高迁移率族蛋白B1对16HBE细胞血管内皮生长因子表达和分泌的影响*
Effects of high？？mobility group protein B1 on expression and secretion of VEGF in 16HBE cells

摘要目的：探讨高迁移率族蛋白B1(HMGB1)对人支气管上皮细胞（16HBE）血管内皮生长因子(VEGF）表达和分泌的影响及可能调控机制。方法：16HBE按以下方法分组。①HMGB1不同质量浓度组：0、100、500、2 000 μg/L HMGB1刺激16HBE 24 h。②HMGB1不同作用时间组：对照组，2 000 μg/L HMGB1分别刺激16HBE 6、12、24 h组。MTT检测上述各组干预后16HBE的细胞活力，实时荧光定量PCR检测16HBE VEGF mRNA表达水平的变化，ELISA检测细胞培养上清中VEGF水平。应用P38 MAPK通路特异性抑制剂SB？？202190观察其对HMGB1诱导VEGF表达和分泌的影响。结果：随HMGB1作用浓度的增加和作用时间的延长，VEGF的表达和分泌呈上升趋势（P均＜0.05），但细胞活力无明显改变（P均＞0.05）。HMGB1刺激增加了16HBE细胞VEGF的表达和分泌，SB？？202190几乎完全抑制了HMGB1诱导的VEGF的表达和分泌（P均＜0.05）。结论：HMGB1可能通过P38 MAPK通路介导支气管上皮细胞VEGF的表达和分泌。
AbstractAim: To investigate the effect of high？？mobility group protein B1(HMGB1) on the expression and secretion of vascular endothelial growth factor(VEGF) in human bronchial epithelial cell line 16HBE and mechanism underlying this process. Methods: 16HBE cells were allocated into different groups according to the following methods. 1)different concentrations: 16HBE cells were coincubated with different concentrations of HMGB1(0, 100, 500, 2 000 μg/L). 2)different intervention time: 16HBE cells were incubated with HMGB1 at 2 000 μg/L for 6, 12, 24 h, respectively. MTT assay was used to assess the viability of 16HBE cells. The expression of VEGF mRNA was determined by real？？time PCR and the level of VEGF in the supernatant was determined by ELISA. The effect of P38 MAPK pathway on HMGB？？induced expression and secretion of VEGF was analyzed by use of a special inhibitor,SB？？202190. Results: The expression and secretion of VEGF had an increasing tendency with the increase of concentration and treatment time of HMGB1(P<0.05), but there was no obvious changes in 16HBE viability. P38 inhibitor SB？？202190 abolished the HMGB1？？induced VEGF expression and secretion(P<0.05). Conclusion: HMGB1 increases VEGF expression and secretion via a P38 MAPK？？dependent pathway in bronchial epithelial cells