Impact of pre-operative weight loss on non-alcoholic fatty liver disease histopathology and insulin resistance in individuals undergoing bariatric surgery: a propensity matched cross-sectional comparison

ABSTRACT BACKGROUND: The effect of weight loss (WL) on histopathological aspects of non-alcoholic fatty liver disease (NAFLD) may provide further insights into the dynamics of hepatic recovery after WL. OBJECTIVE: To analyze the effects of pre-operative WL on insulin resistance- and NAFLD-related histology in individuals undergoing bariatric surgery (BS) with or without pre-operative WL. DESIGN AND SETTING: A matched cross-sectional study was conducted at a public university hospital and a private clinic in Campinas, Brazil. METHODS: An analytical, observational, cross-sectional study was conducted using prospectively collected databases of individuals who underwent BS and liver biopsy at either a public tertiary university hospital (with pre-operative WL) or a private clinic (without pre-operative WL). Random electronic matching by gender, age, and body mass index (BMI) was performed and two paired groups of 24 individuals each were selected. RESULTS: Of the 48 participants, 75% were female. The mean age was 37.4 ± 9.6. The mean BMI was 38.9 ± 2.6 kg/m2. Fibrosis was the most common histopathological abnormality (91.7%). Glucose was significantly lower in the WL group (92 ± 19.1 versus 111.8 ± 35.4 mg/dL; P = 0.02). Significantly lower frequencies of macrovesicular steatosis (58.3% versus 95.8%; P = 0.004), microvesicular steatosis (12.5% versus 87.5%; P < 0.001), and portal inflammation (50% versus 87.5%; P = 0.011) were observed in the WL group. CONCLUSION: Pre-operative WL was significantly associated with lower frequencies of macro- and mi- crovesicular steatosis, portal inflammation, and lower glycemia, indicating an association between the recent trajectory of body weight and histological aspects of NAFLD.

Macrophage heterogeneity role in NAFLD and NASH disease progression / 中华肝脏病杂志

Nonalcoholic fatty liver disease (NAFLD) is a type of metabolic stress liver injury that is closely associated with insulin resistance and genetic susceptibility. The continuum of liver injury in NAFLD can range from nonalcoholic fatty liver (NAFL) to nonalcoholic steatohepatitis (NASH) and even lead to cirrhosis and liver cancer. The pathogenesis of NAFLD is complicated. Pro-inflammatory cytokines, lipotoxicity, and gut bacterial metabolites play a key role in activating liver-resident macrophages (Kupffer cells, KCs) and recruiting circulating monocyte-derived macrophages (MoDMacs) to deposit fat in the liver. With the application of single-cell RNA-sequencing, significant heterogeneity in hepatic macrophages has been revealed, suggesting that KCs and MoDMacs located in the liver exert distinct functions in regulating liver inflammation and NASH progression. This study focuses on the role of macrophage heterogeneity in the development and occurrence of NAFLD and NASH, in view of the fact that innate immunity plays a key role in the development of NAFLD.

Effect of Xuanfuhua decoction on a mouse model of nonalcoholic steatohepatitis induced by high-fat, high-fructose, and high-cholesterol diet / 临床肝胆病杂志

