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Proteomic analysis of liver mitochondria from rats with nonalcoholic steatohepatitis

World Journal of Gastroenterology 2014年第16期20卷 4778-4786页

作者： Lin Li De-Zhao Lu You-Ming Li Xue-Qun Zhang Xin-Xin Zhou Xi Jin Department of Gastroenterology the First Affiliated HospitalCollege of MedicineZhejiang University College of Life Sciences Zhejiang Chinese Medical University

AIM:To explore mitochondrial dysfunction in nonalcoholic steatohepatitis(NASH)by analyzing the proteome of liver mitochondria from a NASH model.METHODS:The NASH rat model was established by feeding rats a fat-rich diet for 24 wk and was confirmed using hematoxylin and eosin staining of liver tissue and by changes in the levels of serum alanine transaminase,aspartate aminotransferase,triglyceride,total cholesterol and other markers.Liver mitochondria from each group were isolated using differential centrifugation.The mitochondrial samples were lyzed,purified and further analyzed using two-dimensional electrophoresis combined with matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry.Bioinformatic analyses of assigned gene ontology and biological pathway was used to study functional enrichments in the abundant proteomic data.RESULTS:Eight up-regulated and sixteen down-regulated proteins were identified that showed greater than1.5-fold differences between the controls and the NASH group.These dysregulated proteins were predicted to be involved in different metabolic processes including fatty acidβ-oxidation processes,lipid metabolic processes,cell-cycle arrest,cell polarity maintenance,and adenosine triphosphate/sex hormone metabolic processes.Novel proteins that may be involved in NASH pathogenesis including the trifunctional enzyme Hadha,thyroxine,prohibitin,aldehyde dehydrogenase ALDH1L2,UDP-glucuronosyltransferase 2B31,and carbamoyl-phosphate synthase were identified using bioinformatics tools.The decreased expression of Hadha in NASH liver was verified by Western blotting,which was used as a complementary technique to confirm the proteomic results.CONCLUSION:This novel report on the liver mitochondrial proteome of a NASH model may provide a reservoir of information on the pathogenesis and treatment of NASH.

关键词： nonalcoholic steatohepatitis Hadha Proteomics Rat

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Clinical differences between alcoholic liver disease and nonalcoholic fatty liver disease

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World Journal of Gastroenterology 2014年第26期20卷 8393-8406页

作者： Nobuyuki Toshikuni Mikihiro Tsutsumi Tomiyasu Arisawa Department of Gastroenterology Kanazawa Medical University Department of Hepatology Kanazawa Medical University

Alcoholic liver disease(ALD)and nonalcoholic fatty liver disease(NAFLD)are serious health problems worldwide.These two diseases have similar pathological spectra,ranging from simple hepatic steatosis to steatohepatitis,liver cirrhosis,and hepatocellular carcinoma.Although most subjects with excessive alcohol or food intake experience simple hepatic steatosis,a small percentage of individuals will develop progressive liver disease.Notably,both ALD and NAFLD are frequently accompanied by extrahepatic complications,including cardiovascular disease and malignancy.The survival of patients with ALD and NAFLD depends on various disease-associated conditions.This review delineates the clinical characteristics and outcomes of patients with ALD and NAFLD by comparing their epidemiology,the factors associated with disease susceptibility and progression,and the predictors and characteristics of outcomes.A comprehensive understanding of the characteristics and outcomes of ALD and NAFLD is imperative in the management of these chronic liver diseases.

关键词： Alcoholic liver disease nonalcoholic fatty liver d

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Role of liver biopsy in nonalcoholic fatty liver disease

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World Journal of Gastroenterology 2014年第27期20卷 9026-9037页

作者： ILKe Nalbantoglu Elizabeth M Brunt Department of Pathology and Immunology Washington University School of MedicineSaint Louis MO 63110 United States

