Nonalcoholic steatohepatitis: association of insulin resistance and mitochondrial abnormalities.
OBJECTIVE
The pathogenesis of nonalcoholic steatohepatitis (NASH) is unknown. We tested the hypothesis that NASH is associated with 2 defects: (1) peripheral insulin resistance, which increases lipolysis, delivery of free fatty acids (FFA) to the liver, and hepatic fatty acid beta oxidation, thereby creating oxidative stress; and (2) an abnormality within the hepatocytes that might render them more susceptible to injury from oxidative stress.


METHODS
The hypothesis was tested by evaluation of (1) insulin resistance by a 2-step hyperinsulinemic (10 and 40 mU. m(-2). min(-1)) euglycemic clamp; (2) insulin effects on lipolysis by enrichment of [U-(13)C]glycerol; (3) frequency and severity of structural defects in hepatocyte mitochondria in vivo; (4) fatty acid beta oxidation from serum [beta-OH butyrate], release of water-soluble radioactivity from (3)H-palmitate by cultured fibroblasts and urinary dicarboxylic acid excretion; and (5) hepatic lipid peroxidation by immunohistochemical staining for 3-nitrotyrosine (3-NT). Subjects with NASH (n = 6-10 for different studies) were compared with those with fatty liver (n = 6) or normal controls (n = 6).


RESULTS
NASH and fatty liver were both associated with insulin resistance, with mean glucose infusion rates (normal/fatty liver/NASH) of step 1, 4.5/1.6/0.9; step 2, 9.5/7.7/4.5 (P < 0.03 for both steps). Although baseline rates of glycerol appearance were higher in those with NASH than in those with fatty liver (means, 14.6 vs. 21.6 micromol. kg(-1). min(-1); P < 0.05), neither group significantly suppressed glycerol appearance at insulin infusion rates of 10 mU. m(-2). min(-1). NASH was associated with loss of mitochondrial cristae and paracrystalline inclusions in 9 of 10 subjects, compared with 0 of 6 subjects with fatty liver. However, no evidence of a generalized defect in fatty acid beta oxidation was noted in any group. Also, mean [beta-OH butyrate] was highest in those with NASH (means, 90 vs. 110 vs. 160 micromol/L; P < 0.04). Increased staining for 3-NT was present in fatty liver, and even greater staining was seen in NASH.


CONCLUSIONS
These data indicate that peripheral insulin resistance, increased fatty acid beta oxidation, and hepatic oxidative stress are present in both fatty liver and NASH, but NASH alone is associated with mitochondrial structural defects.
Division of Gastroenterology/Hepatology, Department of Internal Medicine, Virginia Commonwealth University-Medical College of Virginia, Richmond, Virginia 23298-0711, USA. ajsanyal@hsc.vcu.edu
A J
AJ
Sanyal
C
C
Campbell-Sargent
F
F
Mirshahi
W B
WB
Rizzo
M J
MJ
Contos
R K
RK
Sterling
V A
VA
Luketic
M L
ML
Shiffman
J N
JN
Clore
D000328:Adult; D000368:Aged; D001706:Biopsy; D002478:Cells, Cultured; D003998:Dicarboxylic Acids / Q000652:urine; D005230:Fatty Acids, Nonesterified / Q000097:blood; D005234:Fatty Liver / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D015309:Glucose Clamp Technique; D005990:Glycerol / Q000097:blood; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology; D006801:Humans; D006946:Hyperinsulinism / Q000378:metabolism / Q000473:pathology; D007328:Insulin / Q000097:blood; D007333:Insulin Resistance; D015227:Lipid Peroxidation; D008066:Lipolysis; D008297:Male; D008875:Middle Aged; D008930:Mitochondria, Liver / Q000378:metabolism / Q000473:pathology*; D010168:Palmitates / Q000494:pharmacology; D014316:Tritium
D003998:Dicarboxylic Acids; D005230:Fatty Acids, Nonesterified; D007328:Insulin; D010168:Palmitates; D014316:Tritium; D005990:Glycerol