Objective To investigate the intervention effect of Xuanfuhua decoction on mice with nonalcoholic steatohepatitis (NASH) induced by high-fat, high-fructose, and high-cholesterol diet. Methods A total of 32 male C57/BL6J mice were randomly divided into normal group, model group, Xuanfuhua decoction group, and obeticholic acid group, with 8 mice in each group. Since week 24 of modeling using high-fat, high-fructose, and high-cholesterol diet, each group was given the corresponding drug for intervention at a dose of 14.19 g/kg by gavage for the Xuanfuhua decoction group and 10 mg/kg by gavage for the obeticholic acid group and a volume of 20 mL/kg for gavage, once a day for 6 consecutive weeks. HE staining, oil red O staining, Sirius Red staining, and Masson staining were used to observe the pathological changes, lipid deposition, and collagen deposition of liver tissue; related kits were used to measure the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and glucose, as well as the content of TG and hydroxyproline (Hyp) in liver tissue; quantitative real-time PCR was used to measure the expression of genes associated with lipid metabolism, inflammation, and fibrosis in liver tissue; immunohistochemical staining was used to observe the positive expression of F4/80 and α-SMA in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the normal group, the model group had significant increases in body weight, liver wet weight, and serum levels of AST, ALT, TC, TG, LDL-C and glucose (all P < 0.01). HE staining showed hepatocyte steatosis, a large number of fat vacuoles, hepatocyte ballooning degeneration, and inflammatory cell infiltration in liver tissue of the mice in the model group, and the model group had a significant increase in NAFLD activity score (NAS) compared with the normal group ( P < 0.01). Oil red O staining showed the deposition of a large number of red lipid droplets with different sizes in hepatocytes of the mice in the model group, and compared with the normal group, the model group had significant increases in the area percentage of oil red O staining and the content of TG in the liver ( P < 0.01). Sirius Red staining and Masson staining showed significant collagen fiber hyperplasia in the perisinusoidal area, the central vein, and the portal area in the model group, and the model group had a significant increase in the content of Hyp in liver tissue compared with the normal group ( P < 0.05). Compared with the model group, the Xuanfuhua decoction group had significant reductions in the serum levels of AST, ALT, TC, TG, LDL-C, and glucose (all P < 0.05), significant improvements in hepatic steatosis, inflammatory infiltration, lipid droplet deposition, and collagen fiber hyperplasia, and significant reductions in NAS score, area percentage of oil red O staining, and content of TG and Hyp in the liver (all P < 0.05). Compared with the normal group, the model group had significant increases in the mRNA expression levels of lipid metabolism-related genes (SREBP-1c, FASN, SCD-1, PPAR-γ, and CD36), inflammation-related genes (F4/80, TNF-α, CCL2, and CD11b), and the fibrosis-related gene α-SMA (all P < 0.05), and immunohistochemical staining showed significant increases in the positive expression of F4/80 and α-SMA ( P < 0.01). Compared with the model group, the Xuanfuhua decoction group had significant reductions in the mRNA expression levels of SREBP-1c, FASN, SCD-1, PPAR-γ, CD36, F4/80, TNF-α, CCL2, CD11b, and α-SMA in liver tissue (all P < 0.05), and immunohistochemical staining showed significant reductions in the positive expression of F4/80 and α-SMA ( P < 0.01). Conclusion Xuanfuhua decoction has a good intervention effect on mice with NASH induced by high fat, high fructose, and high-cholesterol diet and can significantly inhibit hepatic lipid deposition, inflammatory response, and liver fibrosis.

Improvement effect and mechanism of different extracts from Tylophora yunnanensis on non-alcoholic steato-hepatitis / 中国药房

OBJECTIVE To investigate the improvement effect and mechanism of different extracts from Tylophora yunnanensis on non-alcoholic steatohepatitis （NASH）. METHODS Normal human liver LO2 cells were induced to steatosis by free fatty acid， then were divided into normal group， model group， silybin group （100 μmol/mL）， T. yunnanensis ethanol extracts （TYS） group （50 μg/mL）， T. yunnanensis ethyl acetate extracts （TYSA） group （50 μg/mL）， and T. yunnanensis n-butanol extracts （TYSB） group （50 μg/mL）. After 24 hours of drug intervention， the deposition of lipid droplets was observed in LO2 cells in each group. The contents of total cholesterol （TC）， triacylglycerol （TG）， malondialdehyde （MDA） and glutathione （GSH）， the activities of aspartate transaminase （AST）， alanine transaminase （ALT） and superoxide dismutase （SOD）， the mRNA expressions of Kelch-like ECH-associated protein 1（ Keap1）， nuclear factor E2-related factor 2（ Nrf2） and heme oxygenase 1（ HO- 1） were detected. NASH rat model was induced by a high-fat diet， and then divided into normal group， model group， silybin group （12.6 mg/kg）， TYS group （80 mg/kg）， TYSA group （80 mg/kg） and TYSB group （80 mg/kg）， with six rats in each group. The liver indexes of rats in each group were calculated after 6 weeks of drug intervention. The liver histopathological changes were observed， and the contents of TC， TG， HDL-C and LDL-C， AST and ALT activities in serum， the contents of MDA and GSH， SOD activities in liver tissue were detected. RESULTS Compared with model group， TYS， TYSA and TYSB could reduce lipid droplet deposition， intracellular TC， TG and MDA contents， AST and ALT activities， and increase SOD activity， GSH content， and Keap1， Nrf2， HO-1 mRNA expression levels in LO2 cells after steatosis to varying degrees， with some differences being statistically significant （P＜0.05）. They also significantly improved liver injury in NASH model rats， reduced their liver indexes， TC， TG， LDL-C and MDA contents， AST and ALT 1-042） activities， and increased HDL-C （except for TYS and TYSB）， GSH contents and SOD activity， with TYSA having the most significant effect （P＜0.05）. CONCLUSIONS TYS， TYSA and TYSB have a certain improvement effect on NASH， among which TYSA has the most obvious effect. Its mechanism of action may be related to upregulating the Keap1/Nrf2/HO-1 signaling pathway and inhibiting oxidative stress