nonalcoholic fatty liver disease(NAFLD), defined as abnormal accumulation(> 5%) of hepatic triglyceride without excess alcohol intake, is the most common form of chronic liver disease in adults and children in the United States. NAFLD encompasses a spectrum of histologic findings including uncomplicated steatosis, steatosis with inflammation and steatohepatitis [nonalcoholic steatohepatitis(NASH)]; the latter can advance to cirrhosis and hepatocellular carcinoma. NASH is currently accepted as the hepatic manifestation of the set of cardiovascular risk factors collectively known as metabolic syndrome. In 1999 a system for histologic grading and staging for NASH was proposed; this was revised by the NASH Clinical Research Network in 2005 for the entire spectrum of lesions in NAFLD, including the lesions and patterns of pediatric NAFLD, and for application in clinical research trials. Diagnosis remains distinct from grade and stage. A recent European proposal separates steatosis from activity to derive a numeric diagnosis of NASH. Even though there have been promising advancements in non-invasive testing, these tests are not yet detailed enough to replace the full range of findings provided by liver biopsy evaluation. Limitations of biopsy are acknowledged, but liver biopsy remains the "gold standard" for diagnosis and determination of amounts of necroinflammatory activity, and location of fibrosis, as well as remodeling of the parenchyma in NASH. This review focuses on the specific histologic lesions of NAFLD and NASH, grading and staging, differential diagnoses to be considered, and the continuing role of the liver biopsy in this important liver disease.

关键词： nonalcoholic fatty liver disease nonalcoholic stea

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Apolipoprotein C3(-455T>C) polymorphism confers susceptibility to nonalcoholic fatty liver disease in the Southern Han Chinese population

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World Journal of Gastroenterology 2014年第38期20卷 14010-14017页

作者： Min-Rui Li Sheng-hong Zhang Kang Chao Xian-hua Liao Jia-yan yao Min-hu Chen Bi-hui Zhong Division of Gastroenterology The First Affiliated HospitalSun Yat-sen University

AIM:To investigate the relationship between Apolipoprotein C3(APOC3)(-455T>C) polymorphism and nonalcoholic fatty liver disease(NAFLD) in the Southern Chinese han population.METHODS:In this prospective case-control study,we recruited 300 NAFLD patients and 300 healthy controls to a cohort representing Southern Chinese han population at The First Affiliated Hospital,Sun Yat-sen University,from January to December 2012. Polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing were used to genotype the APOC3(-455T>C) variants.RESULTS:After adjusting for age,gender,and bodymass index,TC and CC genotypes were found to increase the susceptibility to NAFLD compared to the TT genotype,with adjusted odds ratios(ORs) of 1.77(95%CI:1.16-2.72) and 2.80(95%CI:1.64-4.79),respectively. Further stratification analysis indicated that carriers of the CC genotype was more susceptible to insulin resistance(IR) than those of the TT genotype,with an OR of 3.24(95%CI:1.52-6.92). The CC genotype also was associated with a significantly higher risk of hypertension,hypertriglyceridemia,and low levels of high-density lipoprotein cholesterol(hDL)(P C) polymorphism and obesity,impaired glucose tolerance,hyperuricemia,hypercholesterolemia,or high levels of low-density lipoprotein cholesterol(LDL)(P > 0.05).CONCLUSION:APOC3(-455T>C) genetic variation is involved in the susceptibility to developing NAFLD,IR,hypertension,hypertriglyceridemia,and low hDL in the Southern Chinese han population.

关键词： Apolipoprotein C3 nonalcoholic fatty liver disease

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Linked PNPLA3 polymorphisms confer susceptibility to nonalcoholic steatohepatitis and decreased viral load in chronic hepatitis B

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World Journal of Gastroenterology 2015年第28期21卷 8605-8614页

作者： Qin Pan Rui-Nan Zhang Yu-Qin Wang Rui-Dan Zheng Yu-Qiang Mi Wen-Bin Liu Feng Shen Guang-Yu Chen Jia-Fa Lu Chan-Yan Zhu Shu-Yi Zhang Yi-Ming Chen Wan-Lu Sun Jian-Gao Fan Department of Gastroenterology Xinhua Hospital Shanghai Jiaotong University School of Medicine Diagnosis and Treatment Center for Liver Diseases Zhengxing HospitalZhangzhou Department of Infectious Diseases Tianjin Infectious Disease Hospital Wu-Jiao-Chang Community Health Center Clinical Epidemiology Center Shanghai Jiaotong University School of Medicine Shanghai Key Laboratory of Children’s Digestion and Nutrition

