BOLD quantified renal pO2 is sensitive to pharmacological challenges in rats.

PURPOSE: Blood oxygen level-dependent (BOLD) MRI has been effectively used to monitor changes in renal oxygenation. However, R2* (or T2*) is not specific to blood oxygenation and is dependent on other factors. This study investigates the use of a statistical model that takes these factors into account and maps BOLD MRI measurements to blood pO2. METHODS: Spin echo and gradient echo images were obtained in six Sprague-Dawley rats and R2 and R2* maps were computed. Measurements were made at baseline, post-nitric oxide synthase inhibitor (L-NAME), and post-furosemide administration. A simulation of each region was performed to map R2' (computed as R2*-R2) to blood pO2. RESULTS: At baseline, blood pO2 in the outer medulla was 30.5 ± 1.2 mmHg and 51.9 ± 5.2 mmHg in the cortex, in agreement with previous invasive studies. Blood pO2 was found to decrease within the outer medulla following L-NAME (P < 0.05) and increase after furosemide (P < 0.05). Blood pO2 in the cortex increased following furosemide (P < 0.05). CONCLUSIONS: Model-derived blood pO2 is sensitive to pharmacological challenges, and baseline pO2 is comparable to literature values. Reporting pO2 instead of R2* could lead to a greater clinical impact of renal BOLD MRI and facilitate the identification of hypoxic regions. Magn Reson Med 78:297-302, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

Effect of iodinated contrast medium in diabetic rat kidneys as evaluated by blood-oxygenation-level-dependent magnetic resonance imaging and urinary neutrophil gelatinase-associated lipocalin.

OBJECTIVES: The objective of this study was to assess whether streptozotocin (STZ)-induced diabetic rats develop iodinated contrast-induced acute kidney injury. The intrarenal R2* (=1/T2*) was evaluated continuously before, during, and after contrast administration. Renal injury was confirmed using urinary neutrophil gelatinase-associated lipocalin measurements. MATERIALS AND METHODS: Six Sprague-Dawley rats were administered with STZ to induce diabetes (group 1). R2* was measured before, during, and after administration of iodixanol. R2* readings were sampled from 4 renal regions: inner medulla, inner stripe of outer medulla (ISOM), outer stripe of outer medulla, and cortex. Peak R2* and initial upslope of R2* increase after iodinated contrast were calculated. Data from 12 nondiabetic rats pretreated with nitric oxide synthase and prostaglandin inhibitors to induce susceptibility to contrast-induced acute kidney injury (pretreatment model) from a previous study were reanalyzed for peak R2* and initial upslope of R2* increase after contrast. Six of these animals received saline (group 2), and the other 6 received furosemide (group 3) before iodixanol. RESULTS: Peak R2* and initial upslope of R2* increase were used as blood-oxygenation-level-dependent response parameters. R2* in ISOM was comparable in all 3 groups before administration of furosemide/saline. Except for the furosemide group, ISOM showed a rapid increase in R2* immediately after contrast administration. Unlike the L-NAME- and indomethacin-treated groups, the diabetic group showed a quick reversal of R2* toward baseline measurements after contrast administration. Urinary neutrophil gelatinase-associated lipocalin indicated significant increase in diabetic rats 4 hours after contrast administration. The observed trends with peak R2* and initial upslope of R2* increase in renal ISOM were in agreement with those of urinary neutrophil gelatinase-associated lipocalin. CONCLUSIONS: The STZ-induced diabetic rat may be suitable for studying the effects of iodinated contrast on renal oxygenation status and may mimic human condition closer than the pretreatment model described before. The peak R2* value and initial upslope of R2* in ISOM appear to be effective magnetic resonance imaging markers to predict renal injury after administration of an iodinated contrast agent.

Sensitivity of arterial spin labeling perfusion MRI to pharmacologically induced perfusion changes in rat kidneys.

