Heme oxygenase-1 (HO-1) cleaves heme to form biliverdin, carbon monoxide (CO) and iron (Fe2+), which is used with 5-ALA. We have recently reported that 5-ALA with Fe2+ (5-ALA/Fe2+) can protect the kidney against I/R renal injury. In the present study we tried to investigate the hypothesis that 5-ALA/Fe2+ has a beneficial effect on acute I/R injury in mouse steatotic liver model. Methods: Male C57BL/6
mice were all fed with methionine and choline-defi-cient high fat (MCDHF) diet for 3 weeks to establish steatotic liver model, then randomized into 5 groups as follows: MCDHF diet (MCDHFD); MCDHF diet and saline treated before I/R (MCDHF I/R); MCDHF diet and 5-ALA/Fe2+ treated before IR (MCDHF+5-ALA/Fe2+ I/R). 5-ALA /Fe2+ was orally administrated 3 times at 48, 24 and 0.5 hr before ischemia. I/R liver injury was induced warm ischemia for 15min, followed by 1hr or 3hrs reperfusion in (1h) and (3h) BMN 673 solubility dmso group, respectively. Then, the liver and serum were examined. For in vitro study, inflammatory cytokines were measured by treated with or without 5-ALA/Fe2+
in LPS-stimulated RAW 264.7 cells. Results: Serum AST and ALT levels, thiobarbituric acid-reactive substances (TBARS) content in the liver, the area of necrosis in the liver, the number of TUNEL-positive cells and F4/80 positive macro-phages were significantly higher in both MCDHF I/R the (1h) and (3h) groups than the MCDHFD group, and were dramatically attenuated in MCDHF+5-ALA/Fe2+ both (1h) and (3h). Compared to MCDHF I/R (1h) and (3h) groups, inflammatory cytokine genes such as TNF-α, IL-6, osteopontin, INF-y, GDC-0068 ic50 iNOS were all markedly reduced by 5-ALA/Fe2+ treatment (p<0.05 respectively). Endogenous CO concentration in the steatotic liver was up-regulated
at 30 and 60 minutes after oral administration of 5-ALA/Fe2+. Moreover, HO-1 expression was significantly increased by treatment with 5-ALA/Fe2+e. In vitro study in RAW264.7 cells, 5-ALA/Fe2+ significantly diminished the expression of inflammatory cytokines, but induced HO-1 expression. Conclusion: These NADPH-cytochrome-c2 reductase results suggest that 5-ALA/Fe2+ noticeably protected I/R injury in mouse fatty liver model. We identify the protective effects of 5-ALA/Fe2+ by its anti-oxi-dant, anti-inflammatory and anti-apoptotic mechanisms through the generation of endogenous CO and up-regulation of HO-1 expression. Thus, 5-ALA/Fe2+ may be a promising candidate for a liver transplantation pretreatment. Disclosures: The following people have nothing to disclose: Shao-Wei Li, Terumi Takahara, Toshiro Sugiyama, Kazuhiro Tsukada, Tohru Tanaka, Xiao-Kang Li Background & Aim: Earlier therapeutic intervention in abnormal glucose metabolism may prevent the progression of nonalcoholic fatty liver disease (NAFLD), since insulin resistance is a risk factor for disease progression in NAFLD.