重组人肿瘤坏死因子对脐血单个核细胞明胶酶B活性及体外迁移能力的影响

Effect of Recombinant Human Tumor Necrosis Factor Alpha on Activity of Gelatinase B and Migration of Human Umbilical Cord Blood Mononuclear Cells In Vitro

本研究探讨脐血单个核细胞(mononuclear cells,MNC)中基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的活性表达,重组人肿瘤坏死因子-α(recombinant human tumor necrosis factor alpha,rh-TNF-α)对脐血MNC MMP-9活性和细胞体外迁移能力中的作用及二者之间的关系。采用明胶酶谱技术检测脐血MNC中MMP-9的活性。通过微孔细胞迁移实验检测脐血MNC的体外迁移能力,并以不同浓度的rh-TNF-α对脐血MNC进行干预,检测rh-TNF-α对MMP-9活性和细胞体外迁移的作用。结果表明:脐血MNC经无血清培养24和48小时后,均可检测出MMP-9活性,细胞相对迁移率分别为(1.371±0.011)%和(1.384±0.014)%,两者间无统计学差异。大剂量TNF-α(500U/ml)作用可致细胞死亡;小剂量TNF-α(<200U/ml)可以上调脐血MNC MMP-9表达,且随着TNF-α剂量增加MMP-9表达增强。小剂量TNF-α(<200U/ml)可提高脐血单个核细胞的体外迁移率,且与剂量呈正相关。结论:小剂量TNF-α可上调脐血MNC MMP-9活性,提高脐血MNC的体外迁移能力,此作用可能加速脐血移植后细胞归巢,促进脐血移植后的造血重建。

This study was aimed to investigate the acitivity of matrix metalloproteinase-9 （ MMP-9 ） in umbilical cord blood mononuclear cells, the effect of recombinant human tumor necrosis factor alpha （rhTNF-α） on the activity of MMP-9 and cell migration capability of umbilical cord blood mononuclear cells, as well as the relationship between them. The different concentrations of TNF-α was added into the mononuclear cells. The activity of MMP-9 in umbilical cord blood mononuclear cells were analyzed by gelatinase spectrum. The cell migration capability was observed through the milliceU carbine. The results showed that after umbilical cord blood mononuclear ceils were cultured in serum-free RPMI 1640 medium for 24 and 48 hours, the activity of MMP-9 could be detected, the cell migration capability at 24 and 48 hours were （ 1.371 ± 0.011 ） % and （ 1. 384 ± 0. 014） % respectively, there was no statistical difference between them. Large dose of TNF-α （500 U/ml） led to the death of the cells; small doses of TNF-α （ 〈 200 U/ml） upregulated the acitivity of MMP-9, and as the concentration of TNF-α increased, the acitivity of MMP-9 was enhanced. Small doses of TNF-α （ 〈 200 U/ml） improved the migration capability of umbilical cord blood mononuclear cells in a dose-dependent manner. It is concluded that small doses of TNF-α can upregulate the activity, enhance the migration capability of umbilical cord blood mononuclear cells, which probably accelerates cell homing after umbilical cord blood stem cell transplant and improves the hematopoietic reconstruction after transplantation.