Serum levels of advanced glycation end-products (AGEs) and the decoy soluble receptor for AGEs (sRAGE) can identify non-alcoholic fatty liver disease in age-, sex- and BMI-matched normo-glycemic adults.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is a serious health problem affecting ~25% of the global population. While NAFLD pathogenesis is still unclear, multiple NAFLD parameters, including reduced insulin sensitivity, impaired glucose metabolism and increased oxidative stress are hypothesised to foster the formation of advanced glycation end-products (AGEs). Given the link of AGEs with end organ damage, there is scope to examine the role of the AGE/RAGE axis activation in liver injury and NAFLD. METHODS: Age, sex and body mass index matched normo-glycemic NAFLD adults (n = 58) and healthy controls (n = 58) were enrolled in the study. AGEs were analysed by liquid chromatography-mass spectrometry (CML, CEL), fluorescence (pentosidine, AGE fluorescence), colorimetry (fructosamine) and ELISA (sRAGE). Their association with liver function, inflammation, fibrosis and stage of NAFLD was examined. RESULTS: Early and advanced glycation end-products, except Nε-carboxymethyl-L-lysine (CML), were 10-30% higher, sRAGE levels 1.7-fold lower, and glycation/sRAGE ratios 4-fold higher in the NAFLD cases compared to controls. While AGEs presented weak to moderate correlations with indices of liver function and damage (AST/ALT, HOMA-IR, TNF-α and TGF-ß1), including sRAGE to characterize the AGEs/sRAGE axis strengthened the associations observed. High glycation/sRAGE ratios were associated with 1.3 to 14-fold likelihood of lower AST/ALT ratios. The sum of AGEs/sRAGE ratios accurately distinguished between healthy controls and NAFLD patients (area under the curve of 0.85). Elevated AGEs/sRAGE (>7.8 mmol/pmol) was associated with a 12-fold likelihood of the presence of NAFLD. CONCLUSION: These findings strengthen the involvement of AGEs-RAGE axis in liver injury and the pathogenesis of NAFLD.

Serum levels of vascular endothelial growth factor in non-alcoholic fatty liver disease.

BACKGROUND: This study aimed to assess the significance of serum levels of vascular endothelial growth factor (VEGF) in non-alcoholic fatty liver disease (NAFLD). METHODS: Sixty-seven consecutive NAFLD patients and 47 healthy controls who visited our liver clinics between May 2008 and December 2010 were included. The NAFLD diagnosis required elevated alanine aminotransferase and/or gamma-glutamyl transpeptidase levels, evidence of hepatic steatosis on ultrasound and/or liver histology, and exclusion of other causes of liver injury. Serum VEGF levels were determined by an enzyme immunoassay. Liver biopsy was obtained in 34 NAFLD patients. Histological lesions were scored by a liver histopathologist. RESULTS: Serum VEGF levels tended to be lower in matched NAFLD patients than in healthy controls (296±146 vs. 365±186 pg/mL, P=0.092); levels in patients with non-alcoholic steatohepatitis (NASH) also tended to be lower than in those with simple fatty liver (FL) (279±149 vs. 359±190 pg/mL, P=0.095); while VEGF levels were significantly lower in NASH patients than in healthy controls (279±149 vs. 365±186 pg/mL, P=0.041). VEGF levels offered poor predictability for the differentiation between NAFLD patients and controls or between NASH and FL patients. However, patients with high VEGF levels (≥300 pg/mL) were significantly more likely to have FL, either in the total NAFLD population (67% vs. 35%, P=0.019) or in the 34 NAFLD patients with liver biopsy (57% vs. 15%, P=0.023), while those with high VEGF levels also had a significantly lower mean fibrosis score (0.7±0.9 vs. 1.6±1.0, P=0.017). CONCLUSION: Our data suggest that serum VEGF levels are equally high in healthy controls and in patients with simple fatty liver, but tend to decrease when NASH develops.

Increased abdominal fat levels measured by bioelectrical impedance are associated with histological lesions of nonalcoholic steatohepatitis.