Research progress of palmitoylation in non-alcoholic fatty liver disease and related liver diseases / 中国药科大学学报

@#Non-alcoholic fatty liver disease (NAFLD) has become a major public health hazard threatening human health worldwide.Yet, due to its complex pathogenesis, new drug development is difficult, with still insufficient clinical medication.Palmitoylation is a universal posttranslational modification of proteins catalyzed by palmitoyltransferase, affecting their stability, membrane localization and function.Recent studies have shown that palmitoylation is closely associated with NAFLD.This review summarizes the mechanisms of palmitoylation in NAFLD and analyzes the expression levels of the palmitoyltransferase family in liver tissues of NAFLD patients from GEO database, aiming to provide important clues to explore new mechanisms for NAFLD.

Risk stratifying non-alcoholic fatty liver disease patients for optimal care / 中华消化杂志

Non-alcoholic fatty liver disease (NAFLD) affects around 30% of the global adult population and is an important cause of cirrhosis and hepatocellular carcinoma. Compared with other chronic liver diseases, NAFLD is mostly seen by primary care physicians and non-hepatologists. Though the absolute number is huge, only a small fraction of patients will eventually develop liver-related complications. Therefore, it is important to use noninvasive tests wisely and develop a care model that involves not only hepatologists but also other colleagues seeing patients with NAFLD. With this background, the American Gastroenterological Association commissioned a multidisciplinary group to provide guidance on a clinical care pathway for identifying patients with advanced liver fibrosis due to NAFLD. The 4 key steps of this pathway include ① identifying patients at risk at primary care or non-hepatology settings, ② initial assessment with history taking and physical examination, ③ screening for advanced fibrosis using simple fibrosis score, and ④ specific fibrosis test such as vibration controlled transient elastography in patients with indeterminate fibrosis scores. This article discusses the rationale of the recommendations and highlights areas needing further data and refinement.

Predicting disruptions to drug pharmacokinetics and the risk of adverse drug reactions in non-alcoholic steatohepatitis patients

The liver plays a central role in the pharmacokinetics of drugs through drug metabolizing enzymes and transporters. Non-alcoholic steatohepatitis (NASH) causes disease-specific alterations to the absorption, distribution, metabolism, and excretion (ADME) processes, including a decrease in protein expression of basolateral uptake transporters, an increase in efflux transporters, and modifications to enzyme activity. This can result in increased drug exposure and adverse drug reactions (ADRs). Our goal was to predict drugs that pose increased risks for ADRs in NASH patients. Bibliographic research identified 71 drugs with reported ADRs in patients with liver disease, mainly non-alcoholic fatty liver disease (NAFLD), 54 of which are known substrates of transporters and/or metabolizing enzymes. Since NASH is the progressive form of NAFLD but is most frequently undiagnosed, we identified other drugs at risk based on NASH-specific alterations to ADME processes. Here, we present another list of 71 drugs at risk of pharmacokinetic disruption in NASH, based on their transport and/or metabolism processes. It encompasses drugs from various pharmacological classes for which ADRs may occur when used in NASH patients, especially when eliminated through multiple pathways altered by the disease. Therefore, these results may inform clinicians regarding the selection of drugs for use in NASH patients.

Fatores de risco para carcinoma hepatocelular em pacientes com doença gordurosa não alcóolica do fígado / Risk factors for hepatocellular carcinoma in patients with non-alcoholic fatty liver disease

ABSTRACT Non-alcoholic fatty liver disease is growing in worldwide prevalence and thus, is expected to have a higher number of NAFLD-related hepatocellular carcinoma (HCC) in the following years. This review describes the risk factors associated with HCC in NAFLD-patients. The presence of liver cirrhosis is the preponderant one. Male gender, PNPLA3 variants, diabetes, and obesity also appear to predispose to the development of HCC, even in non-cirrhotic subjects. Thus far, intensive lifestyle modifications, including glycemic control, and obesity treatment, are effective therapies for NAFLD/ non-alcoholic steatohepatitis and, therefore, probably, also for HCC. Some drugs that aimed at decreasing inflammatory activity and fibrosis, as well as obesity, were studied. Other data have suggested the possibility of HCC chemoprevention. So far, however, there is no definitive evidence for the routine utilization of these drugs. We hope, in the future, to be able to profile patients at higher risk of NAFLD-HCC and outline strategies for early diagnosis and prevention.

