Assessing steatotic liver function after ischemia-reperfusion injury by in vivo multiphoton imaging of fluorescein disposition.
Ischemia-reperfusion injury, a common complication during liver surgery where steatotic livers are more prone to the injury, may become more prevalent in the growing obese population. This study characterizes liver morphology toward understanding changes in subcellular function in steatotic livers exposed to ischemia-reperfusion injury through quantitative description of fluorescein distribution obtained by minimally invasive in vivo multiphoton microscopy using a physiologic pharmacokinetic model. Rats were fed a high-fat diet for 7 days to induce liver steatosis. Partial ischemia was induced after reperfusion for 4 hours, when fluorescein (10 mg/kg) was injected intravenously. Liver images, bile, and blood were collected up to 180 minutes after injection. Ischemia-reperfusion injury was associated with an increase in alanine transaminase levels and apoptosis. In addition, steatosis featured lipid droplets and an increase in fluorescein-associated fluorescence observed in hepatocytes via multiphoton imaging. Analysis of the hepatic concentration-time profiles has suggested that the steatosis-induced increase in fluorescein-associated fluorescence mainly arises by inducing hepatic fluorescein metabolism. The combination of ischemia-reperfusion with steatosis exacerbates these effects further. This was confirmed by fluorescence lifetime imaging microscopy showing a decreased average fluorescence lifetime of the liver, which is indicative of an increased production of the metabolite. Our results show the potential of noninvasive dye imaging for improving our understanding of liver disease induced by subcellular changes in vivo, providing further quantitative measures of metabolic and biliary liver function, and hence extending the qualitative liver function tests now available.
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.).
Camilla A
CA
Thorling
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.).
Lu
L
Jin
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.).
Michael
M
Weiss
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.).
Darrell
D
Crawford
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.).
Xin
X
Liu
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.).
Frank J
FJ
Burczynski
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.).
David
D
Liu
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.).
Haolu
H
Wang
School of Pharmacy and Biomedical Sciences, University of South Australia, Adelaide, Australia (C.A.T., M.S.R.); School of Medicine, University of Queensland, Wolloongabba, Queensland, Australia (C.A.T., M.W., D.C., X.L., D.L., H.W., M.S.R.); Nanjing Drum Tower Hospital Affiliated to the Medical School of Nanjing University, Nanjing, People's Republic of China (L.J.); Department of Pharmacology, Martin Luther University Halle-Wittenberg, Halle, Germany (M.W.); and Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada (F.J.B.) m.roberts@uq.edu.au.
Michael S
MS
Roberts
D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D001646:Bile / Q000378:metabolism; D003952:Diagnostic Imaging / Q000379:methods; D059305:Diet, High-Fat / Q000009:adverse effects; D005234:Fatty Liver / Q000378:metabolism* / Q000503:physiopathology*; D019793:Fluorescein / Q000378:metabolism*; D008099:Liver / Q000378:metabolism* / Q000503:physiopathology*; D008111:Liver Function Tests / Q000379:methods; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000378:metabolism* / Q000503:physiopathology*
D000410:Alanine Transaminase; D019793:Fluorescein