Obese Nonalcoholic Fatty Liver Disease Patients Research Articles

Structurally-engineered fatty acid 1024 (SEFA-1024) improves diet-induced obesity, insulin resistance, and fatty liver disease.

Obesity is a global epidemic that drives morbidity and mortality through cardiovascular disease, diabetes, and non-alcoholic fatty liver disease (NAFLD). No definitive therapy has been approved to improve glycemic control and treat NAFLD in obese patients. Here, we investigated a semi-synthetic, long chain, structurally-engineered fatty acid-1024 (SEFA-1024), as a treatment for obesity-induced hyperglycemia, insulin-resistance, and fatty liver disease in rodent models. A single dose of SEFA-1024 was administered to evaluate glucose tolerance and active glucagon-like peptide 1 (GLP-1) in lean rats in the presence and absence of a DPP-4 inhibitor. The effects of SEFA-1024 on weight loss and glycemic control were assessed in genetic (ob/ob) and environmental (high-fat diet) murine models of obesity. Liver histology, serum liver enzymes, liver lipidomics, and hepatic gene expression were also assessed in the high-fat diet murine model. SEFA-1024 reversed obesity-associated insulin resistance and improved glycemic control. SEFA-1024 increased active GLP-1. In a long-term model of diet-induced obesity, SEFA-1024 reversed excessive weight gain, hepatic steatosis, elevated liver enzymes, hepatic lipotoxicity, and promoted fatty acid metabolism. SEFA-1024 is an enterohepatic-targeted, eicosapentaenoic acid derivative that reverses obesity-induced dysregulated glucose metabolism and hepatic lipotoxicity in genetic and dietary rodent models of obesity. The mechanism by which SEFA-1024 works may include increasing aGLP-1, promoting fatty acid oxidation, and inhibiting hepatic triglyceride formation. SEFA-1024 may serve as a potential treatment for obesity-related diabetes and NAFLD.

Association between the LRP5 rs556442 gene polymorphism and the risks of NAFLD and CHD in a Chinese Han population

BackgroundMultiple studies have demonstrated the involvement of low-density lipoprotein receptor-related protein 5 (LRP5) in metabolism-related diseases. This study explored the relationship between the LRP5 rs556442 gene polymorphism and the risks of non-alcoholic fatty liver disease (NAFLD) and coronary heart disease (CHD) in a Chinese Han population.MethodsThis retrospective case–control study included 247 patients with NAFLD, 200 patients with CHD, 118 patients with both NAFLD and CHD, and 339 healthy controls from June 2018 to June 2019 at Qingdao Municipal Hospital. Basic information and clinical characteristics were collected for all subjects. The genotype and allele frequency of LRP5 rs556442 were determined.ResultsThe genotype distributions of LRP5 rs556442 differed significantly between the CHD and NAFLD + CHD groups (P < 0.05). The LRP5 rs556442 GG genotype markedly promoted the risk of NAFLD in CHD patients [odds ratio (OR) = 2.857, 95% confidence interval (CI): 1.196–6.824, P = 0.018). After adjustment for sex, age, and body mass index (BMI), this association remained significant (OR = 3.252, 95% CI: 1.306–8.102, P = 0.011). In addition, the LRP5 rs556442 AA + AG genotype was associated with an increased BMI in obese NAFLD patients (OR = 1.526, 95% CI: 1.004–2.319, P = 0.048). However, after adjustment for sex and age, this association was no longer significant (OR = 1.504, 95% CI: 0.991–2.282, P = 0.055).ConclusionsThis study found that the LRP5 rs556442 GG genotype increased the risk of NAFLD in CHD patients and AA + AG genotype may be associated with an increased BMI in obese NAFLD patients among a Chinese Han population.Trial registration ChiCTR, ChiCTR1800015426. Registered 28 March 2018—Retrospectively registered, http://www.chictr.org.cn/showproj.aspx?proj=26239.

