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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。
Objective To establish a mouse model of real-time, non-invasive and specific monitoring of the transient expression of hepatic interleukin-6 (IL-6). Methods Fluorescent luciferase (Fluc) reporter gene promoter initiated by IL-6 promoter was transfected into mouse liver by hydrodynamic high-pressure transfection. The expression of liver Fluc in inflammatory stimuli was detected by bioluminescence imaging. The correlation between luminescence intensity and hepatic IL-6 expression was also investigated. Results High-pressure hydrodynamic method successfully delivered p IL-6-Fluc reporter gene to mouse liver and approximately 20% of hepatic parenchymal cells expressed the target gene. The law can cause transient liver damage but can return to normal in 5 ~ 7 days. p IL-6-Fluc was activated by lipopolysaccharide (LPS), which increased by 46.80 ± 13.35 folds at the peak, much higher than that of the ELISA assay [(4.09 ± 0.96) fold]. CONCLUSION: The pIL-6-Fluc can be specifically delivered to mouse liver by hydrodynamic transfection of tail vein and the transient expression model of liver IL-6 in mice can be established by live imaging method. The model can be used to confirm the specificity Reflect IL-6 activation, and its sensitivity is higher than the classical ELISA method.