title: Establishment of stable reporter expression for in vivo imaging of nuclear factor-κB activation in mouse liver.
abstract: The nuclear factor-κB (NF-κB) signaling pathway plays a critical role in a multitude of cellular processes. Activation of the NF-κB transcription factor family is essential for the initiation of inflammation, immunity, cell proliferation and apoptosis through a list of responsive genes. In hepatic tissue, activation of the NF-κB pathway has been implicated in a number of pathological conditions. Here we described a mouse model for noninvasive quantification of NF-κB activation in the hepatic tissues. Mice were subjected to hydrodynamic delivery with a mixture of pattB-NF-κB-Fluc reporter and φC31o integrase vector. Hepatic expression of φC31o integrase mediated chromosomal integration of the pattB-NF-κB-Fluc reporter, resulting in stable luciferase expression at 300 days post transfection. We applied noninvasive imaging and were able to detect NF-κB activation under acute liver injury and hepatitis conditions. During hepatectomy-induced liver regeneration, NF-κB activation was detected locally in the tissues at the surgery site. Treatment with Sorafenib suppressed NF-κB activation, accompanied with perturbation of liver regeneration. In conclusion, we established a method for stable transfection of the hepatic tissues and applied the transfected mice to longitudinal monitoring of NF-κB activity under pathological conditions. Further exploration of this methodology for establishment of other disease models and for evaluation of novel pharmaceuticals is likely to be fruitful.
authors: 1. Beijing Institute of Transfusion Medicine, 27(9) TaiPing Road, Beijing, China;
authors: Shaoduo
authors: S
authors: Yan
authors: Qiuxia
authors: Q
authors: Fu
authors: Yong
authors: Y
authors: Zhou
authors: Ning
authors: N
authors: Zhang
authors: Qianqian
authors: Q
authors: Zhou
authors: Xiaoying
authors: X
authors: Wang
authors: Zhennan
authors: Z
authors: Yuan
authors: Xiaohui
authors: X
authors: Wang
authors: Juan
authors: J
authors: Du
authors: Jingang
authors: J
authors: Zhang
authors: Linsheng
authors: L
authors: Zhan
mesh_terms: D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods*; D017930:Genes, Reporter; D005822:Genetic Vectors; D042822:Genomic Instability; D008099:Liver / Q000473:pathology*; D008107:Liver Diseases / Q000473:pathology*; D008115:Liver Regeneration / Q000502:physiology; D049409:Luciferases, Firefly / Q000032:analysis / Q000235:genetics; D051379:Mice; D016328:NF-kappa B / Q000032:analysis*
chemical_list: D016328:NF-kappa B; D049409:Luciferases, Firefly
keywords: Nuclear factor-κB; Sorafenib; animal model.; noninvasive molecular imaging; φC31o integrase