人HMGB1诱导单核细胞分泌TNF-α的规律

TNF-α secretion in human THP1 cells induced by human HMGB1

目的 :探讨人高迁移率族蛋白 1 (HMGB1 )诱导人单核细胞TNF α的分泌规律和HMGB1在脓毒症中的作用 .方法 :HMGB1的剂量效应组分别用含有 0 .1 ,1 .0 ,1 0 ,2 0和5 0mg·L-1 HMGB1的培养基孵育与人单核巨噬细胞系THP1 ,共培养 6h ,用双抗体夹心ELISA法测其上清中的TNF α含量 ,采用流式细胞技术观察细胞的凋亡 .HMGB1的时间效应组以含有 2 0mg·L-1 HMGB1的培养基、对照组以不含HMGB1的培养基、另外以含有 2 0mg·L-1 HMGB1和10 0mg·L-1 脂肪多糖 (lipopolysaccharide ,LPS)的混合组的培养基分别培养THP1细胞 0 ,2 ,4 ,6 ,8,1 0 ,1 2和 2 4h ,同上留取离心后的培养液上清 ,用双抗体夹心ELISA法测其上清中的TNF α含量 .结果 :将HMGB1加入到单核细胞培养液中能明显刺激其TNF α的分泌 .在 0 .1～ 5 0mg·L-1 范围内 ,HMGB1诱导THP1的TNF α分泌增加 ,呈显著剂量依赖性关系 ,5 0mg·L-1 HMGB1能明显诱导THP1细胞的凋亡 ;HMGB1组和HMGB1与LPS的混合组THP1的TNF α分泌在 0～ 2 4h间呈双相性规律 .结论 :人HMGB1能诱导人单核细胞的TNF α分泌与凋亡 ,初步证实HMGB1在脓毒症的发生。

AIM: To explore the secretion pattern of TNF α in human THP1 cells induced by human HMGB1 and the function of human HMGB1 in sepsis. METHODS: One group of THP1 cells were cultured with human HMGB1 in 0.1 , 1.0, 10, 20 and 50 mg·L -1 for 6 h. THP1 cells of the control group were cultured with 20 mg·L -1 HMGB1 alone or 20 mg·L -1 HMGB1 mixed with 100 mg·L -1 lipopolysaccharide (LPS) for 0, 2, 4, 6, 8, 10, 12 and 24 h respectively. The quantities of TNF α secreted by THP1 cells were measured by sandwich ELISA and the apoptosis of THP1 cells was observed by flow cytometry. RESULTS: HMGB1 markedly induced the release of TNF α from THP1 cells in a dose dependent manner from 0.1 mg·L -1 to 50 mg·L -1 HMGB1. There were two peaks of TNF α after THP1 cells were induced by HMGB1 alone or mixed with LPS in the period of 0～24 h. 50 mg·L -1 HMGB1 markedly effected apoptosis of THP1 cells. CONCLUSION: Human HMGB1 can markedly induce the release of TNF α from THP1 cells and effect apoptosis of THP1 cells, indicating that human HMGB1 may play an important role in the occurrence and development of sepsis.