血小板裂解液与血小板凝胶生长因子谱的表达差异研究*

doi:10.3969/j.issn.1671-2587.2023.04.004

临床输血与检验 ›› 2023, Vol. 25 ›› Issue (4): 450-455.DOI: 10.3969/j.issn.1671-2587.2023.04.004

血小板裂解液与血小板凝胶生长因子谱的表达差异研究^*

肖军, 雷慧芬, 李翠莹

空军军医大学空军特色医学中心输血科,北京 100142

收稿日期:2023-07-28 出版日期:2023-08-20 发布日期:2023-09-18
通讯作者: 李翠莹,主要从事临床输血与免疫血液学研究,（E-mail）licuiying2013@qq.com。
作者简介:肖军,主要从事临床输血与免疫血液学研究,（E-mail）ammsxj@fmmu.edu.cn。
基金资助:
*本课题受中国输血协会威高科研基金（No.CSBT-WG-2021-03）资助

The Differential of Growth Factor Expression Profile in Platelet-rich Plasma Lysate and Platelet Gel

XIAO Jun, LEI Huifeng, LI Cuiying

Department of Blood Transfusion, Air force Medical Center, Beijing 100142

Received:2023-07-28 Online:2023-08-20 Published:2023-09-18

摘要/Abstract

摘要： 目的研究富血小板血浆（PRP）在不同制备条件下生长因子表达谱的变化,为临床治疗选择不同类型PRP提供实验依据。方法使用血液成分分离机单采7份PRP（30 mL/份）,同一份PRP各留取10 mL并随机分成贫血小板血浆（PPP）对照组、PRP凝胶上清组和PRP裂解液组,PRP凝胶上清组采用牛凝血酶和葡萄糖酸钙制成预混剂,预混剂与PRP按照1∶9的比例添加以获取PRP上清液;PRP裂解液组采用在-20℃和37℃反复冻存融化5次,获取血小板裂解液。应用Quantibody^®生长因子蛋白芯片检测获取的样品中生长因子谱的变化,并采用ELISA对差异生长因子进行验证。结果与PPP对照组比较,在PRP凝胶上清组中共有5个生长因子表达升高,分别为脑源性神经营养因子（BDNF,14.64倍）,血小板衍生生长因子（PDGF-AA,4.53倍）,表皮细胞生长因子（EGF,4.52倍）,血管内皮生长因子（VEGF,3.45倍）和肝细胞生长因子（HGF,2.16倍）,GO和KEGG分析表明这些因子主要富集于细胞迁移和Ras及PI3K-Akt信号通路激活,参与组织修复;而在PRP裂解液组中升高的生长因子分别为BDNF（20.74倍）,EGF（11.12倍）,转化生长因子-β1（TGF-β1,5.76倍）,减少的生长因子是胰岛素样生长因子结合蛋白-6（IGFBP-6,1.46倍）,这些因子主要富集于MAPK信号通路和FoxO信号通路发挥抗炎作用。结论 PRP凝胶上清和PRP裂解液均可释放多种生长因子;PRP凝胶上清产生的生长因子主要参与创面修复和伤口愈合,而PRP裂解液产生的生长因子主要参与组织修复和抗炎,因此针对不同的临床治疗目的,应选择不同方式制备的PRP制品,以达到最佳的治疗效果。

关键词: 富血小板血浆, 生长因子, 血小板凝胶, 血小板裂解液, 蛋白芯片

Abstract: Objective To investigate the changes in the expression profile of growth factors in platelet-rich plasma (PRP) under different preparation conditions, providing experimental evidence for selecting different types of PRP for different clinical treatment purposes. Methods Seven portions of PRP (30 mL each) were obtained using a blood component separator. Ten milliliters of the same PRP were randomly divided into platelet-poor plasma (PPP) control group, PRP gel supernatant group, and PRP frozen lysate group. The PRP gel group utilized bovine thrombin and 10% calcium gluconate to create a premix solution, which was then added to PRP in a 1∶9 ratio to obtain PRP supernatant. The PRP frozen lysate group went through five cycles of freezing and thawing at temperatures ranging from -20℃ to 37℃ to obtain platelet lysate. Changes in the growth factor profile in the samples were detected using Quantibody® growth factor protein chips, and differential growth factors were validated using ELISA. Results Compared to PPP, five growth factors showed increased expression in the PRP supernatant group: brain-derived neurotrophic factor (BDNF, 14.64 times), platelet-derived growth factor-AA (PDGF-AA, 4.53 times), epidermal growth factor (EGF, 4.52 times), vascular endothelial growth factor (VEGF, 3.45 times), and hepatocyte growth factor (HGF, 2.16 times). GO and KEGG analyses indicated that these factors were mainly enriched in cell migration, Ras, and PI3K-Akt signaling pathways, participating in tissue repair. In the PRP lysate group, the increased growth factors were BDNF (20.74 times), EGF (11.12 times), transforming growth factor-β1 (TGF-β1, 5.76 times), while the decreased growth factor was insulin-like growth factor-binding protein-6 (IGFBP-6, 1.46 times). These factors were primarily enriched in the MAPK signaling pathway and FoxO signaling pathway, playing anti-inflammatory roles. Conclusion Both PRP supernatant and PRP lysate can release a variety of growth factors. Growth factors produced in PRP supernatant mainly participate in wound repair and healing, while those in PRP lysate mainly contribute to tissue repair and anti-inflammatory effects. Therefore, for different clinical treatment purposes, PRP products prepared in different ways should be selected to achieve the best therapeutic outcome.

Key words: Platelet rich plasma, Growth factors, Platelet gel, Platelet lysate, Protein chip

中图分类号:

R331.1⁺43

肖军, 雷慧芬, 李翠莹. 血小板裂解液与血小板凝胶生长因子谱的表达差异研究^*[J]. 临床输血与检验, 2023, 25(4): 450-455.

XIAO Jun, LEI Huifeng, LI Cuiying. The Differential of Growth Factor Expression Profile in Platelet-rich Plasma Lysate and Platelet Gel[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2023, 25(4): 450-455.

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血小板裂解液与血小板凝胶生长因子谱的表达差异研究^*

The Differential of Growth Factor Expression Profile in Platelet-rich Plasma Lysate and Platelet Gel

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