Objective： To investigate a possible association between common variations of the P2RY12 and the residual clopidogrel on-treatment platelet reactivity after adjusting for the influence of CYP2C19 tested by thromboe- ...Objective： To investigate a possible association between common variations of the P2RY12 and the residual clopidogrel on-treatment platelet reactivity after adjusting for the influence of CYP2C19 tested by thromboe- lastography （TEG）. Methods： One hundred and eighty patients with acute coronary syndrome （ACS） treated with clopJdogrel and aspJdn were included and platelet function was assessed by TEG. Five selected P2RY12 single nu- cleotide polymorphisms （SNPs; rs6798347, rs6787801, rs6801273, rs6785930, and rs2046934）, which cover the common variations in the P2RY12 gene and its regulatory regions, and three CYP2C19 SNPs （ 2, 3, 17） were geno- typed and possible haplotypes were analyzed. Results： The high on-treatment platelet reactivity （HTPR） prevalence defined by a platelet inhibition rate 〈30% by TEG in adenosine diphosphate （ADP）-channel was 69 （38.33%）. Six common haplotypes were inferred from four of the selected P2RY12 SNPs （denoted H0 to H5） according to the linkage disequilibrium R square （except for rs2046934）. Haplotype H1 showed a significantly lower incidence of HTPR than the reference haplotype （H0） in the total study population while haplotypes H1 and H2 showed significantly lower incidences of HTPR than H0 in the nonsmoker subgroup after adjusting for CYP2Clg effects and demographic characteristics. rs2046934 （T744C） did not show any significant association with HTPR. Conclusions： The combination of common P2RY12 variations including regulatory regions rather than rs2046934 （T744C） that related to pharmacodynamics of clopidogrel in patients with ACS was independently associated with residual on-clopidogrel platelet reactivity. This is apart from the established association of the CYP2C19. This association seemed more important in the subgroup defined by smoking.展开更多
该研究基于多源异构生物医学数据整合分析,并结合动物实验验证,系统探讨健脾活骨方治疗激素性股骨头坏死(SONFH)的优势作用环节及其分子机制。研究首先通过中医药百科全书(ETCM v 2.0)获取健脾活骨方所含候选活性成分及靶点;同时借助HP...该研究基于多源异构生物医学数据整合分析,并结合动物实验验证,系统探讨健脾活骨方治疗激素性股骨头坏死(SONFH)的优势作用环节及其分子机制。研究首先通过中医药百科全书(ETCM v 2.0)获取健脾活骨方所含候选活性成分及靶点;同时借助HPO数据库挖掘该证候的潜在关联基因;最后,运用临床转录组学检测获取SONFH痰瘀阻络证的相关靶点。对上述三类靶点取交集后,构建“中药-成分-疾病-证候”多维网络。利用STRING数据库进行蛋白质-蛋白质相互作用(PPI)网络分析,并通过DAVID平台对核心靶点进行京都基因与基因组百科全书(KEGG)通路富集分析,以预测其关键生物学过程与信号通路。最后,通过制备SONFH痰瘀阻络证大鼠模型进行药效学和机制验证。经数据整合挖掘获得包含146个网络核心靶标的“病证基因-方药靶标”网络,通路富集分析表明,健脾活骨方主要通过嘌呤能受体P2Y12(P2RY12)-磷脂酰肌醇3-激酶(PI3K)-丝氨酸/苏氨酸激酶(Akt)-纤维蛋白原γ链(FGG)信号通路,调节血脂偶联干预SONFH痰瘀阻络表型。HE染色结果发现,SONFH痰瘀阻络证大鼠的股骨头组织钙化比列降低,骨小梁排列松散,骨细胞欠饱满,空骨陷窝增多明显,骨髓内脂肪细胞增多;血脂四项检测表明,SONFH痰瘀阻络证大鼠甘油三酯、总胆固醇、低密度脂蛋白升高,高密度脂蛋白降低;凝血四项检测表明,SONFH痰瘀阻络证大鼠活化部分凝血活酶时间、凝血酶原时间降低,凝血酶时间和血浆纤维蛋白原升高。免疫组织化学染色结果显示,与正常组比较,模型组大鼠股骨头组织中P2RY12、磷酸化PI3K(p-PI3K)、磷酸化Akt(p-Akt)和FGG蛋白的阳性表达面积显著增加(P<0.05);而各给药组上述蛋白表达均显著降低(P<0.05),且高剂量组抑制效果最为明显,表明健脾活骨方能够剂量依赖性地调控P2RY12-PI3K-Akt-FGG信号轴。综上所述,健脾活骨方可通过多靶点、多通路协同作用,尤其是抑制P2RY12-PI3K-Akt-FGG信号轴的过度激活,改善SONFH痰瘀阻络证的血脂及骨代谢异常,发挥对证治疗作用。展开更多
