CpG-N ODN对CpG-S ODN诱导hPBMC释放TNF-α抑制作用.docVIP

CpG?N ODN对CpG?S ODN诱导hPBMC释放TNF?α抑制作用 　 作者：蒋为薇 王良喜 丁国富 余双江 毕业论文 【摘要】; 目的 明确抑制性CpG ODN 208(CpG?N ODN)对刺激性CpG ODN 1826(CpG?S ODN)诱导单核/巨噬细胞分泌TNF?α的影响，为通过拮抗细菌DNA达到防治脓毒症的目的提供实验依据。方法 选用本室前期筛选出的对CpG?S ODN活化RAW 264.7细胞有抑制作用的CpG?N ODN，观察其对CpG?S ODN诱导hPBMC释放TNF?α的影响；应用流式细胞术观察CpG?N ODN对CpG?S ODN与hPBMC表面结合和内化的影响。结果 浓度比为1∶1的CpG?N ODN对刺激性CpG?S ODN诱导hPBMC分泌TNF?α具有抑制作用，并呈一定时效关系；CpG?N ODN对CpG?S ODN在hPBMC细胞表面的结合和内化具有抑制作用。结论 CpG?N ODN对CpG?S ODN活化hPBMC具有抑制作用，这个作用可能与其影响hPBMC的表面结合和内化CpG?S ODN有关。 毕业论文 【关键词】; 脓毒症 抑制性CpG ODN 刺激性CpG ODN TNF?α 表面结合 内化 毕业论文 ; ABSTRACT; Objective; To study the influence of CpG ODN 208 (CpG?N ODN) on TNF?α release from hPBMC induced by CpG?S ODN 1826 (CpG?S ODN).; Methods; CpG?N ODN, selected by the previous work in our lab, can inhibit TNF?α release from RAW264.7 induced by CpG?S ODN. The influence of CpG?N ODN on TNF?α release from hPBMC induced by CpG?S ODN was measured by ELISA. The effects of CpG?N ODN on the surface binding and internalization of CpG?S ODN were tested by flow cytometry.; Results CpG?N ODN could inhibit TNF?α release from hPBMC induced by CpG?S ODN at the concentration ratio 1∶1 in a time?dependent manner. CpG?N ODN could inhibit the surface binding and internalization of CpG?S ODN. Conclusion; CpG?N ODN could inhibit TNF?α release from hPBMC induced by CpG?S ODN, which partly attributed to inhibition of CpG?N ODN on the surface binding and internalization of CpG?S ODN. 毕业论文 ; KEY WORDS; Sepsis CpG?N ODN CpG?S ODN TNF?α Surface binding Internalization 毕业论文 ; 脓毒症(sepsis)是一种严重创伤和感染的引起全身炎症反应综合征(systemic inflammatory response syndrome，SIRS)，由其发展来的多器官功能不全综合征(multiple organ dysfunction syndrome，MODS)病死率高达30%～50%，但目前尚无有效的防治措施［1］。细菌基因组DNA(细菌DNA)是SIRS的主要启动因子之一。未甲基化CpG的寡核苷酸片段(CpG oligonucleotides，CpG ODN)是细菌DNA的免疫刺激作用的最小作用单位，广泛存在于细菌DNA中［2］。细菌DNA之所以被哺乳动物视作异己成分而启动防御反应，是因为其有着与不如DNA明显不同的结构特征：细菌DNA中的CpG二核苷酸出现的频率约为1/16，而哺乳动物DNA中的CpG二核苷酸出现的频率仅为1/64，并且80%发生甲基化［3］。研究者们发现只有含有未甲基化CpG为核心的特定六核苷酸序列的CpGODN才具有刺激性， 这些特定的六核苷酸被称为刺激性CpG基序(stimul