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成人慢性免疫性血小板减少性紫癜患者血小板中钙网蛋白作用的研究

Study on the Role of Calreticulin Within Platelet from Adult Patients with Chronic Immune Thrombocytopenic Purpura.

作者信息

Yung Kam Chau, Zhang Ze Wen, Yu Wen Jun, Qiu Jin Feng, Xu Cheng Wei, He Chun Ling, Xu Xian Ru, Yin Jun

机构信息

1Division of Hematology, The Second Affiliated Hospital of Shantou University Medical College, Dongxia Road North, Shantou, 515041 Guangdong Province China.

3Division of Respiration, The Second Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong Province China.

出版信息

Indian J Hematol Blood Transfus. 2018 Oct;34(4):711-718. doi: 10.1007/s12288-018-0955-8. Epub 2018 Apr 2.

Abstract

To observe the differences in proteins between adult patients with chronic immune thrombocytopenic purpura (ITP) and healthy adults. 30 patients with chronic ITP and 30 healthy controls were enrolled into the study. The platelet total protein was extracted from peripheral venous blood of 10 chronic ITP patients and 10 healthy controls respectively, and subjected to two-dimensional electrophoresis (2-DE) to find the differential protein spot between chronic ITP patients and healthy controls, then the differential protein spots were identified by mass spectrometry. Subsequently, platelets RNA and proteins were isolated from the other 20 chronic ITP patients and 20 healthy controls respectively, and used for confirming the 2-DE and mass spectrometry results by using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and enzyme linked immunosorbent assay (ELISA). 2-DE combined with mass spectrometry revealed that calreticulin (CRT) expressed normally within platelets from healthy controls, while it reduced within platelets from patients with chronic ITP. qPCR and ELISA confirmed that CRT was decreased at both RNA transcription and protein expression levels within platelets from chronic ITP patients compared with healthy controls. Decreased transcription and expression of CRT within platelets may play an important role in the pathogenesis of chronic ITP, which is worthy of further study.

摘要

观察成年慢性免疫性血小板减少性紫癜(ITP)患者与健康成年人之间蛋白质的差异。30例慢性ITP患者和30名健康对照者纳入本研究。分别从10例慢性ITP患者和10名健康对照者的外周静脉血中提取血小板总蛋白,进行二维电泳(2-DE)以找出慢性ITP患者与健康对照者之间的差异蛋白点,然后通过质谱鉴定差异蛋白点。随后,分别从另外20例慢性ITP患者和20名健康对照者中分离血小板RNA和蛋白质,并通过逆转录定量聚合酶链反应(RT-qPCR)和酶联免疫吸附测定(ELISA)用于确认2-DE和质谱结果。二维电泳结合质谱显示,钙网蛋白(CRT)在健康对照者的血小板内正常表达,而在慢性ITP患者的血小板内表达降低。qPCR和ELISA证实,与健康对照者相比,慢性ITP患者血小板内CRT在RNA转录和蛋白质表达水平均降低。血小板内CRT转录和表达降低可能在慢性ITP发病机制中起重要作用,值得进一步研究。

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