BACKGROUND/AIM: Abdominal fat is considered to play an important role in nonalcoholic fatty liver disease (NAFLD), although it is not adequately studied because abdominal fat levels cannot be estimated easily. In this study, associations between abdominal obesity, as assessed by abdominal bioelectrical impedance analysis (BIA), and the characteristics of patients with NAFLD were explored. PATIENTS AND METHODS: Seventy-four consecutive NAFLD patients who underwent measurement of abdominal fat levels by BIA were included. Levels of abdominal fat 12.5 or less and more than 12.5 were considered to be average and increased, respectively. RESULTS: The mean±SD BMI was 30±4 kg/m and the mean abdominal fat levels were 16±5, whereas 26% of patients had average abdominal fat levels. Patients with average compared with those with increased abdominal fat levels were more frequently women (50 vs. 12%, P=0.001), had lower BMI (27±3 vs. 31±4 kg/m, P<0.001), lower Homeostasis Model Assessment index (2.6±1.4 vs. 3.9±2.7, P=0.045), and lower median liver stiffness on transient elastography (5.3 vs. 6.8 kPa, P=0.025). In patients with available liver biopsy, steatohepatitis was present more frequently in patients with increased compared with average abdominal fat levels (78 vs. 38%, P=0.030) and in patients with BMI 30 or more compared with less than 30 kg/m (87 vs. 48%, P=0.033), but similar in patients with increased or normal waist circumference (67 vs. 56%, P=0.693). CONCLUSION: Average levels of abdominal fat, as assessed by abdominal BIA, are mainly present in female patients with NAFLD and are associated with a lower degree of insulin resistance. Increased abdominal fat as assessed by BIA and obesity seem to represent strong risk factors for histological steatohepatitis.

Blood redox status is associated with the likelihood of nonalcoholic fatty liver disease irrespectively of diet's total antioxidant capacity.

It is well established that oxidative stress is implicated in nonalcoholic fatty liver disease pathogenesis, whereas the dietary intake of antioxidants has been reported to be low in patients with the disease. We hypothesized that blood redox status measurements would be associated with nonalcoholic fatty liver disease presence and severity, and that diet's total antioxidant capacity could moderate the aforementioned association. The study sample consisted of 73 patients with nonalcoholic fatty liver disease, of which 58 were matched by age, sex, and body mass index with 58 controls. Diet's total antioxidant capacity was estimated through the ferric-reducing antioxidant power, the total radical-trapping antioxidant parameter, and the Trolox equivalent antioxidant capacity scores, whereas blood redox status was assessed by measuring thiobarbituric acid reactive substances levels, the enzymatic activity of glutathione peroxidase, and serum resistance to oxidation. Diet's total antioxidant capacity scores and glutathione peroxidase activity were not significantly associated with the disease presence or severity. Both thiobarbituric acid reactive substances and serum resistance to oxidation were significantly associated with the likelihood of nonalcoholic fatty liver disease (odds ratios [ORs], 7.769 [P= .007] and 0.936 [P= .033], respectively), independently of abdominal fat level, degree of insulin resistance, blood lipid levels, markers of subclinical inflammation, and diet's total antioxidant capacity, but not with the disease histologic severity or stage. Our results support the association between blood redox status and the likelihood of nonalcoholic fatty liver disease regardless of diet's total antioxidant capacity.

The impact of cereal grain consumption on the development and severity of non-alcoholic fatty liver disease.

PURPOSE: There is evidence that dietary habits contribute to the presence and severity of non-alcoholic fatty liver disease (NAFLD). The aim of the present study was to explore any associations between consumption of grains and the development and severity of NAFLD. METHODS: Seventy-three consecutive NAFLD patients were enrolled. Additionally, 58 controls matched for age, sex and body mass index with 58 patients were also included. Consumption of grains was estimated through a semi-quantitative food frequency questionnaire. Medical history, anthropometric indices, body composition analysis, physical activity data, biochemical and inflammatory markers were available for all the participants. Liver stiffness measurement by transient elastography was performed in 58 and liver biopsy in 34 patients. RESULTS: In patients, consumption of whole grains was associated with lower abdominal fat level (ß = -0.24, p = 0.02) and lower levels of insulin resistance index (ß = -0.28, p = 0.009), while it also correlated inversely with interleukin-6 levels (ρ = -0.23, p = 0.05). Consumption of whole grains was associated with lower likelihood of having histological steatohepatitis (OR 0.97, 95% CI 0.94-1.000), after adjusting for sex and energy intake, but the association became weaker after further adjusting for abdominal fat or interleukin-6 levels. In the case-control analysis, consumption of refined grains was associated with higher odds of having NAFLD (OR 1.021, 95% CI 1.001-1.042), after adjusting for age, sex, energy intake, abdominal fat level, HOMA-IR, LDL, adiponectin and TNF-α. CONCLUSIONS: Although refined grain consumption increased the likelihood of having NAFLD, whole-grain consumption favorably affected clinical characteristics of patients with NAFLD and tended to be associated with less severe disease.

Adherence to the Mediterranean diet is associated with the severity of non-alcoholic fatty liver disease.

