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目的 :观察成人慢性免疫性血小板减少性紫癜(immune thrombocytopenic purpura,ITP)患者血小板G蛋白偶联受体的变化。方法:采用流式细胞仪检测ITP患者血小板表达二磷酸腺苷(adenosine diphosphate,ADP)受体(P2Y1、P2Y12)、α2A-肾上腺素能受体(α2A-adrenergic receptor,α2A-AR)和血栓烷A2受体(thromboxane A2receptor,TXA2-R)的表达,采用蛋白印迹法检测血小板Gα蛋白、蛋白酶激活受体-1(proteinase-activated receptor 1,PAR-1)和蛋白酶激活受体-4(proteinase-activated receptor 4,PAR-4)的表达。结果:慢性ITP患者血小板表达P2Y1、P2Y12、α2AAR和TXA2-R的平均荧光强度分别为31.4±2.2、29.6±2.1、25.8±2.9和39.8±3.1,明显高于健康对照者(分别为7.8±0.8、7.2±1.3、9.8±0.9和4.7±0.6),差异有统计学意义(P<0.01)。蛋白印迹法检测显示,慢性ITP患者血小板表达Gα蛋白、PAR-1和PAR-4的累积光密度值分别为1 046.3±159.96、832.98±98.81和1 518.80±272.45,也显著高于健康对照者(分别为254.49±39.51、203.92±27.47和431.27±41.86),差异有统计学意义(P<0.01)。结论:成人慢性ITP患者血小板G蛋白偶联受体的表达增多,提示慢性ITP患者的血小板可能通过G蛋白偶联受体的信号转导途径,参与了复杂的生物学过程。
Objective: To observe the changes of platelet G protein-coupled receptors in adult patients with chronic thrombocytopenic purpura (ITP). Methods: Flow cytometry was used to detect the expression of platelet-derived adenosine diphosphate (ADP) receptors (P2Y1 and P2Y12), α2A-adrenergic receptor (α2A-AR) and thromboxane A2 receptor (TXA2-R), the expression of platelet Gα protein, proteinase-activated receptor 1 (PAR-1) and proteinase- activated receptor 4, PAR-4). Results: The average fluorescence intensity of platelets expressing P2Y1, P2Y12, α2AAR and TXA2-R in patients with chronic ITP were 31.4 ± 2.2, 29.6 ± 2.1, 25.8 ± 2.9 and 39.8 ± 3.1, respectively, which were significantly higher than those in healthy controls (7.8 ± 0.8 , 7.2 ± 1.3, 9.8 ± 0.9 and 4.7 ± 0.6 respectively), the difference was statistically significant (P <0.01). Western blotting showed that the cumulative optical density of Gα protein expression in platelet of patients with chronic ITP was 1 046.3 ± 159.96, 832.98 ± 98.81 and 1 518.80 ± 272.45 respectively, which was also significantly higher than that of healthy controls ( Respectively, 254.49 ± 39.51,203.92 ± 27.47 and 431.27 ± 41.86), the difference was statistically significant (P <0.01). Conclusion: The expression of platelet G protein-coupled receptor in adult chronic ITP patients is increased, suggesting that platelets in chronic ITP patients may participate in complex biological processes through the signal transduction pathway of G protein-coupled receptors.