Hydrodynamics-based transfection of plasmid encoding receptor activator for nuclear factor kappa B-Fc protects against hepatic ischemia/reperfusion injury in mice.
Hepatic ischemia/reperfusion (I/R) injury is very important in transplant surgery. To study the mechanism of receptor activator for nuclear factor kappa B-Fc (RANK-Fc) in protection against I/R injury, 90 male BALB/c mice were randomly divided into 3 groups: a phosphate-buffered saline (PBS) (sham) group, a pLNCX2-IRES-eGFP+I/R (Negative-control) group (where IRES means internal ribosome entry site and eGFP means enhanced green fluorescent protein), and a pLNCX2-RANK-Fc-IRES-eGFP+I/R (RANK-Fc) group. All mice were injected with 2.5 mL of PBS (with or without plasmids) within 6 seconds via the tail vein. After 3 days, hepatic I/R was induced under warm conditions by partial occlusion of the left and median lobes for 90 minutes followed by various periods of reperfusion. Hepatic injury was assessed by the levels of liver aminotransferases and histopathology. Tumor necrosis factor alpha, interleukin 6, and interleukin 1beta were measured by enzyme-linked immunosorbent assay, whereas RANK-Fc, phospho-c-Jun, c-Jun N-terminal kinase (JNK), hypoxia inducible factor 1 alpha (HIF-1alpha), nuclear p65, and total p65 were assessed with western blotting. Apoptosis was identified by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling. RANK-Fc was efficiently expressed in the liver. In comparison with the negative-control group, RANK-Fc reduced nuclear factor kappa B (NF-kappaB) p65 nuclear translocation, JNK phosphorylation, and HIF-1alpha expression during I/R. RANK-Fc effectively suppressed proinflammatory cytokine expression. The results indicated that RANK-Fc could protect against hepatic I/R injury in mice at least in part via the inhibition of the proinflammatory NF-kappaB pathway as well as proapoptotic JNK and HIF-1alpha pathway activation.
Shi
Jian
J
Liver Transplantation Center, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, People's Republic of China.
Shao
Wenyu
W
Yang
Dongchang
D
Zhao
Liang
L
Deng
Lei
L
Wang
Xuehao
X
Sun
Beicheng
B
D000818:Animals; D017209:Apoptosis; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D015316:Genetic Therapy / Q000379:methods*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008107:Liver Diseases / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism; D010957:Plasmids / Q000493:pharmacokinetics; D053245:RANK Ligand / Q000378:metabolism; D053246:Receptor Activator of Nuclear Factor-kappa B / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D015427:Reperfusion Injury / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D015398:Signal Transduction / Q000502:physiology; D014162:Transfection / Q000379:methods*
D016207:Cytokines; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D016328:NF-kappa B; D053245:RANK Ligand; C517736:Rank-Fc; D053246:Receptor Activator of Nuclear Factor-kappa B; D011993:Recombinant Fusion Proteins; C505687:Tnfrsf11a protein, mouse; C505689:Tnfsf11 protein, mouse; D048031:JNK Mitogen-Activated Protein Kinases