RESUMO A doença metabólica e doença hepática gordurosa metabólica estão aumentando a prevalência mundial e, portanto, espera-se um número maior de carcinoma hepatocelular (CHC) relacionado à doença hepática gordurosa não alcóolica (DHGNA) nos próximos anos. Esta revisão descreve os fatores de risco associados ao CHC em pacientes com DHGNA. A presença de cirrose hepática é a preponderante. Sexo masculino, variantes do gene PNPLA3, diabetes e obesidade também parecem predispor ao desenvolvimento de CHC, mesmo em indivíduos não cirróticos. Até agora, modificações significativas no estilo de vida, incluindo controle glicêmico e tratamento da obesidade, são terapias eficazes para DHGNA/ Esteatohepatite não-alcoolica e, portanto, provavelmente, também para CHC. Alguns medicamentos que propunham-se diminuir a atividade inflamatória e fibrose, bem como a obesidade, foram estudados. Outros dados sugeriram a possibilidade de quimioprevenção do CHC. Até o momento, no entanto, não há evidências definitivas para o uso rotineiro desses medicamentos. Esperamos, no futuro, poder traçar o perfil de pacientes com maior risco de DHGNA-CHC e traçar estratégias para diagnóstico precoce e prevenção.

IS HOMEOSTASIS MODEL ASSESSMENT FOR INSULIN RESISTANCE >2.5 A DISTINGUISHED CRITERIA FOR METABOLIC DYSFUNCTION-ASSOCIATED FATTY LIVER DISEASE IDENTIFICATION? / O modelo de avaliação da homeostase para resistência à insulina >2,5 é um critério distinto para a identificação da doença hepática gordurosa associada à disfunção metabólica?

ABSTRACT Background Insulin resistance (IR), assessed by different criteria, is an important factor in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). More recently with the characterization of this metabolic dysfunction-associated fatty liver disease (MAFLD), one of the proposed criteria for this diagnosis has been the determination of the homeostasis model assessment-insulin resistance (HOMA-IR). Objective: The purpose of this study was to evaluate the relationship of HOMA-IR>2.5 with clinical, metabolic, biochemical and histological data obtained in non-diabetic patients diagnosed with NAFLD by liver biopsy. Methods: Cross-sectional, retrospective study was carried out with data from 174 adult individuals of both genders with non-diabetics NAFLD, without obvious signs of portal hypertension. The body mass index (BMI) was classified according to the World Health Organization (1998), and the metabolic syndrome by the criteria of NCEP-ATP-III. Biochemical tests were evaluated using an automated method and insulinemia through immunofluorometric assay. Histological findings were classified according to Kleiner et al. (2005). Results: The mean age of the studied population was 53.6±11.2 years, with 60.3% being female. The average BMI was 30.3 kg/m2 and 75.9% of the patients had increased waist circumference. Among evaluated metabolic parameters, there was a higher prevalence of metabolic syndrome (MS) in patients with HOMA-IR>2.5, with no statistical difference in relation to BMI between studied groups. Values of liver enzymes and serum ferritin were significantly higher in patients with this marker of IR, who had a higher prevalence of non-alcoholic steatohepatitis (NASH) and advanced liver fibrosis. In the multivariate analysis, the clinical diagnosis of MS, hyperferritinemia and the presence of NASH in the liver biopsy were the factors independently associated with the presence of altered HOMA-IR. Conclusion: HOMA-IR values >2.5 identify patients with NAFLD with distinct clinical and metabolic characteristics and with a greater potential for disease progression, which validates this parameter in the identification of patients with MAFLD.

