烧伤血清诱导RAW264．7细胞分泌TNF-α的规律

Dynamics of TNF-α secretion from RAW264.7 cells induced by burn serum

目的:探讨烧伤血清诱导小鼠单核巨噬细胞系RAW264．7细胞分泌TNF-a的规律及其在脓毒症中的作用。方法:烧伤血清的剂量效应组分别用含有0．5,1．0,1．5,2．0,2．5ml烧伤血清的培养基与小鼠单核巨噬细胞系RAW264．7共培养6h,用双抗体夹心ELISA法测其上清中的TNF-a含量。采用流式细胞技术观察细胞的凋亡。烧伤血清的时间效应组以含有2．5ml烧伤血清的培养基,对照组以不含烧伤血清的培养基,另外以含有2．5ml烧伤血清和100mg／L LPS的混合组的培养基分别培养RAW264．7细胞0,2,4,6,8,10,12,24h,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-a含量。结果:将烧伤血清加入到小鼠单核巨噬细胞RAW264．7培养液中能明显刺激TNF-a的分泌。在0．5ml～2．5ml范围内,烧伤血清诱导RAW264．7细胞的TNF-a分泌增加,呈显著的剂量依赖性关系,2．5ml烧伤血清能明显诱导RAW264．7细胞的凋亡;烧伤血清组和烧伤血清与LPS的混合组RAW264．7细胞的TNF-a分泌在0～24h间呈双相性规律。结论:烧伤血清能诱导小鼠单核巨噬细胞系RAW264．7的TNF-a分泌与凋亡,初步证实烧伤血清在脓毒症的发生、发展中发挥重要作用。

Objective： To explore the dynamics of the secretion of TNF - α, from RAW264. 7 cells induced by burn serum and its effect on sepsis in mice. Method： To observe the effect of the dose of the serum on the secretion of TNF - α, RAW264. 7 cells were cultivated with respectively 0. 5, 0. 1, 1.5, 2. 0 and 2. 5 ml of burned serum for 6 hours. TNF -α content was determined by ELISA method. Cell apoptosis was observed using flow cytophotometric method. To observe the time effect of bum serum, RAW264. 7 cells were stimulated respectively with 2. 5 ml bum serum alone or with 2. 5 ml bum serum mixed with 100 mg/L LPS, for0, 2, 4, 6, 8, 10, 12 and 24 hours and the content of the released TNF - α was determined using ELISA method. Result： 0. 5 to 2. 5 ml bum serum could markedly stimulate the secretion of TNF - α from RAW 264. 7 cell and the increase in secretion is significantly related to the dose of the serum. 2. 5 ml bum serum could markedly induce RAW 264. 7 cell apeptosis. The secretion stimulated by bum serum alone and bum serum mixed with LPS had two peaks in 0 to 24 hours. Conclusion： Bum serum can markedly induce apeptosis and TNF - α secretion of RAW264. 7 cells.