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Liver Enzyme Levels and Hepatic Iron Content in Fatty Liver

Rationale and Objectives

Existing evidence suggests potential contribution of iron in pathogenesis of nonalcoholic fatty liver disease (NAFLD). We aimed to investigate whether hepatic iron content correlates with liver enzyme levels in NAFLD using a noninvasive magnetic resonance imaging (MRI) technique.

Materials and Methods

Subjects from Golestan Cohort Study were randomly selected. Diagnosis of NAFLD was made by combination of ultrasound and MRI. Subjects with NAFLD were divided into two groups with high (H-NAFLD) and low (L-NAFLD) enzyme level according to 95th percentile of alanine aminotransferase (ALT) value in normal population. Quantitative T2* maps of entire cross-sectional area of liver were calculated on pixel-by-pixel basis using a semiautomated software.

Results

A total of 207 subjects were enrolled. Mean T2* values were significantly lower in NAFLD group than controls ( P < .001) indicating higher iron content. Male subjects with H-NAFLD had statistically lower T2* values than those with L-NAFLD in multivariate analysis (odds ratio, 0.74; 95% confidence interval [CI], 0.58–0.95), whereas this was not observed in women. Unlike women, there was significant negative correlation between ALT levels and T2* values in men with H-NAFLD ( r = −0.66, P = .01). Every 1-millisecond decrement in T2* value was associated with 6.37 IU/L increase in ALT level (95% CI, 1.8–10.9, P = .01) in men with H-NAFLD.

Conclusions

Higher hepatic iron in men with H-NAFLD, estimated by T2* mapping, may support the role of iron in possible progression of simple steatosis to nonalcoholic steatohepatitis. Lack of such correlation in women could be attributed to relatively lower iron storage or other mechanisms rather than iron.

Nonalcoholic fatty liver disease (NAFLD) is a common cause of chronic liver disease, which is strongly associated with features of metabolic syndrome . Nonalcoholic steatohepatitis (NASH) is the severe form of NAFLD, which has a multifactorial etiologic model. Excess iron deposition has been proposed as one of the potential contributing factors in pathogenesis of NASH, which can complicate simple steatosis via the production of reactive oxygen species (ROS) . Iron may also have a role in mechanisms other than ROS formation including altered insulin signaling and lipid metabolism. Consequently, iron may contribute not only to NASH progression but also to the initial development of steatosis .

A number of studies have investigated the relationship between NAFLD and liver iron by demonstrating the presence of stainable hepatic iron deposit . However the results of these studies, which were obtained from specimens of liver biopsy, are conflicting as some support the association of hepatic iron deposition with more advanced NASH , whereas others do not . Although biopsy is the gold standard for evaluation of liver disease, it is an invasive procedure, which is subject to sampling error or nonhomogeneous distribution of liver injuries . Moreover, it cannot be used in population-based studies for both ethical and practical reasons.

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Materials and methods

Study Design and Subjects

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Ultrasound

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Magnetic Resonance Imaging

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Image Analysis

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Figure 1, Transverse T1-weighted in-phase (a) and opposed-phase (b) images of the liver in a 56-year-old man with nonalcoholic fatty liver disease. (c) Measurement of relative signal intensity loss using a home-made software by drawing six regions of interest (ROIs) in liver (four in right and two in left lobes) and three ROIs in spleen at each slice on in-phase image which was repeated for two other slices. The same pixels were automatically found on corresponding opposed-phase images. Relative signal intensity loss of the liver in opposed-phase images according to the average signal intensity of 18 ROIs in liver and 9 ROIs in spleen (reference organ) was calculated as about 60% suggestive of considerable steatosis.

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Figure 2, Transverse multiecho T2*-weighted images of a 54-year-old female subject (a) . Using a semiautomatic program, the entire cross-sectional area of the liver was selected manually in each slice and then the major vessels and artifacts were excluded (b) . The segmented liver image was obtained (c) . T2* color map of the whole liver was generated on a pixel-by-pixel basis by relaxometry method which revealed a mean T2* value of 19.6 milliseconds (d) .

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Definitions

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Statistical Analysis

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Results

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Table 1

Demographic Characteristics and Laboratory Findings of Participants in Three Studied Groups

Characteristic Control L-NAFLD H-NAFLD_N_ 97 85 25 F/M 51/46 47/38 11/14 Age range (years) 50–77 50–72 50–70 Age (years; mean ± SD) 57.87 ± 6.52 56.75 ± 5.61 54.84 ± 4.48 BMI range 19.03–39.94 21.55–37.39 25.68–35.83 BMI (mean ± SD) 27.64 ± 4.12 29.78 ± 3.44 30.01 ± 2.73 ALT (IU/mL; mean ± SD) 20.79 ± 5.12 24.76 ± 7.33 65.64 ± 22.57 AST (IU/mL; mean ± SD) 20.20 ± 6.66 20.75 ± 7.26 38.79 ± 20.05 T2* (ms; mean ± SD) 23.00 ± 3.50 20.50 ± 4.31 18.78 ± 3.66 Ferritin (ng/mL; mean ± SD) 81.76 ± 62.58 108.05 ± 52.95 128.92 ± 64.42 Hemoglobin (mg/dL; mean ± SD) 12.96 ± 1.43 13.41 ± 1.69 13.83 ± 1.50

ALT, alanine aminotransferase; AST, aspartate aminotransferase; BMI, body mass index; H-NAFLD, high enzyme level NAFLD; L-NAFLD, low enzyme level NAFLD; NAFLD, nonalcoholic fatty liver disease; SD, standard deviation.