BACKGROUND & AIMS: Nutrition has been proposed as a potential environmental factor affecting the risk of non-alcoholic fatty liver disease (NAFLD). In the present study, the impact of adherence to the Mediterranean diet (MD) on the presence and severity of NAFLD was explored. METHODS: Seventy-three consecutive adult patients with recent NAFLD diagnosis were included. Adherence to the MD was estimated with MedDietScore. Demographic and anthropometric data, body composition analysis and several biochemical and inflammatory markers were estimated. Liver stiffness measurements by transient elastography were available in 58 patients and liver biopsies in 34 patients. Fifty-eight patients were matched with 58 healthy controls in terms of age, sex and body mass index. RESULTS: MedDietScore was negatively correlated to patients' serum alanine aminotransferase (p = 0.03) and insulin levels (p = 0.001), insulin resistance index (p = 0.005) and severity of steatosis (p = 0.006) and positively to serum adiponectin levels (p = 0.04). Patients with non-alcoholic steatohepatitis (NASH) exhibited lower adherence to MD (29.3 ± 3.2 vs. 34.1 ± 4.4, p = 0.004) compared to those with simple fatty liver. Logistic regression analysis revealed that one unit increase in the MedDietScore was associated with 36% lower likelihood of having NASH (odds ratio: 0.64, 95% confidence interval: 0.45-0.92), after adjusting for sex and abdominal fat level. No difference in the MedDietScore was observed between patients and controls. CONCLUSIONS: Higher adherence to the Mediterranean diet is not associated with lower likelihood of having NAFLD, but it is associated with less degree of insulin resistance and less severe liver disease among patients with NAFLD.

Changes of HBsAg and interferon-inducible protein 10 serum levels in naive HBeAg-negative chronic hepatitis B patients under 4-year entecavir therapy.

BACKGROUND & AIMS: Serum HBsAg levels might represent an important predictor of sustained off-treatment response in HBeAg-negative chronic hepatitis B (CHB). We evaluated the changes of HBsAg and interferon-inducible protein 10 (IP10) serum levels in HBeAg-negative CHB patients treated with entecavir. METHODS: 114 patients received entecavir for a median of 4.3 years. HBsAg levels were determined at baseline, 6 and 12 months and every year thereafter until year-4. IP10 levels were measured at baseline and annually until year-4 in 76 patients. RESULTS: Virological remission rates were high (year-1: 94%, after year-2: 97-98%). Compared to baseline, HBsAg levels decreased by a median of 0.03, 0.13, 0.17, 0.22, and 0.32 log10 IU/ml at 6 months and 1, 2, 3, and 4 years, respectively (p≤0.001 for all comparisons). The proportions of patients with HBsAg decline of ≥0.5 or ≥1 log10 IU/ml were 9% or 6% at year-1 and 21% or 10% at the last visit. Median IP10 levels (pg/ml) did not change from baseline to year-1 or -2 (245 vs. 229 or 251), but increased at year-3 and -4 (275 and 323, p<0.030). HBsAg drop ≥0.5 log10 was associated with baseline IP10 or IP10 >350 pg/ml (p≤0.002). HBsAg loss occurred in 4/114 (3.5%) patients or in 1/2, 3/21, and 0/91 patients with baseline HBsAg <100, 100-1000 and >1000 IU/ml, respectively (p<0.001). CONCLUSIONS: In HBeAg-negative CHB patients, 4-year entecavir therapy decreases serum HBsAg levels, but the rate of decline is rather slow. Serum IP10 levels represent a promising predictor of HBsAg decline in this setting.

The severity of histologic liver lesions is independent of body mass index in patients with nonalcoholic fatty liver disease.

GOALS/BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is associated with obesity, but it may also be present in lean individuals. We evaluated the characteristics of NAFLD patients, focusing on those with normal body mass index (BMI). STUDY: One hundred and sixty-two of 185 consecutive NAFLD patients were included. Demographic, clinical, somatometric, and laboratory characteristics were recorded. BMI<25 kg/m2 was considered to be normal. RESULTS: Of the 162 patients, 12% had normal BMI. Patients with normal compared with those with increased BMI fulfilled more frequently no criterion of metabolic syndrome (43% vs. 2%; P<0.001) and had higher median alanine aminotransferase (92 vs. 62 IU/L; P=0.032) and aspartate aminotransferase levels (45 vs. 37 IU/L; P=0.036). Liver stiffness values by transient elastography were significantly lower in patients with normal than in those with increased BMI (5.0 ± 1.6 vs. 9.5 ± 8.7 kPa; P=0.003). In the 56 patients with liver biopsy, the prevalence of nonalcoholic steatohepatitis (50% vs. 68.8%; P=0.423) and the severity of inflammation and fibrosis did not significantly differ between cases with normal and those with increased BMI. CONCLUSIONS: Approximately 1 of 8 NAFLD patients coming to a Greek tertiary liver center has normal BMI. On liver biopsy, normal BMI patients often have nonalcoholic steatohepatitis and histologic liver lesions of similar severity to the overweight or obese patients.