Influence of endotoxemy on development of non-alcoholic fatty liver disease

Objective — to investigate endotoxin levels and to assess the relationship between endotoxemia and metabolic parameters and the degree of steatosis and fibrosis in patients with non‑alcoholic fatty liver disease (NAFLD) against the background of metabolic syndrome (MS).&#x0D; Materials and methods. The study included 68 patients with NAFLD with metabolic disorders and 20 people of the control group. Women prevailed in both investigational groups, the mean age was 49.34 ± 10.8 years. All patients underwent evaluation of their anthropometric parameters, investigation of body composition with determination of the percentage of visceral adipose tissue (VAT). To reveal VAT dysfunction, visceral adiposity index (VAI) was calculated with M. C. Amato method. Steatometry was performed by determining the wave attenuation coefficient and carrying out shear wave elastography. Endotoxin levels in blood serum were determined using a set of reagents «LAL Chromogenic Endpoint Assay» manufactured by «Hycult Biotech» (Netherlands).&#x0D; Results. Bacterial endotoxin levels in peripheral blood in the examined patients were almost three times higher in comparison with the control group (p &lt; 0.05). Patients with overweight and obesity had significantly higher endotoxin levels compared with the control group (p &lt; 0.05). The significant increase in endotoxin levels has been defined in obese NAFLD patients compared to the NAFLD patients with normal weight (p &lt; 0.05). The direct correlation was found between endotoxin parameters and parameters of abdominal and visceral obesity. In the group of patients with high VAT activity, the maximal values of this indicator were observed, which significantly differed from the corresponding indicator in patients with low and medium VAI (p &lt; 0.05). There was a direct correlation between moderate endotoxin and insulin resistance, a direct weak relationship between endotoxin and lipid profile parameters. In patients with grade 2 and 3 steatosis, significant differences in the level of endotoxemia were observed compared with the control group (p &lt; 0.05) and patients with grade 1 steatosis (p &lt; 0.05). At high grades of liver fibrosis (F3F4) there was a tendency to reduce endotoxemia compared with the low‑grade fibrosis (p &lt; 0.05).&#x0D; Conclusions. The revealed relationships between endotoxemia level and metabolic factors of NAFLD development indicate the pathogenetic role of bacterial endotoxemia in triggering a cascade of metabolic reactions with the subsequent accumulation of lipids in the liver tissue and the progression of steatosis and fibrosis.&#x0D;

Estimation of Liver Fat by FibroScan in Patients With Nonalcoholic Fatty Liver Disease.

BackgroundAssessment of nonalcoholic fatty liver disease (NAFLD) includes estimation of liver fat (steatosis). Controlled attenuation parameter (CAP) value obtained by FibroScan® (Echosens, Paris, France) is an alternative to liver biopsy for diagnosing and estimating steatosis (S). This study aimed to estimate the liver fat by CAP in NAFLD patients.MethodsAn observational cross-sectional study was conducted at the Liver Unit of Bir Hospital, from January 2021 to May 2021 after ethical clearance from the Institutional Review Board of the National Academy of Medical Sciences. A convenient sampling method was used. Data were analyzed with descriptive and inferential statistics involving bivariate and multivariate analysis.ResultsA total of 127 NAFLD patients were enrolled. The mean (±SD) CAP value was 271.53 (±50.69) dB/m. Total cholesterol, triglyceride, and body mass index (BMI) correlated positively (p<0.05) while systolic blood pressure correlated negatively with CAP value (p=0.031). On multivariate analysis, patients with BMI ≥25 kg/m2 were found 3.7 times more likely to have CAP ≥291 dB/m (S3, severe steatosis) than those with BMI <25 kg/m2 (p=0.048, 95% CI 1.01, 13.50). The mean (±SD) CAP values were 276.19 (±49.93) and 246.60 (±48.50) dB/m among those with BMI ≥25 kg/m2 and <25 kg/m2, respectively (p=0.016, using independent t-test). CAP steatosis grading correlated positively with both the ultrasound grading (p<0.001) and fibrosis grading by liver stiffness measurement (p=0.004). ConclusionIn this observational cross-sectional study of NAFLD patients, the mean (±SD) CAP value was 271.53 (±50.69) dB/m, which corresponds to moderate steatosis (S2). Obese NAFLD patients with ≥25 kg/m2 were 3.7 times more likely to have severe steatosis (S3) than nonobese NAFLD patients with BMI <25 kg/m2.

Performance of non-invasive fibrosis scores in non-alcoholic fatty liver disease with and without morbid obesity