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Univariate Analysis

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Table 2

Univariate Analysis for Association of Demographics, T2* Value, and Risk Factors of Metabolic Syndrome with Low and High Enzyme Level NAFLD in Men and Women

Variable Control L-NAFLD H-NAFLD_P_ Value L-NAFLD Versus Control H-NAFLD Versus Control H-NAFLD Versus L-NAFLD Women ( n = 109)N 51 47 11 Age (years; mean ± SD) 57.4 ± 6.1 55.4 ± 5.0 55.5 ± 5.5 .1 .36 .97 BMI (kg/m 2 ; mean ± SD) 29.0 ± 4.5 30.3 ± 3.6 30.1 ± 2.1 .13 .45 .86 T2* (ms; mean ± SD) 23.6 ± 3.2 20.6 ± 4.2 20.1 ± 3.8 <.001 ∗ .003 ∗ .72 FBS (mg/dL; mean ± SD) 99.9 ± 35.7 111.7 ± 36.3 127.8 ± 40.6 .12 .04 ∗ .24 HbA1c (%; mean ± SD) 0.7 ± 0.3 2.6 ± 0.5 2.7 ± 0.9 .004 ∗ .01 ∗ .90 LDL (mg/dL; mean ± SD) 122.0 ± 35.1 123.2 ± 44.4 108.8 ± 52.8 .88 .35 .39 HDL (mg/dL; mean ± SD) 67.8 ± 18.3 59.1 ± 14.5 60.2 ± 14.2 .01 ∗ .24 .83 TG (mg/dL; mean ± SD) 128.4 ± 70.1 174.8 ± 96.7 217.3 ± 84.5 .01 ∗ .01 ∗ .31 Hypertension (Y/N) 12/39 17/30 4/7 .17 .45 .99 Men ( n = 98)N 46 38 14 Age (years; mean ± SD) 58.3 ± 6.8 58.3 ± 5.8 54.2 ± 3.5 .97 .03 ∗ .02 ∗ BMI (kg/m 2 ; mean ± SD) 26.0 ± 2.8 29.0 ± 3.0 29.8 ± 3.2 <.001 ∗ <.001 ∗ .41 T2* (ms; mean ± SD) 22.2 ± 3.6 20.3 ± 4.4 17.7 ± 3.2 .03 ∗ <.001 ∗ .04 ∗ FBS (mg/dL; mean ± SD) 105.8 ± 39.7 118.7 ± 39.3 106.3 ± 21.0 .15 .97 .28 HbA1c (%; mean ± SD) 0.8 ± 0.3 1.7 ± 0.5 1.9 ± 0.8 .12 .15 .85 LDL (mg/dL; mean ± SD) 129.3 ± 34.0 114.9 ± 38.3 128.0 ± 48.8 .08 .91 .32 HDL (mg/dL; mean ± SD) 54.1 ± 11.8 50.2 ± 11.9 49.1 ± 10.9 .14 .18 .77 TG (mg/dL; mean ± SD) 133.1 ± 52.3 201.2 ± 127.4 152.3 ± 48.6 .002 ∗ .24 .18 Hypertension (Y/N) 10/36 11/27 4/10 .41 .45 .58

BMI, body mass index; FBS, fasting blood sugar; HbA1c, glycosylated hemoglobin; HDL, high-density lipoprotein; H-NAFLD, high enzyme level NAFLD; LDL, low-density lipoprotein; L-NAFLD, low enzyme level NAFLD; NAFLD, nonalcoholic fatty liver disease; SD, standard deviation; TG, triglyceride.

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Multivariate Analysis

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Table 3

Multivariate Logistic Regression Analysis for Association of T2* Value and Other Risk Factors With Low and High Enzyme Level NAFLD in Men and Women

Variable OR for L-NAFLD (Control as Reference) OR for H-NAFLD (Control as Reference) OR for H-NAFLD (L-NAFLD as Reference) OR (95% CI)P Value OR (95% CI)P Value OR (95% CI)P Value Women T2* 0.79 (0.69–0.91) .001 ∗ 0.67 (0.51–0.87) .004 ∗ NA FBS NA 1.01 (0.99–1.02) .21 NA HbA1c 1.09 (0.91–1.37) .19 0.98 (0.56–1.74) .96 NA HDL 0.97 (0.93–0.99) .059 NA NA TG 1.003 (0.99–1.01) .49 1.001 (0.99–1.01) .81 NA Men T2* 0.91 (0.8–1.03) .16 0.73 (0.58–0.91) .006 ∗ 0.74 (0.58–0.95) .01 ∗ Age NA 0.86 (0.7–1.04) .13 0.78 (0.64–0.96) .01 ∗ BMI 1.51 (1.2–1.9) <.001 ∗ 1.54 (1.13–2.09) .005 ∗ NA LDL 0.98 (0.96–1.00) .06 NA NA TG 1.006 (0.99–1.01) .17 NA NA

BMI, body mass index; CI, confidence interval; FBS, fasting blood sugar; HbA1c, glycosylated hemoglobin; HDL, high-density lipoprotein; H-NAFLD, high enzyme level NAFLD; LDL, low-density lipoprotein; L-NAFLD, low enzyme level NAFLD; NA, not applicable (variables with P ≥ .1 in univariate analysis); NAFLD, nonalcoholic fatty liver disease; OR, odds ratio; SD, standard deviation; TG, triglyceride.

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Discussion

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