Home In Vivo Proton Magnetic Resonance Spectroscopy of Hepatic Ischemia/Reperfusion Injury in an Experimental Model
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In Vivo Proton Magnetic Resonance Spectroscopy of Hepatic Ischemia/Reperfusion Injury in an Experimental Model

Rationale and Objectives

Hepatic ischemia/reperfusion injury (IRI) occurs during certain hepatobiliary surgeries, hemorrhagic shock, and veno-occlusive disease. Biochemical changes caused by hepatic IRI lead to hepatocellular remodeling, including cellular regeneration or irreversible apoptosis. This study aims to characterize and monitor the metabolic changes in hepatic IRI using proton magnetic resonance spectroscopy ( 1 H MRS).

Materials and Methods

Sprague-Dawley rats ( n = 8) were scanned with 1 H MRS using 5.0 × 5.0 × 5.0 mm 3 voxel over a homogeneous liver parenchyma at 7 Tesla with a respiratory-gated point-resolved spectroscopy sequence at 1 day before, 6 hours, 1 day, and 1 week after 30 minutes total hepatic IRI. Signal integral ratios of choline-containing compounds (CCC), glycogen and glucose complex (Glyu), methylene proton ((-CH 2 -) n ), and methene proton (-CH=CH-) to lipid (integral sum of methyl proton (-CH 3 ), (-CH 2 -) n and -CH=CH-) were quantified by areas under peaks longitudinally.

Results

The CCC-to-lipid and Glyu-to-lipid ratios at 6 hours after IRI were significantly higher than those at 1 day before, 1 day, and 1 week after injury. The (-CH 2 -) n -to-lipid, and -CH=CH-to-lipid ratios showed no significant differences over different time points. Hepatocellular regeneration was observed at 6 hours after IRI in histology with immunohistochemical technique.

Conclusions

Changes in CCC-to-lipid and Glyu-to-lipid ratios likely reflect the hepatocellular remodeling and impaired glucose utilization upon hepatic IRI, respectively. The experimental findings in the current study demonstrated that 1 H MRS is a valuable tool for characterizing either global or regional metabolic changes in liver noninvasively and longitudinally. Such capability has the potential to lead to early diagnosis and detection of impaired liver function.

Hepatic ischemia/reperfusion injury (IRI) induced by vascular complications contributes to early organ failure and can lead to acute and chronic rejection after liver transplantation . Hepatic IRI also occurs during certain hepatobiliary resectional surgeries, hemorrhagic shock, and veno-occlusive disease. IRI is a major cause of acute liver failure, which is associated with high morbidity and mortality . Cellular damage can be induced not only during ischemia but also in reperfusion that follows, resulting in both local and systemic organ dysfunction . Biochemical changes caused by hepatic IRI lead to hepatocellular remodeling, including cellular regeneration or irreversible programmed cell death . Early diagnosis and detection of impaired liver function is vital for early and effective therapeutic interventions and thus prevents its progression to liver failure. Through assessing liver function, serology screenings of liver-specific transaminases (aspartate aminotransferase and alanine aminotransferase) have been widely used to monitor hepatic IRI .

Metabolic changes of liver can provide information and improve the characterization of liver abnormalities . Monitoring of liver metabolism through a microdialysis catheter placed in the liver tissue revealed profound metabolic changes during and after IRI, leading to potential evaluation of ischemic preconditioning and clinical application . However, microdialysis technique is invasive in nature because implantation of microdialysis probe needs to be accompanied by surgical procedures . Recently, proton magnetic resonance spectroscopy ( 1 H MRS) has been increasingly employed to investigate liver metabolism in vivo noninvasively in various diseases . Biochemical changes occurred during hepatic IRI were also investigated in liver extracts . However, in vivo and serial study of hepatic IRI model with such MRS technique has been limited.

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Materials and methods

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Animal preparation

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MR spectroscopy

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Data and statistical analysis

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Histology

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Results

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Figure 1, (a) Illustration of the voxel placement (solid-line box) in the adult rat liver for proton magnetic resonance spectroscopy ( 1 H MRS) at 7 Tesla. (b) Typical liver 1 H MRS spectra from the same animal at 1 day before, 6 hours, 1 day, and 1 week after 30 minutes hepatic ischemia/reperfusion injury (IRI). Animals at 6 hours after hepatic IRI consistently showed markedly increased CCC and Glyu level. -CH 3 : methyl proton; (-CH 2 -) n : methylene proton; -CH=CH-CH 2 -: allylic proton; Glx: glutamine and glutamate complex; =CH-CH 2 -CH=: diallylic proton; CCC: choline-containing compounds; Glyu: glycogen and glucose complex; -CH=CH-: methene proton.

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Figure 2, CCC-to-lipid, Glyu-to-lipid (-CH 2 -) n -to-lipid, and -CH=CH-to-lipid ratios from all animals ( n = 8) at 1 day before injury, 6 hours, 1 day, and 1 week after hepatic IRI. Lipid was defined as the integral sum of -CH 3 , (-CH 2 -) n and -CH=CH-. One-way analysis of variance with Tukey’s multiple comparison test was performed with ∗ for P < .05, ∗∗ for P < .01, ∗∗∗ for P < .001 and n.s. for insignificance.

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Figure 3, Typical proliferating cell nuclear antigen (PCNA) staining observed in liver: (a) normal rat; (b) 6 hour; (c) 24 hours; and (d) 1 week after hepatic ischemia/reperfusion injury (IRI), with proliferating cells indicated by the incorporation of PCNA ( red arrows ). Magnified views of the corresponding blue boxes in left column (×100) are shown in right column (×400).

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Discussions

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