BackgroundNon-invasive scores, such as the non-alcoholic fatty liver disease (NAFLD) Fibrosis Score (NFS), are increasingly used for liver fibrosis assessment in patients with NAFLD. The aim of this study was to assess the applicability and reliability of non-invasive fibrosis scores in NAFLD patients with and without morbid obesity.MethodsThree hundred sixty-eight patients with biopsy-proven NAFLD identified between January 2012 and December 2015 were studied; 225 with morbid obesity (biopsy obtained during bariatric surgery) and 143 patients without (termed as “conventional”).ResultsMedian age was 47 years, 57% were female. Median body mass index (BMI) was 42.9 kg/m2 with significant differences between our conventional and morbidly obese patients (BMI 29.0 vs. 50.8 kg/m2, p < 0.001). Overall, 42% displayed mild/moderate and 16% advanced liver fibrosis (stage III/IV). All tested scores were significantly linked to fibrosis stage (p < 0.001 for all). FIB-4 (AUROC 0.904), APRI (AUROC 0.848), and NFS (AUROC 0.750) were identified as potent predictors of advanced fibrosis, although NFS overestimated fibrosis stage in morbid obesity. Limiting BMI to a maximum of 40 kg/m2 improved NFS’ overall performance (AUROC 0.838). FIB-4 > 1.0 indicated high probability of advanced fibrosis (OR = 29.1). FIB-4 predicted advanced fibrosis independently from age, sex, BMI, and presence of morbid obesity.ConclusionsOur data suggest that FIB-4 score is an accurate predictor of advanced fibrosis in NAFLD throughout all BMI stages. Without adjustment, NFS tends to overestimate fibrosis in morbidly obese NAFLD patients. This problem may be solved by implementation of an upper BMI limit (for NFS) or adjustment of diagnostic thresholds.

Adiponectin, Leptin, and IGF-1 Are Useful Diagnostic and Stratification Biomarkers of NAFLD.

Background: Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease where liver biopsy remains the gold standard for diagnosis. Here we aimed to evaluate the role of circulating adiponectin, leptin, and insulin-like growth factor 1 (IGF-1) levels as non-invasive NAFLD biomarkers and assess their correlation with the metabolome.Materials and Methods: Leptin, adiponectin, and IGF-1 serum levels were measured by ELISA in two independent cohorts of biopsy-proven obese NAFLD patients and healthy-liver controls (discovery: 38 NAFLD, 13 controls; validation: 194 NAFLD, 31 controls) and correlated with clinical data, histology, genetic parameters, and serum metabolomics.Results: In both cohorts, leptin increased in NAFLD vs. controls (discovery: AUROC 0.88; validation: AUROC 0.83; p < 0.0001). The leptin levels were similar between obese and non-obese healthy controls, suggesting that obesity is not a confounding factor. In the discovery cohort, adiponectin was lower in non-alcoholic steatohepatitis (NASH) vs. non-alcoholic fatty liver (AUROC 0.87; p < 0.0001). For the validation cohort, significance was attained for homozygous for PNPLA3 allele c.444C (AUROC 0.63; p < 0.05). Combining adiponectin with specific serum lipids improved the assay performance (AUROC 0.80; p < 0.0001). For the validation cohort, IGF-1 was lower with advanced fibrosis (AUROC 0.67, p < 0.05), but combination with international normalized ratio (INR) and ferritin increased the assay performance (AUROC 0.81; p < 0.01).Conclusion: Serum leptin discriminates NAFLD, and adiponectin combined with specific lipids stratifies NASH. IGF-1, INR, and ferritin distinguish advanced fibrosis.

Low Skeletal Muscle Mass Is a Risk Factor for Subclinical Atherosclerosis in Patients with Nonalcoholic Fatty Liver Disease.

Sarcopenia and nonalcoholic fatty liver disease (NAFLD) are associated with subclinical atherosclerosis. We aimed to investigate the association between low skeletal muscle mass (LSMM) and subclinical atherosclerosis in patients with NAFLD. A total of 683 patients with ultrasound-confirmed NAFLD who underwent carotid ultrasonography were enrolled retrospectively. The appendicular skeletal muscle mass divided by the body mass index was used to define LSMM. Using carotid ultrasound, increased carotid intima–media thickness (cIMT, >1 mm) and the presence of carotid plaques were measured. Of the 683 patients, 75 (11.0%) had LSMM. In multivariate analyses, LSMM was associated with increased cIMT (odds ratios (ORs) = 2.26 to 2.95, all p < 0.05) and carotid plaques (ORs = 2.05 to 2.90, all p < 0.05). The proportion of increased cIMT and carotid plaques was significantly higher in obese NAFLD patients with LSMM than in those without LSMM (33.3% vs. 17.6% for cIMT and 12.7% vs. 5.7% for carotid plaques, respectively; p < 0.001). Furthermore, LSMM was independently associated with increased cIMT (ORs = 2.44 to 3.30, all p < 0.05) and carotid plaques (ORs = 2.56 to 3.54, all p < 0.05) in obese NAFLD patients. LSMM is associated with subclinical atherosclerosis in patients with NAFLD.