RESUMO Contexto A resistência à insulina (RI), avaliada por diferentes critérios, é um fator importante na patogênese da doença hepática gordurosa não alcoólica (DHGNA). Mas, recentemente, com a caracterização desta disfunção metabólica associada com a doença hepática gordurosa (DGH), um dos critérios propostos para este diagnóstico tem sido a determinação do modelo de avaliação da homeostase-resistência à insulina (HOMA-IR). Objetivo: O objetivo deste estudo foi avaliar a relação do HOMA-IR> 2,5 com dados clínicos, metabólicos, bioquímicos e histológicos obtidos em pacientes não diabéticos diagnosticados com DHGNA por biópsia hepática. Métodos Estudo transversal, retrospectivo, com dados de 174 indivíduos adultos de ambos os sexos com DHGNA não-diabética, sem sinais evidentes de hipertensão portal. O índice de massa corporal (IMC) foi classificado de acordo com a Organização Mundial da Saúde (1998) e a síndrome metabólica pelos critérios do NCEP-ATP-III. Os exames bioquímicos foram avaliados pelo método automatizado e a insulinemia por imunofluorometria. Os achados histológicos foram classificados de acordo com Kleiner et al. (2005). Resultados: A média de idade da população estudada foi de 53,6±11,2 anos, sendo 60,3% do sexo feminino. O IMC médio foi de 30,3 kg/m2 e 75,9% dos pacientes apresentaram circunferência da cintura aumentada. Entre os parâmetros metabólicos avaliados, houve maior prevalência de síndrome metabólica (SM) em pacientes com HOMA-IR >2,5, sem diferença estatística em relação ao IMC entre os grupos estudados. Os valores das enzimas hepáticas e da ferritina sérica foram significativamente maiores nos pacientes com este marcador de RI, que apresentaram maior prevalência de esteato-hepatite não alcoólica (EHNA) e fibrose hepática avançada. Na análise multivariada, o diagnóstico clínico de SM, hiperferritinemia e a presença de EHNA na biópsia hepática foram os fatores independentemente associados à presença de HOMA-IR alterado. Conclusão: Valores de HOMA-IR >2,5 identificam pacientes com DHGNA com características clínicas e metabólicas distintas e com maior potencial de progressão da doença, o que valida esse parâmetro na identificação de pacientes com DHG.

Liver cancer in Hidalgo State, Mexico: analysis of the status, risk factors and regional public health policy requirements: a cross-sectional correlational study

ABSTRACT BACKGROUND: In Latin America, liver cancer is one of the top causes of cancer mortality. It is the fifth most common cause of death among malignant tumors in Mexico and is the leading cause in Hidalgo State (43.8% of the population living in poverty). OBJECTIVE: To conduct a correlational analysis on the main risk factors for liver cancer in Hidalgo State, Mexico, including municipal disaggregation and comparison with the national level. DESIGN AND SETTING: Cross-sectional, correlational, descriptive and comparative epidemiological study using Mexican governmental databases covering 1990-2019. METHODS: A comprehensive review of the databases of the General Directorate of Health Information (DGIS) was performed to analyze official death figures, hospital discharges and national and municipal population projections, using specific search criteria defined in the Global Burden of Disease classification, based on the risk factors for liver cancer. RESULTS: Liver cancer rates showed an evident rise in Hidalgo (183%), moving from 21st place in Mexico in 1990 to 9th place in 2019. This increase was correlated with alcoholism. An increasing trend for liver cancer deaths, of 133.89%, is projected for 2030. Females and the population over 60 years of age are more affected. There are some critical regions with liver cancer death rates twice the national rate or more. CONCLUSION: Targeted effective public health strategies should be structured by identifying, characterizing and regionalizing critical marginalized municipalities that are vulnerable to alcoholism and other risk factors for liver cancer. This approach may be helpful for other states in Mexico or similar countries.

O risco de síndrome de apneia obstrutiva do sono em pacientes com doença hepática gordurosa não alcoólica está associado à obesidade e à presença de NASH / Obstructive sleep apnea syndrome risk in patients with non-alcoholic fatty liver disease is associated with obesity and presence of nash

