and Orai1 in the high-dose group ( 0. 2 g·L^-1 ) of Yiqi Huoxue Granule were significantly decreased ( P < 0. 01) . Combined use of Yiqi Huoxue Granule with PI3K inhibitor Wortmannin significantly inhibited platelet aggregation rate ( P < 0. 01) ,and the inhibitory effect was significantly better than that of Chinese medicine or western medicine alone. Conclusion: Yiqi Huoxue Granule in combination with PI3K inhibitors can inhibit platelet aggregation rate,which is related to inhibition of STIM1 /Orai1 activity.

关 键 词：益气活血方 血小板聚集率 STIM1 Orai1 PI3K抑制剂 大鼠 

分 类 号：R285.5[医药卫生—中药学]