Variation in Lifestyle-Related Behavior Among Obese Indian Patients With Non-alcoholic Fatty Liver Disease

Lifestyle modification is the mainstay of treatment in Non-Alcoholic Fatty Liver Disease (NAFLD). Published Indian data on the diet and lifestyle of Indian NAFLD patients is scarce. This study explored variation in lifestyle-related behavior among obese patients with NAFLD. Ultrasonography (USG) diagnosed obese NAFLD patients (n = 140) were assessed for dietary intake [1-day 24 hours recall, food-frequency questionnaire (FFQ)] and physical activity (PA) [Global Physical Activity Questionnaire (GPAQ)]. Diet quality score (0–30) and physical activity (PA) levels were used to study variation in lifestyle and assess the effect of lifestyle on the severity of NAFLD. Compared to the recommendation, calorie consumption was 25.2% higher in nearly half (53.6%) of the subjects and mean macronutrient intake was imbalanced (60.3% carbohydrates, 12.4% protein, 25.5% fats). Variation was seen in terms of diet quality—good (3%), moderate (54.3%), or poor (43.5%) and intensity of PA—high (15%), moderate (47.9%), or low (37.1%). No patient had a combination of high PA and good diet quality within all grades of NAFLD. Our study suggests wide variation in lifestyles of obese patients with NAFLD and need for a more flexible and individualized modification of their diet and PA.

Low serum vitamin D concentrations are associated with obese but not lean NAFLD: a cross-sectional study

BackgroundA low serum vitamin D concentration has been reported to be associated with an increased risk of non-alcoholic fatty liver disease (NAFLD); however, whether lean or obese individuals show a similar association between vitamin D and NAFLD remains speculative. This study aimed to explore the relationship between serum vitamin D concentration and NAFLD in lean and obese Chinese adults.MethodsThis cross-sectional study included 2538 participants (1360 men and 1178 women) who underwent health checkups at the First Affiliated Hospital, Zhejiang University School of Medicine in 2019. NAFLD was diagnosed by liver ultrasound excluding other causes. The association of serum vitamin D concentration with NAFLD was analyzed in lean and obese participants.ResultsThe overall prevalence of NAFLD was 33.61% (13.10% in lean and 53.32% in obese) in this study population. The serum vitamin D levels of obese NAFLD patients were lower than those of obese NAFLD-free controls. However, the serum vitamin D levels of lean NAFLD patients were comparable to those of lean NAFLD-free controls. Serum vitamin D level was negatively correlated with the prevalence of NAFLD in obese but not lean participants. Serum vitamin D level was independently associated with the risk of NAFLD in obese participants, with an adjusted odds ratio (95% CI) of 0.987 (0.981–0.993). However, serum vitamin D level was not related to the risk of NAFLD in lean participants.ConclusionsA low serum vitamin D level is associated with NAFLD in obese but not lean participants.

N6 -Methyladenosine Reader Protein YT521-B Homology Domain-Containing 2 Suppresses Liver Steatosis by Regulation of mRNA Stability of Lipogenic Genes.

Nonalcoholic fatty liver disease (NAFLD) is characterized by accumulation of excessive triglycerides (TGs) in hepatocytes. Obesity is a major risk factor for developing fatty liver, although the intracellular molecular basis remains largely unclear. N6 -methyladenosine (m6 A) RNA methylation is the most common internal modification in eukaryotic mRNA. In the present study, by m6 A sequencing and RNA sequencing, we found that both m6 A enrichment and mRNA expression of lipogenic genes were significantly increased in leptin-receptor-deficient db/db mice. Importantly, our results showed that YT521-B homology domain-containing 2 (Ythdc2), an m6 A reader, was markedly down-regulated in livers of obese mice and NAFLD patients. Suppression of Ythdc2 in livers of lean mice led to TG accumulation, whereas ectopic overexpression of Ythdc2 in livers of obese mice improved liver steatosis and insulin resistance. Mechanistically, we found that Ythdc2 could bind to mRNA of lipogenic genes, including sterol regulatory element-binding protein 1c, fatty acid synthase, stearoyl-CoA desaturase 1, and acetyl-CoA carboxylase 1, to decrease their mRNA stability and inhibit gene expression. Our findings describe an important role of the m6 A reader, Ythdc2, for regulation of hepatic lipogenesis and TG homeostasis, which might provide a potential target for treating obesity-related NAFLD.

