Nisanne Ghonem1, Junichi Yoshida2, Noriko Murase2, Stephen C Strom3, Raman Venkataramanan4. 1. University of Pittsburgh School of Pharmacy, Department of Pharmaceutical Sciences, Pittsburgh, PA, USA. 2. School of Medicine, Department of Surgery, Thomas E. Starzl Transplantation Institute, Pittsburgh, PA, USA. 3. Professor, Cellular Transplantation, Division of Pathology, Department of Laboratory Medicine, Karolinska Institutet and Hospital, Stockholm 141-86, Sweden. 4. University of Pittsburgh School of Pharmacy, Department of Pharmaceutical Sciences, Pittsburgh, PA, USA ; School of Medicine, Department of Surgery, Thomas E. Starzl Transplantation Institute, Pittsburgh, PA, USA.
Abstract
BACKGROUND: Cytochrome P450 (CYP450) activity is an important indicator of liver graft function. CYP450 activity is altered by pro-inflammatory cytokines, which are associated with ischemia-reperfusion (I/R) injury during orthotopic liver transplantation (OLT). Treprostinil, an FDA-approved prostacyclin analog, ameliorated cold I/R injury during rat OLT. We hypothesized that treprostinil would improve CYP450 activity in liver graft during cold I/R injury post-OLT. METHODS: OLT was performed in syngeneic male Lewis rats with 18 h graft preservation in cold UW solution. Donor and recipients received treprostinil (100 ng/kg/min) or matching placebo for 24 h before and up to 48 h post-OLT. Liver graft mRNA and protein expression of CYP450 isoforms were analyzed by qRT-PCR and Western blot analysis, respectively. The formation rates of 1-hydroxymidazolam and 6β-hydroxytestosterone, 6-hydroxychlorzoxazone, 2α- and 16α-hydroxytestosterone in liver graft microsomes served as markers for CYP3A, CYP2E1, and CYP2C11 activity, respectively, and were measured by LC-MS. RESULTS: Treprostinil significantly decreased serum ALT and AST levels at 6-48 h after OLT, compared to placebo. The expressions of TNFα and IFNγ mRNA in the liver graft were significantly inhibited in the treprostinil-treated group at 1 h post-reperfusion. Treprostinil restored CYP2E1 protein expression to that of normal liver and significantly improved CYP3A activity to more than two-fold of placebo early post-OLT. CONCLUSIONS: Treprostinil significantly ameliorated hepatic injury, reduced expression of pro-inflammatory cytokines, and improved CYP450 activity in liver graft early post-OLT. These findings suggest that treprostinil has the potential to serve as a therapeutic option to protect liver graft function against I/R injury during clinical OLT.
BACKGROUND:Cytochrome P450 (CYP450) activity is an important indicator of liver graft function. CYP450 activity is altered by pro-inflammatory cytokines, which are associated with ischemia-reperfusion (I/R) injury during orthotopic liver transplantation (OLT). Treprostinil, an FDA-approved prostacyclin analog, ameliorated cold I/R injury during rat OLT. We hypothesized that treprostinil would improve CYP450 activity in liver graft during cold I/R injury post-OLT. METHODS: OLT was performed in syngeneic male Lewis rats with 18 h graft preservation in cold UW solution. Donor and recipients received treprostinil (100 ng/kg/min) or matching placebo for 24 h before and up to 48 h post-OLT. Liver graft mRNA and protein expression of CYP450 isoforms were analyzed by qRT-PCR and Western blot analysis, respectively. The formation rates of 1-hydroxymidazolam and 6β-hydroxytestosterone, 6-hydroxychlorzoxazone, 2α- and 16α-hydroxytestosterone in liver graft microsomes served as markers for CYP3A, CYP2E1, and CYP2C11 activity, respectively, and were measured by LC-MS. RESULTS:Treprostinil significantly decreased serum ALT and AST levels at 6-48 h after OLT, compared to placebo. The expressions of TNFα and IFNγ mRNA in the liver graft were significantly inhibited in the treprostinil-treated group at 1 h post-reperfusion. Treprostinil restored CYP2E1 protein expression to that of normal liver and significantly improved CYP3A activity to more than two-fold of placebo early post-OLT. CONCLUSIONS:Treprostinil significantly ameliorated hepatic injury, reduced expression of pro-inflammatory cytokines, and improved CYP450 activity in liver graft early post-OLT. These findings suggest that treprostinil has the potential to serve as a therapeutic option to protect liver graft function against I/R injury during clinical OLT.
Entities:
Keywords:
1-OH MDZ, 1-hydroxymidazolam; 16α-OH TST, 16α-hydroxytestosterone; 2α-OH TST, 2α-hydroxytestosterone; 6-OH CZN, 6-hydroxychlorzoxazone; 6β-OH TST, 6β-hydroxytestosterone; ALT, alanine aminotransferase; AST, aspartate aminotransferase; AUC, area under the time-concentration curves; CYP450, cytochrome P450; CZN, chlorzoxazone; HPLC-mass spectrometry; I/R, ischemia-reperfusion; IFN-γ, interferon gamma; IL, interleukin; Ischemia-reperfusion injury; MDZ, midazolam; NF-κB, nuclear factor-kappa B; NL, normal liver; OLT, orthotopic liver transplantation; PG, prostaglandin; PGI2, prostacyclin; TNF-α, tumor necrosis factor alpha; TST, testosterone; UW, University of Wisconsin; cytokines; drug metabolism; mRNA, messenger RNA; prostacyclin
Authors: N Ghonem; J Yoshida; D B Stolz; A Humar; T E Starzl; N Murase; R Venkataramanan Journal: Am J Transplant Date: 2011-06-10 Impact factor: 8.086
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