AIM:To investigate the association of PNPLA3 polymorphisms with concurrent chronic hepatitis B(CHB) and nonalcoholic fatty liver disease(NAFLD).METHODS:A cohort of Han patients with biopsyproven CHB,with or without NAFLD(CHB group,n = 51;CHB + NAFLD group,n = 57),and normal controls(normal group,n = 47) were recruited from Northern(Tianjin),Central(Shanghai),and Southern(Zhangzhou) China.Their PNPLA3 polymorphisms were genotyped by gene sequencing.The association between PNPLA3 polymorphisms and susceptibility to NAFLD,and clinical characteristics of NAFLD were evaluated on the basis of physical indices,liver function tests,glycolipid metabolism,and histopathologic scoring.The association of PNPLA3 polymorphisms and hepatitis B virus(HBV) load was determined by the serum level of HBV DNA.RESULTS:After adjusting for age,sex,and body mass index,we found that four linked single nucleotide polymorphisms(SNPs) of PNPLA3,including the rs738409 G allele(CHB + NAFLD group vs CHB group:odds ratio[OR]= 2.77,95%confidence interval[CI]:1.18-6.54;P = 0.02),rs3747206 T allele(CHB+ NAFLD group vs CHB group:OR = 2.77,95%CI:1.18-6.54;P = 0.02),rs4823173 A allele(CHB +NAFLD group vs CHB group:OR = 2.73,95%CI:1.16-6.44;P= 0.02),and rs2072906 G allele(CHB+ NAFLD group vs CHB group:OR = 3.05,95%CI:1.28-7.26;P = 0.01),conferred high risk to NAFLD in CHB patients.In patients with both CHB and NAFLD,these genotypes of PNPLA3 polymorphisms were associated with increased susceptibility to nonalcoholic steatohepatitis(NASH)(NAFLD activity score ≥3;P =0.01-0.03) and liver fibrosis(>1 Metavir grading;P =0.01-0.04).As compared to those with C/C and C/G at rs738409,C/C and C/T at rs3747206,G/G and G/A at rs4823173,and A/A and A/G at rs2072906,patients in the CHB + NAFLD group with G/G at rs738409,T/T at rs3747206,A/A at rs4823173,and G/G at rs2072906 showed significantly lower serum levels of HBV DNA(P< 0.01-0.05).CONCLUSION:Four linked SNPs of PNPLA3(rs738409,rs3747206,rs4823173,and rs2072906) are correlated with susceptibility to NAFLD,NASH,

关键词： Chronic hepatitis B Hepatitis B virus nonalcoholic fatty liver disease PNPLA3 Singlenucleotidepolymorphism

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Management of nonalcoholic fatty liver disease:An evidence-based clinical practice review

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World Journal of Gastroenterology 2014年第34期20卷 12182-12201页

作者： Juan P Arab Roberto Candia Rodrigo Zapata Cristián Mu?oz Juan P Arancibia Jaime Poniachik Alejandro Soza Francisco Fuster Javier Brahm Edgar Sanhueza Jorge Contreras M Carolina Cuellar Marco Arrese Arnoldo Riquelme Departamento de Gastroenterología Escuela de MedicinaPontificia Universidad Católica de Chile8330024 SantiagoChile Sección de Gastroenterología Hospital del SalvadorEscuela de MedicinaUniversidad de Chile7500922 SantiagoChile Unidad de Gastroenterología Universidad del DesarrolloClínica Alemana7650568 SantiagoChile Unidad de Trasplante Hepático Hospital del SalvadorUnidad de GastroenterologíaClínica Las Lilas7510524 SantiagoChile Sección de Gastroenterología Departamento de MedicinaHospital Clínico Universidad de ChileChile8380456 SantiagoChile Sección de Gastroenterología Hospital Clínico Universidad de ChileEscuela de MedicinaUniversidad de Chile8380456 SantiagoChile Digestive Diseases Unit Clínica Re?aca2540364 Vi?a del MarChile Gastroenterology Department Clínica Las Condes6770128 SantiagoChile Centre for Medical Education Escuela de MedicinaPontificia Universidad Católica de Chile8330024 SantiagoChile

AIM:To build a consensus among Chilean specialists on the appropriate management of patients with nonalcoholic fatty liver disease(NAFLD)in clinical practice.METHODS:NAFLD has now reached epidemic proportions worldwide.The optimal treatment for NAFLD has not been established due to a lack of evidence-based recommendations.An expert panel of members of the Chilean Gastroenterological Society and the Chilean Hepatology Association conducted a structured analysis of the current literature on NAFLD therapy.The quality of the evidence and the level of recommendations supporting each statement were assessed according to the recommendations of the United States Preventive Services Task Force.A modified three-round Delphi technique was used to reach a consensus among the experts.RESULTS:A group of thirteen experts was established.The survey included 17 open-ended questions that were distributed among the experts,who assessed the articles associated with each question.The levels of agreement achieved by the panel were 93.8%in the first round and 100%in the second and third rounds.The final recommendations support the indication of lifestyle changes,including diet and exercise,for all patients with NAFLD.Proven pharmacological therapies include only vitamin E and pioglitazone,which can be used in nondiabetic patients with biopsy-proven nonalcoholic steatohepatitis(the progressive form of NAFLD),although the long-term safety and efficacy of these therapies have not yet been established.CONCLUSION:Current NAFLD management is rapidly evolving,and new pathophysiology-based therapies are expected to be introduced in the near future.All NAFLD patients should be evaluated using a three-focused approach that considers the risks of liver disease,diabetes and cardiovascular events.

