高压水动力法建立肝特异性白细胞介素-6报告基因小鼠模型

发布时间：2018-06-17 03:12

本文选题：基因转染 + 白细胞介素-　；参考：《军事医学》2017年04期

【摘要】：目的建立一种可实时、无创、特异监测肝白细胞介素-6(IL-6)瞬时表达的小鼠模型。方法通过水动力高压转染法将IL-6启动子启动的萤火虫荧光素酶(Fluc)报告基因载体定向转染至小鼠肝,采用活体生物发光成像检测炎症刺激条件下肝Fluc的表达情况,并考察发光强度与肝IL-6表达的相关关系。结果高压水动力法成功将p IL-6-Fluc报告基因递送至小鼠肝,约使20%肝实质细胞表达目的基因。该法可造成一过性肝损伤但可在5~7 d恢复正常。p IL-6-Fluc可被脂多糖(LPS)激活,峰值时可上升(46.80±13.35)倍,远高于ELISA检测结果[(4.09±0.96)倍]。结论通过尾静脉水动力高压转染法可将p IL-6-Fluc特异性地递送至小鼠肝进而建立活体成像法检测小鼠肝IL-6瞬时表达模型;经验证,该模型可特异性地反映IL-6激活情况,且其灵敏度高于经典的ELISA方法。
[Abstract]:Objective to establish a mouse model of real-time, noninvasive and specific monitoring of the transient expression of interleukin-6 (IL-6) in the liver. Methods the fluorescent luciferase (luciferase) reporter gene vector of IL-6 promoter was transfected into mouse liver by hydrodynamic high pressure transfection. The expression of Fluc was detected by in vivo bioluminescence imaging. The relationship between the intensity of luminescence and the expression of IL-6 in liver was investigated. The method could cause transient liver injury but could be activated by lipopolysaccharide (LPS) on the 7th day, and the peak value could be increased by 46.80 卤13.35 times, which was much higher than that of Elisa [4.09 卤0.96]. Conclusion pIL-6-Fluc can be specifically delivered to mouse liver by tail vein hydrodynamic high pressure transfection method, and then establish an in vivo imaging method to detect the transient expression of IL-6 in mouse liver, and it is proved that the model can reflect the activation of IL-6 specifically. The sensitivity of Elisa is higher than that of classical Elisa.
【作者单位】：泰山医学院公共卫生学院;军事医学科学院野战输血研究所;
【基金】：国家自然科学基金面上资助项目(81371806);国家自然科学基金资助项目(81401369) 军队“十三五”重点项目(BWS16J006-08)
【分类号】：R-332

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