不同来源人血清白蛋白对血管内皮细胞生物学效应的影响研究

Biological Effects of Human Serum Albumin from Different Sources on Vascular Endothelial Cells

目的研究不同来源人血清白蛋白(human serum albumin,HSA)对血管内皮细胞生物学效应的影响。方法对体外无血清培养的HUVEC细胞添加不同来源HSA处理,利用CCK-8细胞活性检测试剂盒、Annexin V-FITC细胞凋亡检测试剂盒和细胞周期与细胞凋亡检测试剂盒,检测不同来源HSA处理后各组细胞增殖活性、细胞凋亡情况和细胞周期分布,并分析比较各组间差异。通过细胞划痕实验和管形成实验分析比较不同来源HSA处理对HUVEC细胞迁移和管形成的影响。结果酵母来源rHSA1对HUVEC细胞增殖活性无影响(P=0.49,q=1.601),其他HSA处理组均显示出显著促进HUVEC细胞增殖的活性,且不同来源HSA处理组间的HUVEC增殖活性存在显著差异(P<0.001,F=10.84)。所有HSA处理组均显示出抑制HUVEC细胞凋亡的作用(P<0.001),不同处理组间活细胞(P=0.07,F=2.415)、凋亡细胞(P=0.2,F=1.624)和坏死细胞(P=0.28,F=1.376)比例均无统计学差异。对比无血清对照组,在HSA处理组中除pHSA2和pHSA6处理组G0/G1期细胞比例显著降低(P<0.001)、S期无明显变化、G2/M期细胞比例明显升高(P<0.01)外,其他HSA处理组各细胞周期比例均无明显改变。在划痕实验中,对比无血清对照组,HSA处理组中pHSA(1)和pHSA(2)处理组经过12 h(P<0.001)、24 h(P<0.001)、36 h(P<0.001,P=0.002)后划痕愈合程度更高,差异具有统计学意义,而rHSA(1)和rHSA(2)处理组较对照组无明显差异。在管形成实验中,对比无血清对照组,HSA处理组中pHSA(1)和pHSA(2)处理组的节点数量(P=0.001、P=0.005)、管状分支(P<0.001)及网状结构(P<0.001)均显著增多,而rHSA(1)和rHSA(2)处理组较对照组无统计学差异。结论不同来源HSA均能够显著抑制无血清条件下血管内皮细胞的凋亡,且对血管内皮细胞的增殖活性、细胞周期、细胞迁移和管形成的影响存在一定差异。

Objective To evaluate the differential biological effects of human serum albumin(HSA)derived from different sources on vascular endothelial cells.Methods Human umbilical vein endothelial cells(HUVEC)were serum-starved and then exposed to HSA from varied sources.Cellular responses were assessed using the CCK-8 assay for proliferation,Annexin V-FITC kit for apoptosis,and flow cytometry for cell cycle analysis.The proliferative,apoptotic and cell cycle effects of HSA from different sources were statistically compared.Additionally,the impact of HSA on HUVEC cell migration and tube formation was assessed via scratch assays and tube formation experiments.Results Yeast-derived rHSA1 did not affect HUVEC proliferation(P=0.49,q=1.601),while other HSA sources significantly promoted it(P<0.001,F=10.84).All HSA treatment groups exhibited an inhibitory effect on HUVEC apoptosis(P<0.001),with no statistically significant differences in the proportions of live cells(P=0.07,F=2.415),apoptotic cells(P=0.2,F=1.624),and dead cells(P=0.28,F=1.376)across treatment groups.Compared to the serum-free control group,except for the pHSA2 and pHSA6 treatment groups,which showed a significant decrease in G0/G1 phase cell proportion(P<0.001)and a significant increase in G2/M phase cell proportion(P<0.01),the proportions of cells in other cell cycle phases did not show significant changes in the HSA treatment groups.In the scratch assay,in contrast to the serum-free control group,the pHSA(1)and pHSA(2)groups exhibited a higher degree of wound healing at 12 h(P<0.001),24 h(P<0.001),and 36 h(P<0.001,P=0.002);however,no significant differences were observed in the rHSA(1)and rHSA(2)groups compared to the control group.Likewise,in the tube formation assay,the pHSA(1)and pHSA(2)groups significantly enhanced node formation(P=0.001,P=0.005),tubular branching(P<0.001),and mesh structure development(P<0.001),whereas the rHSA(1)and rHSA(2)groups showed no such enhancements.Conclusion HSA from different sources significantly inhibits endothelial cell apoptosis under serum-free conditions,but exerts varying effects on proliferation,cell cycle regulation,cell migration,and tube formation of vascular endothelial cells.