关键词： nonalcoholic fatty liver disease Steatosis Consens

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Mitochondrial DNA from hepatocytes as a ligand for TLR9: Drivers of nonalcoholic steatohepatitis?

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World Journal of Gastroenterology 2016年第31期22卷 6965-6971页

作者： Priya Handa Akhila Vemulakonda Kris V Kowdley Misael Uribe Nahum Méndez-Sánchez Senior Research Scientist Organ Care Research and Liver Care Network Swedish Medical Center Liver Research Unit Medica Sur Clinic and Foundation

nonalcoholic fatty liver disease(NAFLD) is the most common liver disease worldwide, affecting approximately one third of the Western world. It consists of a wide spectrum of liver disorders, ranging from fatty liver to nonalcoholic steatohepatitis(NASH), which consists of steatosis, ballooning injury and inflammation. Despite an alarming growth in the statistics surrounding NAFLD, there are as yet no effective therapies for its treatment. Innate immune signaling has been thought to play a significant role in initiating and augmenting hepatic inflammation, contributing to the transition from nonalcoholic fatty liver to NASH. An immune response is triggered by countless signals called damage-associated molecular patterns(DAMPs) elicited by lipid-laden and damaged hepatocytes, which are recognized by pattern recognition receptors(PRRs) on hepatic immune cells to initiate inflammatory signaling. In this editorial, in addition to summarizing innate immune signaling in NAFLD and discussing potential therapies that target innate immune pathways, we have described a recent study that demonstrated that mitochondrial DNA serves as a DAMP activating a hepatic PRR, TLR9, in mice and in the plasma of NASH patients. In addition to identifying a new ligand for TLR9 during NASH progression, the study shows that blocking TLR9 reverses NASH, paving the way for the development of future NASH therapy.

关键词： TLR9 Damage-associated molecular patterns Products of microbial metabolism produced by pathogens Mitochondrial DNA Hepatocytes nonalcoholic steatohepatitis nonalcoholic fatty liver disease Inflammation Innate immune signaling Kupffer cells Therapy

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miRNA-103: Molecular link between insulin resistance and nonalcoholic fatty liver disease

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World Journal of Gastroenterology 2015年第2期21卷 511-516页

作者： Qian Xu Ying Li Yong-Fang Shang Hui-Ling Wang Min-Xiu Yao Department of Endocrinology the Second Affiliated Hospital of Medical College of Qingdao University

AIM: To investigate the associations between miRNA-103(mi R-103) and insulin resistance and nonalcoholic fatty liver disease(NAFLD).METHODS: Serum samples were collected from 50 NAFLD patients who were overweight or obese(NAFLD group) and from 30 healthy subjects who served as controls(normal control group). Quantitative polymerasechain reaction was used to detect expression of mi R-103. Fasting plasma glucose, fasting insulin, and triglyceride(TG) levels were measured. Homeostasis model assessment was used to evaluate basal insulin resistance(HOMA-IR). Patient height and weight were measured to calculate body mass index(BMI).RESULTS: Compared with the normal control group, higher serum levels of mi R-103 were expressed in the NAFLD group(8.18 ± 0.73 vs 4.23 ± 0.81, P = 0.000). When P = 0.01(bilateral), mi R-103 was positively correlated with HOMA-IR(r = 0.881), TG(r = 0.774) and BMI(r = 0.878), respectively. mi R-103, TG and BMI were all independent factors for HOMAIR(β = 0.438/0.657/0.251, P = 0.000/0.007/0.001). mi R-103, TG, BMI and HOMA-IR were all risk factors for NAFLD(odds ratio = 2.411/16.196/1.574/19.11, P = 0.009/0.022/0.01/0.014).CONCLUSION: mi R-103 is involved in insulin resistance and NAFLD, and may be a molecular link between insulin resistance and NAFLD and a therapeutic target for these disorders.

