中国输血杂志2025，Vol.38Issue(3)：343-351,374,10.DOI:10.13303/j.cjbt.issn.1004-549x.2025.03.007
不同激活条件下PRR源性外泌体特征及其对HMEC-1和BJ增殖能力差异的比较研究
Characteristics of PRR-derived exosomes and the proliferation abilities of HMEC-1 and BJ under different activa-tion conditions:a comparative study
摘要
Abstract
Objective To compare the characteristics of platelet-rich plasma derived exosomes(PRP-Exos)under dif-ferent activation conditions and their differential effects on the proliferation capacit of human microvascular endothelial cells(HMEC-1)and human skin fibroblasts(BJ).Methods Ten healthy volunteers were recruited,and 10 mL of venous blood anticoagulated with EDTA-K2 was collected.PRP-Exos were prepared and extracted by the secondary centrifugation method.Transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA),and Western blotting(WB)de-tection techniques were used to compare the morphological characteristics,particle size distribution,concentration,and the expression of surface marker proteins of PRP-Exos.Through in vitro cell culture,the differences in morphological changes and proliferation abilities of HMEC-1 and BJ cells induced by PRP-Exos in different activator groups were compared.Re-sults 1)TEM observation showed that the morphologies of PRP-Exos in different activator groups were different.2)NTA analysis showed that compared with the particle size of(109.60±2.71)nm in the normal saline group,the particle sizes of the thrombin group[(104.93±1.55)nm]and the mixed agent group[(103.83±1.58)nm]were relatively smaller,while the particle size of the calcium gluconate group[(113.57±4.70)nm]was larger and its particle size distribution range was wider.There were statistically significant differences among different groups(F=7.019,P<0.05).The concen-tration of exosomes obtained in the group with the mixture of calcium gluconate and thrombin was the highest[(4.87±0.63)×106 particles/mL,P<0.001].Both the thrombin group[(2.75±0.13)×106 particles/mL,P<0.01]and the calcium gluconate group[(3.82±0.06)×106 particles/mL,P<0.001]obtained higher concentrations of exosomes com-pared with the normal saline group.The concentration in the calcium gluconate group was slightly higher than that in the thrombin group,and there was a significant difference between the two groups(P<0.05).3)WB analysis showed that CD41,CD81,and TSG101 were expressed on the surface of PRP-Exos,and the protein signal intensities of CD41 and TSG101 in the group with the mixture of thrombin and calcium gluconate were the strongest(P<0.001).4)The results of in vitro cell culture showed that PRP-Exos in the group with the mixture of thrombin and calcium gluconate could significant-ly promote the proliferation of HMEC-1 and BJ cells(P<0.05).Conclusion The PRP treated with the thrombin and calci-um gluconate mixture group yielded the highest concentration of exosomes;under in vitro culture conditions,PRP-Exos from this group significantly promoted the proliferation of HMEC-1 and BJ cells.关键词
PRP/外泌体/凝血酶/葡萄糖酸钙/人微血管内皮细胞/人皮肤成纤维细胞/细胞增殖Key words
PRP/exosomes/thrombin/calcium gluconate/human microvascular endothelial cells/human skin fibro-blasts/cell proliferation分类
医药卫生引用本文复制引用
高立兰,吕孟兴,刘建香,胡美坤,金晓红,屈柯暄..不同激活条件下PRR源性外泌体特征及其对HMEC-1和BJ增殖能力差异的比较研究[J].中国输血杂志,2025,38(3):343-351,374,10.基金项目
儿童富血小板血浆制备的影响因素及质量控制的研究(2023-03-236-K01) （2023-03-236-K01）