PURPOSE: To investigate whether arterial spin labeling (ASL) MRI is sensitive to changes by pharmacologically induced vasodilation and vasoconstriction in rat kidneys. MATERIALS AND METHODS: Changes in renal cortical blood flow in seven rats were induced by adenosine infusion (vasodilation) and L-NAME injection (vasoconstriction). All imaging studies were performed on a 3 Tesla scanner using a FAIR-TrueFISP sequence for the ASL implementation. The acquisition time for each ASL scan was 6 min. Cortical perfusion rates were calculated using regions of interest analysis, and the differences in perfusion rates during baseline, vasodilation, and vasoconstriction were compared and assessed for statistical significance. RESULTS: Compared with the baseline, an average of 94 mL/100 g/min increase and 157 mL/100 g/min decrease in cortical perfusion was observed following adenosine infusion and L-NAME administration, respectively. The changes in cortical perfusion were significant between baseline and vasodilation (P < 0.05), baseline and vasoconstriction (P < 0.01), and vasodilation and vasoconstriction (P < 0.01). CONCLUSION: ASL is sensitive to pharmacologically induced perfusion changes in rat kidneys at doses comparable to current use. The preliminary results suggest the feasibility of ASL for investigating renal blood flow in a variety of rodent models.

Efficacy of preventive interventions for iodinated contrast-induced acute kidney injury evaluated by intrarenal oxygenation as an early marker.

OBJECTIVE: The objective of this study was to evaluate the effects of potential renoprotective interventions such as the administration of N-acetylcysteine (NAC; antioxidant) and furosemide (diuretic) on intrarenal oxygenation as evaluated by blood oxygen level-dependent (BOLD) magnetic resonance imaging (MRI) in combination with urinary neutrophil gelatinase-associated lipocalin (NGAL) measurements. MATERIALS AND METHODS: Rats received nitric oxide synthase inhibitor L-NAME (10 mg/kg) and cyclooxygenase inhibitor indomethacin (10 mg/kg) to induce the risk for developing iodinated contrast-induced acute kidney injury before receiving one of the interventions: NAC, furosemide, or placebo. One of the 3 iodinated contrast agents (iohexol, ioxaglate, or iodixanol) was then administered (1600-mg organic iodine per kilogram body weight). Fifty-four Sprague-Dawley rats were allocated in a random order into 9 groups on the basis of the intervention and the contrast agent received.Blood-oxygen-level-dependent MRI-weighted images were acquired on a Siemens 3.0-T scanner using a multiple gradient recalled echo sequence at baseline, after L-NAME, indomethacin, interventions or placebo, and iodinated contrast agents. Data acquisition and analysis were performed in a blind fashion. R2* (=1/T2*) maps were generated inline on the scanner. A mixed-effects growth curve model with first-order autoregressive variance-covariance was used to analyze the temporal data. Urinary NGAL, a marker of acute kidney injury, was measured at baseline, 2 and 4 hours after the contrast injection. RESULTS: Compared with the placebo-treated rats, those treated with furosemide showed a significantly lower rate of increase in R2* (P < 0.05) in the renal inner stripe of the outer medulla. The rats treated with NAC showed a lower rate of increase in R2* compared with the controls, but the difference did not reach statistical significance. Urinary NGAL showed little to no increase in R2* after administration of iodixanol in the rats pretreated with furosemide but demonstrated significant increase in the rats pretreated with NAC or placebo (P < 0.05). CONCLUSIONS: This is the first study to evaluate the effects of interventions to mitigate the deleterious effects of contrast media using BOLD MRI. The rate of increase in R2* after administration of iodinated contrast is associated with acute renal injury as evaluated by NGAL. Further studies are warranted to determine the optimum dose of furosemide and NAC for mitigating the ill effects of contrast media. Because NGAL has been shown to be useful in humans to document iodinated contrast-induced acute kidney injury, the method presented in this study using BOLD MRI and NGAL measurements can be translated to humans.

Evaluation of intrarenal oxygenation in iodinated contrast-induced acute kidney injury-susceptible rats by blood oxygen level-dependent magnetic resonance imaging.