Lipoprotein and Metabolic Profiles Indicate Similar Cardiovascular Risk of Liver Steatosis and NASH

Background and Aim: Nonalcoholic fatty liver disease (NAFLD) affects about 25% of the global population, with no reliable noninvasive tests to diagnose nonalcoholic steatohepatitis (NASH) and to differentiate between NASH and nonalcoholic fatty liver (NAFL) (steatosis alone). It is unclear if NAFL and NASH differ in cardiovascular risk for patients. Here, we compared obese NAFLD patients with a healthy cohort to test whether cholesterol compounds could represent potential noninvasive markers and to estimate associated risks. Method: Serum samples of 46 patients with histologically confirmed NAFLD (17 NAFL, 29 NASH) who underwent bariatric surgery were compared to 32 (9 males, 21 females) healthy controls (HCs). We analyzed epidemiological data, liver enzymes, cholesterol and lipid profile, and amino acids. The latter were analyzed by nuclear magnetic resonance spectroscopy. Results: Total serum and high-density lipoprotein (HDL) cholesterol were significantly lower in the NAFLD group than in HCs, with a stronger reduction in NASH. Similar observations were made for sub-specification of HDL-p, HDL-s, SHDL-p, and LHDL-p cholesterols. Low-density lipoprotein (LDL)-s and LLDL-p cholesterol were significantly reduced in NAFLD groups. Interestingly, SLDL-p cholesterol was significantly higher in the NAFL group with a stronger elevation in NASH than in HCs. The amino acids alanine, leucin, and isoleucine were significantly higher in the NAFL and NASH groups than in HCs. Conclusion: We show in this study that cholesterol profiles, apolipoproteins, and amino acids could function as a potential noninvasive test to screen for NAFLD or even NASH in larger populations. However, few differences in cholesterol profiles were identified between the NAFL and NASH groups, indicating similar cardiovascular risk profiles.

The Nuclear Orphan Receptor NR2F6 Promotes Hepatic Steatosis through Upregulation of Fatty Acid Transporter CD36.

Nuclear receptors (NRs) are a superfamily of transcription factors which sense hormonal signals or nutrients to regulate various biological events, including development, reproduction, and metabolism. Here, this study identifies nuclear receptor subfamily 2, group F, member 6 (NR2F6), as an important regulator of hepatic triglyceride (TG) homeostasis and causal factor in the development of non‐alcoholic fatty liver disease (NAFLD). Adeno‐associated virus (AAV)‐mediated overexpression of NR2F6 in the liver promotes TG accumulation in lean mice, while hepatic‐specific suppression of NR2F6 improves obesity‐associated hepatosteatosis, insulin resistance, and methionine and choline‐deficient (MCD) diet‐induced non‐alcoholic steatohepatitis (NASH). Mechanistically, the fatty acid translocase CD36 is identified as a transcriptional target of NR2F6 to mediate its steatotic role. NR2F6 is able to bind directly onto the CD36 promoter region in hepatocytes and increases the enrichment of nuclear receptor coactivator 1 (SRC‐1) and histone acetylation at its promoter. Of pathophysiological significance, NR2F6 is significantly upregulated in the livers of obese mice and NAFLD patients. Moreover, treatment with metformin decreases NR2F6 expression in obese mice, resulting in suppression of CD36 and reduced hepatic TG contents. Therefore, these results provide evidence for an unpredicted role of NR2F6 that contributes to liver steatosis and suggest that NR2F6 antagonists may present a therapeutic strategy for reversing or treating NAFLD/NASH pathogenesis.

Circulating lipidomic alterations in obese and non-obese subjects with non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) affects obese and non-obese individuals. However, mechanisms underlying non-obese non-alcoholic steatohepatitis (NASH) remain unclear. To attempt to identify metabolic perturbations associated with non-obese and obese NAFLD using a lipidomics approach. A cross-sectional analysis of 361 subjects with biopsy-proven NAFLD (157 NAFL and 138 NASH) and healthy controls (n=66) was performed. Individuals were categorised as obese or non-obese based on the Asian cut-off for body mass index. Circulating lipidomic profiling of sera was performed based on the histological severity of NAFLD. Circulating lipidomic alterations were validated with an independent validation set (154 NAFLD subjects [93 NAFL and 61 NASH] and 21 healthy controls). Saturated sphingomyelin (SM) species were significantly associated with visceral adiposity in non-obese NAFLD (SM d38:0; P<0.001) but not in obese NAFLD. Additionally, SM levels were significantly associated with systemic and adipose tissue insulin resistance (SM d38:0; P=0.002 and <0.001, respectively). Five potential lipid metabolites for non-obese subjects and seven potential lipids for obese subjects were selected to predict NAFLD and NASH. These lipid combinations showed good diagnostic performance for non-obese (area under the curve [AUC] for NAFLD/NASH=0.916/0.813) and obese (AUC for NAFLD/NASH=0.967/0.812) subjects. Moreover, distinctly altered patterns of diacylglycerol (DAG), triacylglycerol (TAG) and SM levels were confirmed in the validation set depending on the histological severity of NAFLD. Non-obese and obese NAFLD subjects exhibit unique circulating lipidomic signatures, including DAGs, TAGs and SMs. These lipid combinations may be useful biomarkers for non-obese and obese NAFLD patients.

