【摘要】：目的 構建人源性髓樣細胞表達的激活受體-1(TREM-1)胞外段與免疫球蛋白G1 Fc段(IgG1 Fc)融合基因并表達融合蛋白,評價TREM-1-IgG1 Fc融合蛋白對內毒素休克的保護作用。方法 設計引物分別從人外周血淋巴細胞及人單核白血病細胞系(U937)擴增IgG1 Fc段及TREM-1胞外段序列,通過酶切位點將兩段基因連入載體pcDNA3.1/myc-his(-)B。通過轉染小鼠成纖維細胞系(NIH/3T3)獲得穩(wěn)定表達融合蛋白的細胞株。BALB/C小鼠經腹腔注射1μg LPS和20mg D-Gal建立內毒素休克模型,并分別在給藥前1h,給藥后1h、2h、4h、6h時給予純化的TREM-1-Ig融合蛋白,觀察小鼠72h存活率。給予LPS和D—Gal后1h、2h、4h、6h、8h取小鼠眼眶靜脈血,檢測血清中TNF-α、IL-1β水平。結果 在LPS給藥前后短時間內給予TREM-1-Ig融合蛋白,均可顯著降低內毒素休克小鼠的死亡率,并可明顯降低外周血TNF-α、IL-1β濃度(P0.01)。結論TREM-1-Ig融合蛋白可緩解內毒素休克炎癥反應進程,對內毒素休克小鼠有顯著的保護作用。
[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- 偽 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- 偽 -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.
【學位授予單位】：第二軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R392

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本文編號：2210394