Carbon Monoxide–Bound Red Blood Cells Protect Red Blood Cell Transfusion-Induced Hepatic Cytochrome P450 Impairment in Hemorrhagic-Shock Rats

Abstract

Red blood cell (RBC) transfusions for massive hemorrhage induce systemic ischemic-reperfusion and influence the disposition and pharmacological activity of drugs as a result of a reduction in the level of expression and activity of cytochrome P450s (P450). It was reported that, when organ-preserving solutions are exposed to carbon monoxide (CO), the treatment was effective in suppressing the postreperfusion reduction in renal P450 levels in cases of kidney transplantation. Therefore, we hypothesized that transfusions with RBC that contain bound CO (CO-RBC) would protect the hepatic level of rat P450 during a massive hemorrhage, compared with plasma expanders and RBC resuscitation. To achieve this, we created 40% hemorrhagic-shock model rats, followed by resuscitation, with use of recombinant human serum albumin, RBCs, and CO-RBCs. At 1 hour after resuscitation, the expressions of hepatic P450 isoforms (1A2, 2C11, 2E1, and 3A2) were significantly decreased in the RBC resuscitation group, compared with the sham group. Such alterations in hepatic P450 significantly resulted in an increase in the plasma concentrations of substrate drugs (caffeine [1A2], tolbutamide [2C11], chlorzoxazone [2E1], and midazolam [3A2]) for each P450 isoform, and thus, the hypnotic action of midazolam could be significantly prolonged. Of interest, the reductions in hepatic P450 activity observed in the RBC group were significantly suppressed by CO-RBC resuscitation, and consequently, the pharmacokinetics of substrate drugs and the pharmacological action of midazolam remained at levels similar to those under sham conditions. These results indicate that CO-RBC resuscitation has considerable potential in terms of achieving safe and useful drug therapy during massive hemorrhages.

Kupffer cell inactivation by carbon monoxide bound to red blood cells preserves hepatic cytochrome P450 via anti-oxidant and anti-inflammatory effects exerted through the HMGB1/TLR-4 pathway during resuscitation from hemorrhagic shock

“…Each sample was separated by 12.5% polyacrylamide gel electrophoresis, and subsequently transferred to polyvinylide difluoride membranes by wet electroblotting. Bands were immunologically detected by the same procedure as our previous report [5].…”

“…The washed RBC resuscitative fluid was prepared from donor rats as described in a previous report [5,16], and the hemoglobin (Hb) concentration of the RBC resuscitative fluid was subsequently adjusted to 10 g/dL. CO-RBC resuscitative fluid was prepared by gently bubbling CO gas through the RBC resuscitative fluid.…”

“…Hemorrhagic shock model rats were prepared by removing 40% of the total blood volume according to our previous report [5,16]. Under these experimental conditions, the mean arterial pressure remained at less than 40 mm Hg for 30 min.…”

“…Blood samples were collected at multiple time points after injection of the midazolam (5, 15, 30 min, 1, 2, 4, 6, 12, and 24 hr) and the plasma was separated by centrifugation (3,000 g, 5 min, 4°C). The concentration of midazolam in plasma were determined by high-performance liquid chromatography according to a previous report [5].…”

“…In a previous study, we reported that carbon monoxide (CO)-bound RBC (CO-RBC) had the potential for use as a resuscitative fluid under conditions of massive hemorrhage and resuscitation that overcame the limitations associated with RBC transfusions [5]. In addition, in order to be as effective as RBC as a resuscitation fluid, CO-RBC resuscitation from hemorrhagic shock protected against the destruction of hepatic CYP compared to that by RBC.…”