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

作　　者：肖军 雷慧芬 李翠莹 XIAO Jun;LEI Huifeng;LI Cuiying(Department of Blood Transfusion,Air force Medical Center,Beijing 100142)

机构地区：[1]空军军医大学空军特色医学中心输血科,北京100142

出　　处：《临床输血与检验》2023年第4期450-455,共6页Journal of Clinical Transfusion and Laboratory Medicine

基　　金：中国输血协会威高科研基金(No.CSBT-WG-2021-03)资助。

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。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

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 

分 类 号：R331.143[医药卫生—人体生理学]