Erythrocytes with T-state-stabilized hemoglobin as a therapeutic tool for postischemic liver dysfunction.
This study aimed to examine if T-state stabilization of hemoglobin in erythrocytes could protect against postischemic organ injury. Human erythrocytes containing three different states of Hb allostery were prepared: control Hb (hRBC), CO-Hb that is stabilized under R-state with the 6-coordinated prosthetic heme (CO-hRBC), and alpha-NO-deoxyHb stabilized under T-state (alpha-NO-hRBC). To prepare alpha-NO-RBC, deoxygenated RBC was treated with FK409, a thiol-free NO donor, at its half molar concentration to that of Hb; this procedure resulted in the 5-coordinated NO binding on the alpha-subunit heme, as judged by electron spin resonance spectrometry. Rats were subject to 20 min systemic hemorrhage to maintain mean arterial pressure at 40 mm Hg, and reperfused with one of hRBCs. This protocol for ischemia, followed by 60 min reperfusion with physiological saline, caused modest metabolic acidosis and cholestasis. Administration of hRBC or COhRBC significantly attenuated cholestasis and improved acidosis. Rats treated with alpha-NO-hRBC exhibited greater recovery of metabolic acidosis and bile excretion than those treated with hRBC or CO-hRBC, displaying the best outcome of local oxygen utilization in hepatic lobules. Half-life time of alpha-NO-RBC administered in vivo was approximately 60 min. These results suggest that T-state Hb stabilization by NO serves as a stratagem to treat postischemic organ dysfunction.
Suganuma
Kazuhiro
K
Department of Biochemistry and Integrative Medical Biology, University of Pennsylvania, Philadelphia, USA.
Tsukada
Kosuke
K
Kashiba
Misato
M
Tsuneshige
Antonio
A
Furukawa
Toshiharu
T
Kubota
Tetsuro
T
Goda
Nobuhito
N
Kitajima
Masaki
M
Yonetani
Takashi
T
Suematsu
Makoto
M
D000138:Acidosis / Q000097:blood / Q000628:therapy; D000818:Animals; D001646:Bile / Q000378:metabolism; D001794:Blood Pressure / Q000502:physiology; D002248:Carbon Monoxide / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D017707:Erythrocyte Transfusion; D004912:Erythrocytes / Q000737:chemistry; D006339:Heart Rate / Q000502:physiology; D006454:Hemoglobins / Q000737:chemistry / Q000493:pharmacokinetics / Q000627:therapeutic use*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008107:Liver Diseases / Q000378:metabolism / Q000628:therapy*; D008297:Male; D020030:Nitric Oxide Donors / Q000737:chemistry; D009574:Nitro Compounds / Q000737:chemistry; D010100:Oxygen / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012039:Regional Blood Flow / Q000502:physiology; D015427:Reperfusion Injury / Q000628:therapy*
D006454:Hemoglobins; D020030:Nitric Oxide Donors; D009574:Nitro Compounds; D002248:Carbon Monoxide; C100003:deoxyhemoglobin; C061872:FK 409; D010100:Oxygen