高迁移率族蛋白1对脐静脉内皮细胞迁移及相关蛋白表达的影响

Effects of High Mobility Group Box 1 on Migration and Expression of Related Proteins in Umbilical Vein Endothelial Cells

【目的】组织工程中微循环的建立在生物材料植入中发挥重要作用,其中血管内皮细胞迁移是快速血管生成的主要影响因素。组织和器官损伤后常引发炎症反应,高迁移率族蛋白1(HMGB1)是炎症反应的起始因子,但对内皮细胞迁移的影响尚不清楚。【方法】体外提取和培养人脐静脉内皮细胞(HUVECs),并进行细胞表型鉴定;构建培养液中含有HMGB1的浓度梯度,通过CCK-8、划痕实验和Transwell小室实验检测HUVECs的增殖能力和迁移能力,采用RT-qPCR和Western Blot检测细胞内细胞增殖因子和迁移因子基因和蛋白的表达情况。【结果】不同浓度HMGB1对HUVECs的增殖能力无显著影响,但对细胞迁移能力有浓度依赖性,随着HMGB1浓度增加,细胞的迁移能力增强。FGF-2表达随HMGB1浓度增加表达上调;高浓度HMGB1处理HUVECs后,VEGFA表达明显下降。【结论】HMGB1可通过FGF-2促进HUVECs迁移,为今后HMGB1对内皮细胞迁移能力的影响研究提供依据。

【Purposes】The establishment of microcirculation in tissue engineering plays an important role in biomaterials implantation,in which the migration of vascular endothelial cells is the main factor affecting rapid angiogenesis.Tissue and organ injury often leads to inflammation,and high mobility group box 1(HMGB1)is the initiating factor of inflammation,while its effect on the migration of endothelial cells remains unclear.【Methods】Human umbilical vein endotheli-al cells(HUVECs)were extracted and cultured in vitro,and their phenotypes were identified.The concentration gradient of HMGB1 in culture medium was constructed,and the proliferation and migration of HUVECs were detected by CCK-8,scratch test,and Transwell chamber test.The expression of genes and proteins of cell proliferation factor and migration factor in HUVECs was detected by RT-qPCR and Western Blot.【Findings】Different concentrations of HMGB1 had no significant effect on proliferation of HUVECs,but was concentration-dependent on cell migra-tion.With the increase of HMGB1 concentration,the migration ability of cells was enhanced.The expression of FGF-2 was up-regulated with the increase of HMGB1 concentration.The ex-pression of VEGFA decreased significantly after HUVECs was treated with high HMGB1 con-centration.【Conclusions】HMGB1 can promote the migration of HUVECs through FGF-2,which provides a basis for the study of the effect of HMGB1 on the migration ability of endotheli-al cells in the future.