人源性TREM-1-Ig融合蛋白对内毒素休克小鼠的保护作用
发布时间：2018-08-28 19:33
【摘要】：目的 构建人源性髓样细胞表达的激活受体-1(TREM-1)胞外段与免疫球蛋白G1 Fc段(IgG1 Fc)融合基因并表达融合蛋白,评价TREM-1-IgG1 Fc融合蛋白对内毒素休克的保护作用。方法 设计引物分别从人外周血淋巴细胞及人单核白血病细胞系(U937)扩增IgG1 Fc段及TREM-1胞外段序列,通过酶切位点将两段基因连入载体pcDNA3.1/myc-his(-)B。通过转染小鼠成纤维细胞系(NIH/3T3)获得稳定表达融合蛋白的细胞株。BALB/C小鼠经腹腔注射1μg LPS和20mg D-Gal建立内毒素休克模型,并分别在给药前1h,给药后1h、2h、4h、6h时给予纯化的TREM-1-Ig融合蛋白,观察小鼠72h存活率。给予LPS和D—Gal后1h、2h、4h、6h、8h取小鼠眼眶静脉血,检测血清中TNF-α、IL-1β水平。结果 在LPS给药前后短时间内给予TREM-1-Ig融合蛋白,均可显著降低内毒素休克小鼠的死亡率,并可明显降低外周血TNF-α、IL-1β浓度(P0.01)。结论TREM-1-Ig融合蛋白可缓解内毒素休克炎症反应进程,对内毒素休克小鼠有显著的保护作用。
[Abstract]:Objective to construct the fusion gene of extracellular activated receptor 1 (TREM-1) and immunoglobulin G1 Fc (IgG1 Fc) expressed by human myeloid cells and express the fusion protein and evaluate the protective effect of TREM-1-IgG1 Fc fusion protein on endotoxic shock. Methods primers were designed to amplify the IgG1 Fc fragment and the extracellular segment of TREM-1 from human peripheral blood lymphocytes and human monocytic leukemia cell line (U937), respectively. The two fragments were inserted into the vector pcDNA3.1/myc-his (-) B by restriction endonuclease site. The model of endotoxic shock was established by intraperitoneal injection of 1 渭 g LPS and 20mg D-Gal in BALB / C mice transfected with mouse fibroblast cell line (NIH/3T3), and the purified TREM-1-Ig fusion protein was given at 1 hour before administration and 1 h after administration for 4 h or 6 h, respectively. 72 h survival rate of mice was observed. The serum levels of TNF- 伪 and IL-1 尾 were measured after administration of LPS and D-Gal at 1h, 2h, 4h, 6h and 8h, respectively. Results the TREM-1-Ig fusion protein before and after administration of LPS could significantly reduce the mortality of mice with endotoxic shock and the concentration of TNF- 伪 -IL-1 尾 in peripheral blood (P0.01). Conclusion TREM-1-Ig fusion protein can attenuate the inflammatory process of endotoxic shock and has a significant protective effect on endotoxic shock mice.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392
本文编号：2210394
[Abstract]:Objective to construct the fusion gene of extracellular activated receptor 1 (TREM-1) and immunoglobulin G1 Fc (IgG1 Fc) expressed by human myeloid cells and express the fusion protein and evaluate the protective effect of TREM-1-IgG1 Fc fusion protein on endotoxic shock. Methods primers were designed to amplify the IgG1 Fc fragment and the extracellular segment of TREM-1 from human peripheral blood lymphocytes and human monocytic leukemia cell line (U937), respectively. The two fragments were inserted into the vector pcDNA3.1/myc-his (-) B by restriction endonuclease site. The model of endotoxic shock was established by intraperitoneal injection of 1 渭 g LPS and 20mg D-Gal in BALB / C mice transfected with mouse fibroblast cell line (NIH/3T3), and the purified TREM-1-Ig fusion protein was given at 1 hour before administration and 1 h after administration for 4 h or 6 h, respectively. 72 h survival rate of mice was observed. The serum levels of TNF- 伪 and IL-1 尾 were measured after administration of LPS and D-Gal at 1h, 2h, 4h, 6h and 8h, respectively. Results the TREM-1-Ig fusion protein before and after administration of LPS could significantly reduce the mortality of mice with endotoxic shock and the concentration of TNF- 伪 -IL-1 尾 in peripheral blood (P0.01). Conclusion TREM-1-Ig fusion protein can attenuate the inflammatory process of endotoxic shock and has a significant protective effect on endotoxic shock mice.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392
【参考文献】
相关期刊论文 前2条
1 李勇,韩本立,王会信,龙建银,李昆;基因转移人TGF-β_1在NIH/3T3细胞中表达鉴定[J];第三军医大学学报;2000年01期
2 余鹰,周鸣,曹利,唐珂,李伟芳,李桂源;pcDNA3.1表达载体转染对细胞生长的影响[J];生物技术;2001年01期
，本文编号：2210394
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