Division of Urological Surgery, The Second Affiliated Hospital of Shantou University Medical College, Shantou, China.
Department of Clinical Laboratory Medicine, The Second Affiliated Hospital of Shantou University Medical College, Shantou, China.
Acta Haematol. 2021;144(5):551-559. doi: 10.1159/000514907. Epub 2021 Apr 13.
The objective of this study was to determine the expression of G protein-coupled receptors (GPCRs) in platelets from adult patients with chronic immune thrombocytopenic purpura (ITP).
Peripheral blood samples were collected from 40 patients with chronic ITP in the Second Affiliated Hospital of Shantou University Medical College, and 40 peripheral blood samples from healthy volunteers were collected; expressions of the adenosine diphosphate receptors (P2Y1 and P2Y12), alpha-2A adrenergic receptor (α2A-AR), and thromboxane A2 receptor (TP) in platelets were detected by flow cytometry. Gα protein, protease-activated receptor 1 (PAR1), and protease-activated receptor 4 (PAR4) were analyzed by Western blot and analyzed statistically.
Flow cytometry measurements of mean fluorescence intensities showed platelets from patients with chronic ITP, compared to healthy individuals, had significantly higher levels of P2Y1 (31.4 ± 2.2 vs. 7.8 ± 0.8), P2Y12 (29.6 ± 2.1 vs. 7.2 ± 1.3), α2A-AR (25.8 ± 2.9 vs. 9.8 ± 0.9), and TP (39.8 ± 3.1 vs. 4.7 ± 0.6) (all p < 0.01). Similarly, integrated optical density analysis of Western blots showed that platelets from patients with chronic ITP had significantly higher levels of Gα (1046.3 ± 159.96 vs. 254.49 ± 39.51), PAR1 (832.98 ± 98.81 vs. 203.92 ± 27.47), and PAR4 (1518.80 ± 272.45 vs. 431.27 ± 41.86) (all p < 0.01).
Expression of GPCRs is increased in platelets from patients with chronic ITP, suggesting that platelets of chronic ITP may participate in the complicated biological process by means of GPCR-mediated signaling pathways.
本研究旨在确定慢性免疫性血小板减少性紫癜（ITP）成年患者血小板中 G 蛋白偶联受体（GPCR）的表达。
收集汕头大学医学院第二附属医院 40 例慢性 ITP 患者外周血样本，收集 40 例健康志愿者外周血样本；用流式细胞术检测血小板中二磷酸腺苷受体（P2Y1 和 P2Y12）、α2A 肾上腺素能受体（α2A-AR）和血栓素 A2 受体（TP）的表达。用 Western blot 分析 Gα 蛋白、蛋白酶激活受体 1（PAR1）和蛋白酶激活受体 4（PAR4），并进行统计学分析。
流式细胞术测量平均荧光强度显示，与健康个体相比，慢性 ITP 患者的血小板中 P2Y1（31.4±2.2 比 7.8±0.8）、P2Y12（29.6±2.1 比 7.2±1.3）、α2A-AR（25.8±2.9 比 9.8±0.9）和 TP（39.8±3.1 比 4.7±0.6）水平显著升高（均 p<0.01）。同样，Western blot 分析的整合光密度显示，慢性 ITP 患者的血小板中 Gα（1046.3±159.96 比 254.49±39.51）、PAR1（832.98±98.81 比 203.92±27.47）和 PAR4（1518.80±272.45 比 431.27±41.86）水平显著升高（均 p<0.01）。
慢性 ITP 患者血小板中 GPCR 的表达增加，提示慢性 ITP 血小板可能通过 GPCR 介导的信号通路参与复杂的生物学过程。