ABSTRACT Background: Non-alcoholic fatty liver disease (NAFLD) is the most common form of liver disease and refers to a wide spectrum of histological abnormalities ranging from simple steatosis (HE) to non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis and hepatocellular carcinoma. Objective: To assess the risk of obstructive sleep apnea syndrome (OSAS) and relating it to demographic, biochemical and histological data in patients with non-alcoholic fatty liver disease. Methods: Cross-sectional cohort study in individuals with biopsy-proven NAFLD. Anthropometric and biochemical parameters, presence of metabolic syndrome and insulin resistance were evaluated. The Berlin Questionnaire (BQ) was applied to assess the risk of apnea and a food record was requested. Based on the BQ, participants were classified as high or low risk for OSAS. In the correlation of sleep apnea with the severity of NAFLD, presence of nonalcoholic steatohepatitis (NASH) and the degree of liver fibrosis were evaluated. Statistical analysis used the chi-square test, Student's t and bivariate logistic regression; values were expressed as mean ± standard deviation. This research project was approved by the Ethics Committee. Results: Regarding the parameters evaluated, significant differences were observed between the groups in terms of body mass index (BMI), waist and neck circumference. In the histological evaluation, patients classified as high risk were more likely to have fibrosis and NASH. In bivariate regression, the BMI, presence of fibrosis and steatohepatitis in the biopsy were independently associated with an elevated risk of the syndrome. Conclusion: A high prevalence of risk for OSAS was observed in the studied group, with a higher risk being independently associated with BMI and presence of steatohepatitis, suggesting that it is a factor associated with the severity of the disease.

RESUMO Contexto: A doença hepática gordurosa não alcoólica (DHGNA) é a forma mais comum de doença hepática e se refere a um amplo espectro de anormalidades histológicas que variam de esteatose simples a esteato-hepatite não alcoólica (EHNA), fibrose, cirrose e carcinoma hepatocelular. Objetivo: Avaliar o risco de síndrome da apneia obstrutiva do sono (SAOS) e relacioná-lo com dados demográficos, bioquímicos e histológicos em pacientes com doença hepática gordurosa não alcoólica. Métodos: Estudo de coorte transversal em indivíduos com DHGNA comprovada por biópsia. Foram avaliados parâmetros antropométricos e bioquímicos, presença de síndrome metabólica e resistência à insulina. O Questionário de Berlim (QB) foi aplicado para avaliar o risco de apneia e um registro alimentar foi solicitado. Com base no QB, os participantes foram classificados como de alto ou baixo risco para SAOS. Na correlação da apneia do sono com a gravidade da DHGNA, avaliou-se a presença de EHNA e o grau de fibrose hepática. Na análise estatística foram utilizados: o teste qui-quadrado, t de Student e regressão logística bivariada; os valores foram expressos como média ± desvio padrão. Este projeto de pesquisa foi aprovado pelo Comitê de Ética. Resultados: Em relação aos parâmetros avaliados, foram observadas diferenças significativas entre os grupos em relação ao índice de massa corporal (IMC), cintura e circunferência do pescoço. Na avaliação histológica, os pacientes classificados como de alto risco tiveram maior chance de apresentar fibrose e EHNA. Na regressão bivariada, o IMC, a presença de fibrose e esteato-hepatite na biópsia foram independentemente associados a um risco elevado da síndrome. Conclusão: Observou-se alta prevalência de risco para SAOS no grupo estudado, sendo o maior risco associado de forma independente ao IMC e à presença de esteato-hepatite, sugerindo que seja um fator associado à gravidade da doença.

A study of non-alcoholic fatty liver diseasein patients with type 2 diabetes mellitusand itscorrelation with cardiovascular risk

Background:NAFLD(non-alcoholic fatty liver disease) encompasses a variety of disorders of lipid metabolism ranging from NAFL, NASH to cirrhosis and rarely HCC. A great deal of evidence suggests that the metabolic syndrome predicts incident cardiovascular disease (CVD), so it is possible to hypothesize that NAFLD patients might portend a greater CVD risk and that NAFLD itself might confer a CVD risk above that associated with individual metabolic syndrome risk factors.Methods:55 T2DM patients were included in current study conducted fromAugust 2019 to September 2021 for assessment of NAFLD using USGabdomen, NAFLD fibrosis score and FIBROSCAN. ASCVD score was used for correlation between CVD risk and NAFLD.Results:Out of 55 patients 42 (76.4%) were having fatty liver based on USG abdomen while 13 (23.4%) patients were having no fatty liver. As far as steatosis is concerned mean CAP (dB/m) was 245±50.89 out of which 24 (43.6%) were having no or minimal steatosis (S0), while 31(56.4%) were having significant steatosis with 11 (20%), 9 (16.4%) and 11 (20%) having S1, S2 and S3 grade of steatosis respectively.15 patients out of 55 were of F0 grade while 19 (34.5%), 9 (16.4%), 7 (12.7%) and 5 (9.1%) were of grade F1, F2, F3 and F4 respectively. There was a positive statistically significant (p?0.001) association of ASCVD risk score with NAFLD fibrosis score and fibroscan: E (KPa). Conclusions:Our study found that the prevalence of NAFLD was quite higher in patients with T2DM based on both USG abdomen (76.4%) and transient elastography (steatosis-56.4% and fibrosis-65.5%) and a statistically significantassociation between fibrosis and ASCVD score with higher fibrosis associated with higher 10 years ASCVD score.