关键词： mi RNA-103 Insulin resistance nonalcoholic fatty l

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Biomarkers in nonalcoholic fatty liver disease-the emperor has no clothes?

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World Journal of Gastroenterology 2015年第11期21卷 3223-3231页

作者： Madhusudana Girija Sanal Albert Einstein College of Medicine 1300 Morris Park AvenueBronxNY 10461United States

Fatty liver is present in over ten percentage of the world population and it is a growing public health problem.nonalcoholic fatty liver disease(NAFLD) is not a single disease,but encompasses a spectrum of diseases of different etiologies.It is difficult to find highly specific and sensitive diagnostic biomarkers when a disease is very complex.Therefore,we should aim to find relevant prognostic markers rather than accurate diagnostic markers which will help to minimize the frequency of liver biopsies to evaluate disease progression.There are several biomarker panels commercially available,however,there is no clear evidence that more sophisticated panels are better compared to simple criteria such as,presence of diabetes over five years,metabolic syndrome,obesity,obstructive sleep apnea,aspartate transaminase/alanine transaminase(ALT) ratio > 0.8 or ferritin levels > 1.5 times normal in patients with over six month history of raised ALT and/or ultrasonological evidence of fat in the liver.Currently the biomarker panels are not a replacement for a liver biopsy.However the need and benefit of liver biopsy in NAFLD is questionable because there is no convincing evidence that biopsy and detailed staging of NAFLD improves the management of NAFLD and benefits the patient.After all there is no evidence based treatment for NAFLD other than management of lifestyle and components of "metabolic syndrome".

关键词： nonalcoholic fatty liver disease Biomarkers Fibros

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Resveratrol and fenofibrate ameliorate fructose-induced nonalcoholic steatohepatitis by modulation of genes expression

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World Journal of Gastroenterology 2016年第10期22卷 2931-2948页

作者： Enas A Abd El-Haleim Ashraf K Bahgat Samira Saleh Department of Pharmacology and Toxicology Faculty of PharmacyCairo University Currently Faculty of Pharmaceutical Sciences and Pharmaceutical Industries Future University in Egypt

AIM: To evaluate the effect of resveratrol, alone and in combination with fenofibrate, on fructose-induced metabolic genes abnormalities in rats. METHODS: Giving a fructose-enriched diet(FED) to rats for 12 wk was used as a model for inducing hepatic dyslipidemia and insulin resistance. Adult male albino rats(150-200 g) were divided into a control group and a FED group which was subdivided into 4 groups, a control FED, fenofibrate(FENO)(100 mg/kg), resveratrol(RES)(70 mg/kg) and combined treatment( FENO + RES)( half the doses). A l l treatments were given orally from the 9th week till the end of experimental period. Body weight, oral glucose tolerance test(OGTT), liver index, glucose, insulin, insulin resistance(HOMA), serum and liver triglycerides(TGs), oxidative stress(liver MDA, GSH and SOD),serum AST, ALT, AST/ALT ratio and tumor necrosis factor-α(TNF-α) were measured. Additionally, hepatic gene expression of suppressor of cytokine signaling-3(SOCS-3), sterol regulatory element binding protein-1c(SREBP-1c), fatty acid synthase(FAS), malonyl Co A decarboxylase(MCD), transforming growth factor-β1(TGF-β1) and adipose tissue genes expression of leptin and adiponectin were investigated. Liver sections were taken for histopathological examination and steatosis area were determined.RESULTS: Rats fed FED showed damaged liver, impairment of glucose tolerance, insulin resistance, ox ida t ive s t re s s a nd dy s l ipide m ia. As fo r ge ne expression, there was a change in favor of dyslipidemia and nonalcoholic steatohepatitis(NASH) development. All treatment regimens showed some benefit in reversing the described deviations. Fructose caused deterioration in hepatic gene expression of SOCS-3, SREBP-1c, FAS, MDA and TGF-β1 and in adipose tissue gene expression of leptin and adiponectin. Fructose showed also an increase in body weight, insulin resistance(OGTT, HOMA), serum and liver TGs, hepatic MDA, serum AST, AST/ALT ratio and TNF-α compared to control. All treatments improved SOCS-3, FAS, MCD, TGF-β1 and leptin genes

关键词： Fructose nonalcoholic steatohepatitis Suppressor of cytokine signaling-3 Sterol regulatory element b

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