Coping strategies predicting quality of life in diabetic and obese NAFLD patients

Abstract Background To date, coping strategies have not been studied in patients with nonalcoholic fatty liver disease (NAFLD), despite evidence of their relevance in chronic liver pathology, Type 2 diabetes mellitus (T2DM) and obesity (OB). We therefore analyzed which coping strategies predicted quality of life in diabetic and obese NAFLD patients. Methods Four hundred and ninety-two biopsy-proven NAFLD patients (290 men and 202 women, mean age 54.90±11.74) were evaluated using The Brief COPE, 12-Item Short-Form Health Survey (SF-12) and Chronic Liver Disease Questionnaire-Non-Alcoholic Fatty Liver Disease (CLDQ-NAFLD). A stepwise multiple linear regression analysis was performed on four groups (G1, n = 335, absence of T2DM; G2, n = 157, presence of T2DM; G3, n = 249, absence of OB; and G4, n = 243, presence of OB) to analyze which coping strategies predicted patient quality of life (physical component summary SF-12, mental component summary SF-12, and total CLDQ-NAFLD). Results In both diabetic and obese patients, active coping (T2DM, p = 0.003, β = 0.26; OB, p = 0.000, β = 0.33) and denial (T2DM, p = 0.027, β=-0.19; OB, p = 0.004, β=-0.18) predicted the physical component summary. Denial (T2DM, p = 0.000, β=-0.30; OB, p = 0.001, β=-0.19), positive reframing (T2DM, p = 0.000, β = 0.28; OB, p = 0.000, β = 0.29), self-blame (T2DM, p = 0.000, β=-0.24; OB, p = 0.000, β=-0.26) and self-distraction (T2DM, p = 0.033, β=-0.13; OB, p = 0.023, β=-0.11) predicted the mental component summary. Denial (T2DM, p = 0.000, β=-0.34; OB, p = 0.000, β=-0.31), positive reframing (T2DM, p = 0.000, β = 0.30; OB, p = 0.005, β = 0.15) and self-blame (T2DM, p = 0.000, β=-0.26; OB, p = 0.000, β=-0.28) also predicted the total CLDQ-NAFLD in both groups. Conclusions Active coping and positive reframing predicted better quality of life, while denial, self-blame and self-distraction predicted worse quality of life in diabetic and obese NAFLD patients, suggesting the inclusion of coping strategies in future multidisciplinary NAFLD treatments. Key messages Importance of coping strategies for NAFLD patients: active coping and positive reframing predicted better quality of life, while denial, self-blame and self-distraction predicted worse quality. This study shows the need to design multidisciplinary strategies for managing NAFLD and improving patient quality of life, in which intervention in coping strategies should be a major element.

Effects of obesity reduction on transient elastography–based parameters in pediatric non-alcoholic fatty liver disease

AimTo clarify the effects of obesity reduction on non-alcoholic fatty liver disease (NAFLD) in obese children. MethodsTwenty-six obese pediatric NAFLD patients (median age, 13.0 years; range, 6.4–16.6 years), who underwent obesity management supported by regular hospital visits and/or hospital admission, were studied to explore how reductions in weight and body mass index (BMI) percentile affected two transient elastography–based parameters: controlled attenuation parameter (CAP) and liver stiffness (LS), which reflect the degree of hepatic fat deposition and liver fibrosis, respectively. ResultsUnivariate analysis revealed that CAP reduction was correlated positively with baseline CAP and reductions in weight and BMI percentile (r = 0.320–0.525), whereas LS reduction was correlated positively with baseline LS and reductions in weight, BMI percentile, aspartate transaminase, and alanine aminotransferase (r = 0.385–0.625). Multivariate analysis revealed that baseline CAP and reduction in weight were significantly associated with CAP reduction, whereas reduction in weight and alanine aminotransferase were significantly associated with LS reduction. Short-term weight control by hospital admission (24.9 ± 9.5 days) provided significantly higher reductions in weight and BMI percentile (both P < 0.001) and was associated with reductions of CAP and LS (P = 0.04 and 0.01) compared with regular hospital visit-supported self-directed weight management (0.9 ± 0.8 years). ConclusionsWeight reduction in obese pediatric NAFLD patients resulted in reduced hepatic fat deposition and liver stiffness. Weight control by short-term hospital admission is an alternative approach to regular hospital visit-supported self-directed weight management in NAFLD patients who fail to reduce obesity.

