丹参饮对高脂血症模型大鼠血小板生理特性和功能的影响机制

Effects and mechanism of Danshen decoction on physiological characteristics and function of platelet in hyperlipidemia model rats

目的基于血小板膜糖蛋白4(CD36)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路探讨丹参饮对高脂血症模型大鼠血小板生理特性和功能的影响机制。方法通过喂养高脂饮食复制高脂血症模型大鼠,将其随机分为模型组、辛伐他汀组(0.004 g/kg)和丹参饮高、低剂量组(3.6、0.9 g/kg),另设空白组(喂养基础饲料),每组10只。各给药组大鼠每天灌胃相应药物,其余组灌胃等体积生理盐水,连续4周。末次给药后,检测大鼠血脂生化指标[总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)]含量,全血黏度、血浆黏度和血浆中纤维蛋白原(FIB)含量,血小板相关参数[血小板计数(PLT)、血小板分布宽度(PDW)和血小板平均体积(MPV)],血小板生理特性和功能相关因子[血管性血友病因子(vWF)、纤维连接蛋白(Fn)、磷脂酶A2(PLA2)、血栓素B2(TXB2)、6-酮-前列腺素F1α(6-keto-PGF1α)、血栓素A2(TXA2)、前列腺素(PGI2)、环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)、β-血小板球蛋白(β-TG)]水平,血小板聚集率(最大聚集率、60 s聚集率、180 s聚集率)和纤溶系统相关因子[组织型纤溶酶原活化剂(t-PA)、纤溶酶原激活物抑制剂1(PAI-1)]水平,血小板中CD36/PI3K/Akt信号通路相关蛋白[CD36、黏着斑激酶(FAK)、磷脂酰肌醇4-磷酸5-激酶(PIP5K)、PI3K、磷酸化Akt(p-Akt)、p-Akt1/2/3]表达水平。结果与空白组比较,模型组大鼠血浆中TC、TG、LDL-C含量,全血黏度、血浆黏度、血浆中FIB含量,血浆中PLT、MPV、vWF、Fn、PLA2、TXB2、TXA2、cGMP、β-TG水平,血小板最大聚集率、60 s聚集率,血浆中PAI-1水平,CD36、FAK、PIP5K、PI3K、p-Akt、p-Akt1/2/3蛋白表达水平均显著升高(P<0.05);HDL-C含量和6-keto-PGF1α、PGI2、t-PA水平均显著降低(P<0.05)。经丹参饮干预后,上述大部分指标均显著逆转(P<0.05)。结论丹参饮可改善高脂血症模型大鼠血小板生理特性和功能,其作用机制可能与下调CD36/PI3K/Akt信号通路活性、降低血小板活化程度有关。

OBJECTIVE To investigate the effects and mechanism of Danshen decoction influencing platelet physiological characteristics and function in hyperlipidemia model rats based on platelet membrane glycoprotein 4(CD36)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway.METHODS The hyperlipidemia model rats were induced by feeding highfat diet,and then randomly divided into model group,simvastatin group(0.004 g/kg),Danshen decoction high-dose and low-dose groups(3.6,0.9 g/kg),and blank group(fed with basic feed),with 10 rats in each group.Rats in each administration group were intragastrically administered with corresponding drugs every day,and the other groups were intragastrically administered with equal volume of normal saline for 4 weeks.After the last administration,the contents of blood lipid biochemical indexes[total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C)],whole blood viscosity,plasma viscosity and fibrinogen(FIB)content in plasma,platelet-related parameters[platelet count(PLT),platelet distribution width(PDW)and mean platelet volume(MPV)]were detected.The levels of platelet physiological characteristics and function-related factors[von Willebrand factor(vWF),fibronectin(Fn),phospholipase A2(PLA2),thromboxane B2(TXB2),6-keto-prostaglandin F1α(6-keto-PGF1α),thromboxane A2(TXA2),prostaglandin I2(PGI2),cyclic adenosine monophosphate(cAMP),cyclic guanosine monophosphate(cGMP),β-thromboglobulin(β-TG)],platelet aggregation rate(maximum aggregation rate,60 s aggregation rate,180 s aggregation rate)and fibrinolytic system-related factors[tissue plasminogen activator(t-PA),plasminogen activator inhibitor 1(PAI-1)]and the expressions of CD36/PI3K/Akt signaling pathway-related proteins[CD36,focal adhesion kinase(FAK),phosphatidylinositol 4-phosphate 5-kinase(PIP5K),PI3K,phosphorylated Akt(p-Akt),p-Akt1/2/3]in platelet were all determined.RESULTS Compared with blank group,the contents of TC,TG and LDL-C in plasma,whole blood viscosity,plasma viscosity,the plasma levels of FIB,PLT,MPV,vWF,Fn,PLA2,TXB2,TXA2,cGMP andβ-TG,maximum platelet aggregation rate,60 s aggregation rate,the expressions of PAI-1 in plasma,protein expressions of CD36,FAK,PIP5K,PI3K,p-Akt and p-Akt1/2/3 were significantly increased in the model group(P<0.05).The content of HDL-C and the levels of 6-keto-PGF1α,PGI2 and t-PA were significantly decreased(P<0.05).After the intervention of Danshen decoction,most of the above indexes were significantly reversed(P<0.05).CONCLUSIONS Danshen decoction can improve the physiological characteristics and function of platelets in hyperlipidemia model rats,and its mechanism may be related to the down-regulation of CD36/PI3K/Akt signaling pathway activity and the reduction of platelet activation.