Premium
Hydrodynamics‐based transfection of plasmid encoding receptor activator for nuclear factor kappa B–Fc protects against hepatic ischemia/reperfusion injury in mice
Author(s) -
Shi Jian,
Shao Wenyu,
Yang Dongchang,
Zhao Liang,
Deng Lei,
Wang Xuehao,
Sun Beicheng
Publication year - 2010
Publication title -
liver transplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.814
H-Index - 150
eISSN - 1527-6473
pISSN - 1527-6465
DOI - 10.1002/lt.22030
Subject(s) - terminal deoxynucleotidyl transferase , tunel assay , reperfusion injury , microbiology and biotechnology , apoptosis , medicine , kinase , activator (genetics) , biology , receptor , andrology , ischemia , biochemistry
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 1β 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‐1α), 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‐κB) p65 nuclear translocation, JNK phosphorylation, and HIF‐1α 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‐κB pathway as well as proapoptotic JNK and HIF‐1α pathway activation. Liver Transpl 16:611‐620, 2010. © 2010 AASLD.