Non-obese patients with nonalcoholic fatty liver disease may use a lower liver stiffness cut-off to assess fibrosis stages.

We aimed to estimate the optimal cut-off values of liver stiffness measurement (LSM) for diagnosing and staging fibrosis in non-obese and obese patients with nonalcoholic fatty liver disease (NAFLD). NAFLD patients diagnosed by liver biopsy according to the Nonalcoholic Steatohepatitis Clinical Research Network scoring system were enrolled in this study. Non-obesity was defined as a body mass index (BMI) less than 25 kg/m2 . LSM was performed by experienced physicians within 2 weeks before or after liver biopsy. A total of 158 patients were included. Average BMI of the non-obese (n = 68) and obese (n = 90) groups was 23.2 ± 1.6 and 27.9 ± 2.5 kg/m2 , respectively. After adjusted for age, fibrosis stage, steatosis grade and type 2 diabetes mellitus, the obese group had a LSM of 3.522 kPa higher than the non-obese patients (P = 0.003). LSM values of the non-obese patients had a lower trend when stratified by fibrosis stage, especially in cirrhosis (F4; P = 0.021). Applying separate cut-off values for patients with NAFLD in individual fibrosis stage, 5.8 vs 7.5 kPa (≥ F1), 7.6 vs 8.5 kPa (≥ F2), 9.1 vs 11.2 kPa (≥ F3), and 12.5 vs 14.3 kPa (F4), improved their diagnostic odds ratios compared with overall cut-off values. In the non-obese NAFLD group, using a separate cut-off avoided underestimating 9.1% of patients with cirrhosis. Non-obese NAFLD group had lower LSM than the obese group. Different cut-off values should be used to measure liver fibrosis stage in non-obese and obese NAFLD patients.

Metagenome of Gut Microbiota of Children With Nonalcoholic Fatty Liver Disease.

Aim: To investigate the intestinal flora of nonalcoholic fatty liver disease (NAFLD) in Chinese children and adolescents using metagenomic approach.Methods: All participants underwent magnetic resonance spectroscopy (MRS) to quantify liver fat content. Hepatic steatosis was defined as MRS proton density fat fraction (MRS-PDFF) >5%. A total of 58 children and adolescents were enrolled in this study, including 25 obese NAFLD patients, 18 obese non-NAFLD children, and 15 healthy children. Stool samples were collected and analyzed with metagenomics. We used Shannon index to reflect the alpha diversities of gut microbiota. Wilcoxon rank sum test and Kruskal-Wallis test were performed to evaluate alpha diversities between groups. At last, the differences of gut microbiota composition and functional annotations between obese with and without NAFLD and healthy children were assessed by Kruskal-Wallis test.Results: Significant differences in gut microbiota composition and functional annotations among three groups of children and adolescents have been observed. Deep sequencing of gut microbiota revealed high abundance of phylum Proteobacteria (Gammaproteobacteria) in obese NAFLD patients, comparing with the control group. Overall, obese children without NAFLD had less abundant Helicobacter and Helicobacter pylori. Compared to the control group, in obese children with NAFLD, the abundance of Bacteroidetes (Alistipes) were significantly reduced. Faecalibacterium prausnitzii was the only species representing a difference between obese children with and without NAFLD. There were not significant differences in terms of alpha diversity among three groups. Functional annotations demonstrated that several pathways were differentially enriched between groups, including metabolism of other amino acids, replication and repair, folding, sorting, degradation, and glycan biosynthesis and metabolism.Conclusion: Significantly differences are observed in gut microbiota composition and functional annotations between obese children with and without NAFLD in comparison to the healthy children group. The characteristic of gut microbiota in this study may contribute to a further understanding the gut-liver axis of pediatric NAFLD in China.

PNPLA3 I148M Polymorphism in Patients with Nonalcoholic Fatty Liver Disease, Obesity and Prediabetes.