基金Project supported by the Beijing Higher Education Young Elite Teacher Project(No.YETP0064),China
文摘Objective： To investigate a possible association between common variations of the P2RY12 and the residual clopidogrel on-treatment platelet reactivity after adjusting for the influence of CYP2C19 tested by thromboe- lastography （TEG）. Methods： One hundred and eighty patients with acute coronary syndrome （ACS） treated with clopJdogrel and aspJdn were included and platelet function was assessed by TEG. Five selected P2RY12 single nu- cleotide polymorphisms （SNPs; rs6798347, rs6787801, rs6801273, rs6785930, and rs2046934）, which cover the common variations in the P2RY12 gene and its regulatory regions, and three CYP2C19 SNPs （ 2, 3, 17） were geno- typed and possible haplotypes were analyzed. Results： The high on-treatment platelet reactivity （HTPR） prevalence defined by a platelet inhibition rate 〈30% by TEG in adenosine diphosphate （ADP）-channel was 69 （38.33%）. Six common haplotypes were inferred from four of the selected P2RY12 SNPs （denoted H0 to H5） according to the linkage disequilibrium R square （except for rs2046934）. Haplotype H1 showed a significantly lower incidence of HTPR than the reference haplotype （H0） in the total study population while haplotypes H1 and H2 showed significantly lower incidences of HTPR than H0 in the nonsmoker subgroup after adjusting for CYP2Clg effects and demographic characteristics. rs2046934 （T744C） did not show any significant association with HTPR. Conclusions： The combination of common P2RY12 variations including regulatory regions rather than rs2046934 （T744C） that related to pharmacodynamics of clopidogrel in patients with ACS was independently associated with residual on-clopidogrel platelet reactivity. This is apart from the established association of the CYP2C19. This association seemed more important in the subgroup defined by smoking.
文摘该研究基于多源异构生物医学数据整合分析,并结合动物实验验证,系统探讨健脾活骨方治疗激素性股骨头坏死(SONFH)的优势作用环节及其分子机制。研究首先通过中医药百科全书(ETCM v 2.0)获取健脾活骨方所含候选活性成分及靶点;同时借助HPO数据库挖掘该证候的潜在关联基因;最后,运用临床转录组学检测获取SONFH痰瘀阻络证的相关靶点。对上述三类靶点取交集后,构建“中药-成分-疾病-证候”多维网络。利用STRING数据库进行蛋白质-蛋白质相互作用(PPI)网络分析,并通过DAVID平台对核心靶点进行京都基因与基因组百科全书(KEGG)通路富集分析,以预测其关键生物学过程与信号通路。最后,通过制备SONFH痰瘀阻络证大鼠模型进行药效学和机制验证。经数据整合挖掘获得包含146个网络核心靶标的“病证基因-方药靶标”网络,通路富集分析表明,健脾活骨方主要通过嘌呤能受体P2Y12(P2RY12)-磷脂酰肌醇3-激酶(PI3K)-丝氨酸/苏氨酸激酶(Akt)-纤维蛋白原γ链(FGG)信号通路,调节血脂偶联干预SONFH痰瘀阻络表型。HE染色结果发现,SONFH痰瘀阻络证大鼠的股骨头组织钙化比列降低,骨小梁排列松散,骨细胞欠饱满,空骨陷窝增多明显,骨髓内脂肪细胞增多;血脂四项检测表明,SONFH痰瘀阻络证大鼠甘油三酯、总胆固醇、低密度脂蛋白升高,高密度脂蛋白降低;凝血四项检测表明,SONFH痰瘀阻络证大鼠活化部分凝血活酶时间、凝血酶原时间降低,凝血酶时间和血浆纤维蛋白原升高。免疫组织化学染色结果显示,与正常组比较,模型组大鼠股骨头组织中P2RY12、磷酸化PI3K(p-PI3K)、磷酸化Akt(p-Akt)和FGG蛋白的阳性表达面积显著增加(P<0.05);而各给药组上述蛋白表达均显著降低(P<0.05),且高剂量组抑制效果最为明显,表明健脾活骨方能够剂量依赖性地调控P2RY12-PI3K-Akt-FGG信号轴。综上所述,健脾活骨方可通过多靶点、多通路协同作用,尤其是抑制P2RY12-PI3K-Akt-FGG信号轴的过度激活,改善SONFH痰瘀阻络证的血脂及骨代谢异常,发挥对证治疗作用。
