基于蛋白质组学探讨丹参饮对高脂血症模型大鼠血小板活化的影响机制  

Mechanism of Danshenyin in Inhibiting Platelet Activation in Rat Model of Hyperlipidemia Based on Proteomics

作　　者：张玉昆 冯月男 卞敬琦 刘欣欣 肖洪彬 牛雯颖 ZHANG Yukun;FENG Yuenan;BIAN Jingqi;LIU Xinxin;XIAO Hongbin;NIU Wenying(Key Laboratory of Basic and Applied Research on Northern Medicine,Ministry of Education,Experimental Teaching and Training Center,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学实验实训中心,黑龙江中医药大学教育部北药基础与应用研究重点实验室,哈尔滨150040

出　　处：《中国实验方剂学杂志》2024年第8期1-8,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82074326);黑龙江省博士后面上项目(LBH-Z20195)。

摘　　要：目的:通过长期高脂饲料喂养复制高脂血症动物模型,采用蛋白质组学、分子生物学等方法阐释丹参饮对高脂血症模型大鼠血小板活化的影响机制。方法:Wistar大鼠,按血脂水平随机分正常组、模型组、丹参饮组,每组10只。正常组动物喂食基础饲料,模型组和丹参饮组动物喂食高脂饲料造模,3组动物均自由饮水、自由进食。于第9周开始治疗干预,丹参饮组实验动物给予丹参饮灌胃给药,剂量为3.6 g·kg^(-1)生药;模型组和正常组实验动物根据体质量给予等体积生理盐水。第12周后取材并进行相关指标检测,利用生化分析仪来检测血清中血脂4项水平,利用血液流变仪来检测全血黏度和血浆黏度,利用液质联用色谱仪来检测血小板蛋白质组,蛋白免疫印迹法(Western blot)检测血小板膜糖蛋白4(CD36)、黏着斑激酶(FAK)、磷脂酰肌醇4-磷酸5-激酶(PIP5K)、磷酸化磷脂酰肌醇3-激酶(p-PI3K)、蛋白激酶(Akt)、磷酸化蛋白激酶B(p-Akt)蛋白表达。结果:与模型组比较,丹参饮组高脂血症模型大鼠血浆中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)水平能够明显降低(P<0.05),高密度脂蛋白胆固醇(HDL-C)水平明显升高(P<0.05);与模型组比较,丹参饮组大鼠血小板聚集率明显降低(P<0.05);模型组与正常组比较上调蛋白数目44个,下调蛋白数目12个,差异蛋白总数为56个;丹参饮组与模型组比较上调蛋白数目21个,下调蛋白数目22个,差异蛋白总数为43个;丹参饮组和正常组比较上调蛋白数目31个,下调蛋白数目49个差异蛋白总数共计80个,通过基因本体(GO)功能富集分析主要涉及胆固醇转运和外排,以及细胞因子的产生比较显著且与血脂异常和血小板活化有关,京都基因与基因组百科全书(KEGG)功能富集分析涉及ECM-receptor interaction信号通路、过氧化物酶体增殖物激活受体(PPAR)信号通路、黏着斑信号通路、PI3K/AObjective:To decipher the mechanism of Danshenyin in regulating platelet activation in the rat model of hyperlipidemia by means of proteomics and molecular biology.Method:Wistar rats were randomized into blank,model,and Danshenyin groups(n=10)according to the blood lipid level.The rats in the blank group were fed with a basic diet,and those in the model and Danshenyin groups with a high-fat diet.All the rats had free access to water and food.The treatment began at the 9th week.The rats in the Danshenyin group were administrated with Danshenyin by gavage at a crude drug dose of 3.6 g·kg^(-1).The rats in the model and blank groups were administrated with an equal volume of normal saline according to body weight.At the 12th week,the tissue samples were collected for the measurement of related indicators,and the blood lipid level was measured by an automatic biochemical analyzer.The whole blood viscosity and plasma viscosity were measured by an automatic hemorheometer.The platelet proteome was determined by liquid chromatography-mass spectrometry.Western blotting was employed to determine the protein levels of platelet membrane glycoprotein 4(CD36),focal adhesion kinase(FAK),phosphatidylinositol 4-phosphate 5-kinase(PIP5K),phosphorylated phosphatidylinositol 3-kinase(p-PI3K),protein kinase B(Akt),and phosphorylated protein kinase B(p-Akt).Result:Compared with the model group,Danshenyin lowered the levels of total cholesterol(TC),triglyceride(TG),and low-density lipoprotein cholesterol(LDL-C)in the plasma(P<0.05),elevated the level of highdensity lipoprotein cholesterol(HDL-C)(P<0.05),and reduced the platelet aggregation rate(P<0.05).Compared with the blank group,the modeling up-regulated the expression of 44 proteins and down-regulated the expression of 12 proteins.Compared with the model group,Danshenyin up-regulated the expression of 21 proteins and down-regulated the expression of 22 proteins.Compared with the blank group,Danshenyin upregulated the expression of 31 proteins and down-regulated the expression of 49

关 键 词：高脂血症 丹参饮 活血化瘀 血小板膜糖蛋白4 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学] R289R287R22R2-031R33R24