OBJECTIVES: The objectives of this study were to evaluate differences in intrarenal oxygenation as assessed by blood oxygen level-dependent (BOLD) magnetic resonance imaging in contrast-induced acute kidney injury (CIAKI)-susceptible rats when using 4 contrast media with different physicochemical properties and to demonstrate the feasibility of acquiring urinary neutrophil gelatinase-associated lipocalin (NGAL) levels as a marker of CIAKI in this model. MATERIALS AND METHODS: Our institutional animal care and use committee approved the study. Sixty-six Sprague-Dawley rats were divided into CIAKI-susceptible groups (received nitric oxide synthase inhibitor N-nitro-L-arginine methyl ester [10 mg/kg] and cycloxygenase inhibitor indomethacin [10mg/kg]) and control groups (received saline instead). One of the 4 iodinated contrast agents (iothalamate, iohexol, ioxaglate, or iodixanol) was then administered (1600-mg organic iodine per kilogram of body weight). Multiple blood oxygen level-dependent magnetic resonance images were acquired on a Siemens 3.0-T scanner using a multiple gradient recalled echo sequence at baseline, after N-nitro-L-arginine methyl ester (or saline), indomethacin (or saline), and iodinated contrast agent (or placebo). R2* (R2*=1/T2*) maps were generated inline on the scanner. A mixed-effects growth curve model with first-order autoregressive variance-covariance was used to analyze the temporal data. Urinary NGAL, a marker of kidney injury (unlike serum creatinine), was measured 4 hours after contrast injection in the 2 subgroups. RESULTS: Differences in blood oxygen level-dependent magnetic resonance imaging results between the contrast media were observed in all 4 renal regions. However, the inner stripe of the outer medulla (ISOM) showed the most pronounced changes in the CIAKI-susceptible group and R2* increased significantly (P<0.01) over time with all 4 contrast media. In the control groups, only iodixanol showed an increase in R2* (P<0.05) over time. There was an agreement between increases in NGAL and R2* values in ISOM. CONCLUSIONS: In rats susceptible to CIAKI, those receiving contrast media had significant increases in R2* in renal ISOM compared with those receiving placebo. The agreement between NGAL and R2* values in the ISOM suggests that the observed immediate increase in R2* after contrast injection may be the earliest biomarker of renal injury. Further studies are necessary to establish threshold values of R2* associated with acute kidney injury and address the specificity of R2* to renal oxygenation status.

Longitudinal changes in MRI markers in a reversible unilateral ureteral obstruction mouse model: preliminary experience.

PURPOSE: To evaluate longitudinal changes in renal oxygenation and diffusion measurements in a model of reversible unilateral ureteral obstruction (rUUO) which has been shown to induce chronic renal functional deficits in a strain dependent way. C57BL/6 mice show higher degree of functional deficit compared with BALB/c mice. Because hypoxia and development of fibrosis are associated with chronic kidney diseases and are responsible for progression, we hypothesized that MRI measurements would be able to monitor the longitudinal changes in this model and will show strain dependent differences in response. Here blood oxygenation level dependent (BOLD) and diffusion MRI measurements were performed at three time points over a 30 day period in mice with rUUO. MATERIALS AND METHODS: The studies were performed on a 4.7T scanner with the mice anesthetized with isoflurane before UUO, 2 and 28 days postrelease of 6 days of obstruction. RESULTS: We found at the early time point (∼2 days after releasing the obstruction), the relative oxygenation in C57Bl/6 mice were lower compared with BALB/c. Diffusion measurements were lower at this time point and reached statistical significance in BALB/c CONCLUSION: These methods may prove valuable in better understanding the natural progression of kidney diseases and in evaluating novel interventions to limit progression.

Intrarenal oxygenation by blood oxygenation level-dependent MRI in contrast nephropathy model: effect of the viscosity and dose.