Nonalcoholic fatty liver disease (NAFLD) is closely associated with obesity and insulin resistance, and therefore predisposes to type 2 diabetes and cardiovascular diseases. Lipid deposition in the liver seems to be critical in the pathogenesis of NAFLD. A common genetic variant, the patatin-like phospholipase domain-containing protein 3 (PNPLA3) has been associated with NAFLD. The aim of the present study was to evaluate the association between PNPLA3, key gene of lipid metabolism and the metabolic traits in obesity NAFLD patients with and without prediabetes. A total of 208 obese NAFLD patients without (n=125) and with prediabetes (n=83) were included. The genotyping of PNPLA3 I148M variant (rs738409) was performed by restriction analysis. Regarding rs738409 (I148M) polymorphism, CG genotype was positively correlated with prediabetes, insulin resistance, dyslipidemia and metabolic syndrome compared to the wild CC genotype. The carriers of the PNPLA3 I148M variant have 9.6-fold higher risk of glucose disturbances compared to wild genotype (OR 9.649, 95%CI 2.100-44.328, р=0.004). The carriers of the PNPLA3 I148M variant also have a 3 times higher risk for the presence of metabolic syndrome (OR 2.939, 95% CI: 1.590-5.434, p=0.001) and a 2.1-fold higher risk for the presence of insulin resistance (OR 2.127, 95% CI: 1.078-4.194, p=0.029). PNPLA3 I148M is associated with increased risk of prediabetes, metabolic syndrome and insulin resistance in obese patients with NAFLD.

Circulating Hsp90 Isoform Levels in Overweight and Obese Children and the Relation to Nonalcoholic Fatty Liver Disease: Results from a Cross-Sectional Study.

Background Obesity prevalence is increasing in children. It is associated with various comorbidities including nonalcoholic fatty liver disease (NAFLD). Hsp90 isoforms were identified in previous proteomic studies as potential biomarkers for NAFLD. The aim of the study was to analyze circulating levels of Hsp90α and Hsp90β in overweight and obese children. In addition, Hsp90α and Hsp90β were evaluated as biomarkers for NAFLD in overweight and obese children. Methods 68 overweight and obese children and ten age- and gender-matched controls were recruited. Hsp90α and Hsp90β levels were analyzed from serum in both controls and overweight and obese children by ELISA. Results Serum Hsp90β and total Hsp90 levels were statistically significantly higher in overweight and obese children compared to controls. On the contrary, there was no difference in Hsp90α levels between overweight and obese children and healthy controls. Hsp90 isoforms had different expression in NAFLD patients. Hsp90β levels were higher in overweight and obese NAFLD patients while Hsp90α levels were lower. Hsp90α to Hsp90β ratio had better accuracy for NAFLD diagnosis in obese and overweight patients compared to individual biomarkers. Conclusion Hsp90 isoforms were confirmed on an independent cohort as biomarkers for NAFLD in overweight and obese children. In these patients, it seems to be more useful to separately analyze Hsp90 isoforms rather than total Hsp90 as the isoforms have greater discriminative capacity.

Early Predictors of Cardiovascular Disease Risk in Nonalcoholic Fatty Liver Disease: Non-obese Versus Obese Patients.

Nonalcoholic fatty liver disease (NAFLD) is regarded as a risk factor of cardiovascular disease (CVD). However, the association between non-obese NAFLD and CVD has not been well established. We aimed to compare the CVD risk between non-obese and obese NAFLD patients, and explored the factors associated with subclinical atherosclerosis. Consecutive NAFLD patients estimated by magnetic resonance imaging-based proton density fat fraction (MRI-PDFF) were recruited. Liver fat content (LFC) and liver stiffness were measured with MRI-PDFF and shear wave elastography, respectively. CVD risk was estimated by atherosclerosis index (AI), carotid intima-media thickness, carotid plaque, and Framingham risk score (FRS). This study included 543 NAFLD patients. The presence of carotid intima-media thickening and carotid plaque, FRS, and AI were comparable between non-obese and obese patients. Age increased per 10years (OR 9.68; P < 0.001) and liver fibrosis (liver stiffness > 6.1kPa, OR 4.42; P = 0.004) were significant factors associated with carotid intima-media thickening in non-obese patients, while age increased per 10years (OR 2.02; P < 0.001), liver fibrosis (OR 2.18; P = 0.039), and LFC > 10% (OR 2.29; P = 0.021) were independent predictors in obese patients. Only elevated triglyceride was significantly associated with carotid plaque in non-obese patients (OR 2.42; P = 0.033), while age increased per 10years (OR 1.77; P = 0.002) and LFC > 10% (OR 2.83; P = 0.019) were significant predictors in obese patients. Liver stiffness and age were strongly predictive of subclinical atherosclerosis in all NAFLD, while LFC was an additional predictor in obese NAFLD patients. Our findings highlight that early CVD screening strategy should be established for NAFLD patients according to different BMIs.