反义TNF_(-α)寡脱氧核苷酸降低感染性休克小鼠死亡率

Antisense TNF-α oligodeoxynucleotide decreases death rate in septic shock mice

目的：观察反义肿瘤坏死因子－α（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ－ａｌｐｈａ，ＴＮＦ－α）寡脱氧核苷酸对ＬＰＳ诱导的感染性休克小鼠的影响，为采用反义技术降低感染性休克死亡率的早期临床试验奠定基础。方法：通过腹腔注射ＬＰＳ建立感染性休克小鼠模型。采用腹腔注射方法，通过ＤＯＴＡＰ转染导入系统，使反义ＴＮＦ－α寡脱氧核苷酸（ｏｌｉｇｏｄｅｏｘｙｎｕｃｌｅｏｔｉｄｅ，ＯＤＮ）作用于小鼠单核吞噬细胞系统；采用ＲＴ－ＰＣＲ，ＥＬＩＳＡ检测ＴＮＦ－α的ｍＲＮＡ和蛋白质的表达水平。结果：反义ＴＮＦ－αＯＤＮ降低了ＬＰＳ诱导的感染性休克小鼠的ＴＮＦ－α的释放，降低了感染性休克小鼠的死亡率，而正义ＴＮＦ－αＯＤＮ和错配ＴＮＦ－αＯＤＮ与对照组相比，对ＴＮＦ－α的释放无影响，未能降低感染性休克小鼠的死亡率。结论：反义ＴＮＦ－αＯＤＮ以序列特异性方式抑制了ＬＰＳ诱导的感染性休克小鼠的ＴＮＦ－α的释放，降低了感染性休克小鼠的死亡率。

AIM:To evaluate the effect of antisense phosphrothioate tumor necrosis factor-alpha(TNF-α) oligodeoxynucleotide(ODN) on death rate in septic mice induced by LPS.METHODS: Septic shock mice model was induced by intraperitoneal injection of LPS.Mouse antisense TNF-α ODN was applied to the reticular-endothelial system in septic shock BALB/c mice introduced by DOTAP delivery system . RT-PCR and ELISA were used to detect TNF-α mRNA and protein.RESULTS: The antisense TNF-α ODN can suppress releasing of TNF-α and reduce death rate in septic shock mice induced by LPS, and neither sense nor mismatched TNF-α ODN can. CONCLUSION: Antisense TNF-α ODN significantly suppressed production of TNF-α in septic shock.The investigation provides the basis for the early clinical trials assessing the therapeutic potential of antisense TNF-α approach for decreasing death rate in septic shock.