PURPOSE: To compare the effects of osmolality versus viscosity of radio-contrast media on intra-renal oxygenation as determined by blood oxygenation level-dependent (BOLD) MRI in a model of contrast induced nephropathy (CIN). MATERIALS AND METHODS: Twenty-four Sprague-Dawley rats were divided into five groups. Nitric oxide synthase inhibitor L-NAME (10 mg/kg), cyclooxygenase inhibitor indomethacin (10 mg/kg), or saline, and radio-contrast iodixanol (high viscosity, 784 or 1600 mg I/kg) or iothalamate (high osmolality, 1600 mg I/kg) were administered. BOLD MRI images were acquired on Siemens 3 Tesla (T) scanner using a multiple gradient recalled echo sequence at baseline, following L-NAME (or saline), indomethacin (or saline), and radio-contrast agents. R2* (=1/T2*) was used as the BOLD MRI parameter in renal medulla and cortex. Mixed-effects models with first order auto-regressive variance-covariance models were used to analyze the data. RESULTS: The magnitude of change in medullary R2* (MR2*) with same dose of iodine was larger with iodixanol compared with iothalalmate both in pretreated groups (303% versus 225.6%, < 0.01) and the control group (191.6% versus -1.8%, P < 0.01). The MR2* change in high dose iodixanol was approximately twice compared with the low dose (303% versus 133%, P < 0.01). CONCLUSION: The viscosity of radio-contrast seems to play a more significant role than osmolality in terms of renal oxygenation changes as evaluated by BOLD MRI. Additionally, iodixanol induced a dose-dependent increase in renal medullary hypoxia.

Renal oxygenation changes during water loading as evaluated by BOLD MRI: effect of NOS inhibition.

PURPOSE: To demonstrate a possible role for endogenous release of nitric oxide in determining the response of water loading on intrarenal oxygenation as evaluated by blood oxygenation level-dependent (BOLD) magnetic resonance imaging (MRI). MATERIALS AND METHODS: Twelve Sprague Dawley rats (weight 344.9 ± 40.6 g) were equally divided into two groups, A and B. Water loading was implemented by continuous infusion of hypotonic saline containing glucose (0.25% NaCl, 0.5% glucose). Rats in group A were subject to water loading alone, while group B rats were dosed with N-nitro-L-arginine methyl ester, (L-NAME) (10.0 mg/kg) prior to water loading. T(2) *-weighted images of the kidneys were obtained on a Siemens 3T Verio MRI scanner using a multiple gradient recalled echo (mGRE) sequence. RESULTS: Consistent with previous reports, group A exhibited a significant decrease in medullary R(2) * during water loading (40.64 ± 1.10 s(-1) to 34.68 ± 1.49 s(-1) , P < 0.05). On the other hand, in group B there was no decrease in R(2) * during water loading (48.11 ± 2.38 s(-1) to 51.06 ± 2.18 s(-1) ). The increased prewater loading R(2) * is due to the pretreatment with L-NAME (40.82 ± 3.23 s(-1) to 48.11 ± 2.38 s(-1) , P < 0.05). CONCLUSION: Our data suggest for the first time a role for endogenous nitric oxide in determining the response of renal medullary oxygenation to water loading.

Vascular perfused segments of human intestine as a tool for drug absorption.

Blood-based vascular perfusion of isolated segments of human jejunum was developed as a tool for drug absorption studies before clinical trials. Acceptance criteria for viable human gut preparations included stable blood flow, arterial pressure, glucose utilization, active peristalsis, oxygen uptake, less than 3% absorption of a 70,000 mol. wt. dextran, and a ratio of first-order absorption rate constants (k(a)) of antipyrine to terbutaline of > or =1.4. Mannitol absorption was less than that of antipyrine but larger than that of terbutaline and could not be used as a negative control in absorption studies with human intestine. In separate perfusions (n = 3) a cassette of nine drugs was administered into the gut lumen, and the net absorption of each drug into the circulation was measured over 75 min. Using the mean values of k(a), the test compounds could be ranked into four groups: group 1: sulfasalazine and furosemide, k(a) = 3.9 to 4.0 x 10(-3) min(-1); group 2: cimetidine, timolol, nadolol, and ranitidine, k(a) = 6.4 to 8.3 x 10(-3) min(-1); group 3: atenolol and metoprolol, k(a) = 9.6 x 10(-3) min(-1); and group 4: theophylline, k(a) = 17.5 x 10(-3) min(-1). The rationale for evaluating yet another oral absorption system was as follows: first, a human gut segment with an intact vascular system is the closest system available to a clinical trial without performing one; and second, the data generated would be a direct measure of net drug transport from the gut lumen into the vascular circulation under near physiological conditions, which is not possible in models lacking a blood supply.