Effect of psoralen corylifolia on nonalcoholic steatohepatitis in young mice and its possible mechanism / 中国医师杂志

Objective:To study the protective effect and possible mechanism of psoralen corylifolia on non-alcoholic steatohepatitis (NASH) induced by high-fat diet in mice.Methods:The newly weaned female mice in the offspring of C57BL/6J mice fed with normal diet were selected as the control group (gavage of distilled water); the newly weaned female mice in the offspring of C57BL/6J mice fed with high-fat diet were randomly divided into model group (gavage distilled water), low-dose group[psoralen corylifolia 1.125 mg/(g·d)], high-dose group [psoralen corylifolia 2.25 mg/(g·d)] and vitamin E group [vitamin E 0.01 mg/(g·d)]. Six mice in each group were fed continuously for 8 weeks. Automatic biochemical analyzer was used to detect serum alanine aminotransferase (ALT), aspartate transaminase (AST), triglyceride (TG), total cholesterol (TC) level in mice; The liver tissue pathological changes were observed by hematoxylin-eosin (HE) and Sirius-red (SR) staining; The level of reactive oxygen species (ROS) in liver tissue was detected by dihydroethidium (DHE) fluorescence probe; the activity of NADPH oxidase was detected by kit; The protein expressions of nuclear factor-κB (NF-κB), phosphatidylinositol 3 kinase (PI3K p85), protein kinase B (Akt), P47 phox and protein kinase C-α (PKC-α) were detected by Western blot.Western blot. Results:The levels of serum ALT, AST, TG, TC and homeostasis model assessment of insulin resistance (HOMA-IR) index in the model group were higher than those in the control group (all P<0.01). After treatment, the levels of serum ALT, AST, TG, TC and HOMA-IR in low-dose group, high- dose group and vitamin E group were lower than those in model group (all P<0.05). HE and SR staining showed that hepatocytes in the model group were swollen, and there were lipid droplets of different sizes, vacuoles and obvious fibrosis. After treatment, hepatocyte steatosis and fibrosis decreased and the contents of ROS and NADPH oxidase in liver decreased(all P<0.05); Western blot showed that the p-p65/p65, p-Akt/Akt, p-PKC-α/PKC-α, PI3K, p85 and P47 phox protein expression in the model group were higher than those in the control group (all P<0.01). After treatment, the protein expression levels of p-p65/p65, p-Akt/Akt, p-PKC-α/PKC-α, PI3K, p85 and P47 phox decreased (all P<0.01). Among the above indexes, the protective effect of high-dose group on liver NASH was better than those of vitamin E group and low-dose group (all P<0.05). Conclusions:Psoralen corylifolia can improve the liver function of NASH model mice, which may be related to the inhibition of oxidative stress, inflammatory reaction and liver fibrosis, which provides a new idea for the prevention and treatment of children with NASH.

Effect of Quzhi Ruangan prescription on the farnesoid X receptor-fibroblast growth factor 19 pathway in rats with nonalcoholic steatohepatitis / 临床肝胆病杂志

Objective To investigate the effect of Quzhi Ruangan prescription on the farnesoid X receptor (FXR)-fibroblast growth factor 19 (FGF19) pathway in rats with nonalcoholic steatohepatitis (NASH). Methods Male Sprague-Dawley rats were randomly divided into normal group (Control group with 8 rats), model group (HFD group with 12 rats), simvastatin group with 8 rats, high-dose Quzhi Ruangan prescription group (QH group with 8 rats), and low-dose Quzhi Ruangan prescription group (QL group with 8 rats). The rats in the Control group were fed with a normal diet and those in the other groups were fed with a high-fat diet. Related samples were collected at the end of week 10 to observe liver pathological changes and measure the serum levels of liver function parameters, the level of FGF19 in the liver and the small intestine, and the level of bile acid (BA) in the liver. The expression levels of FXR in the small intestine and cholesterol 7α-hydroxylase (CYP7A1) in the liver were also measured. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t -test was used for further comparison bewteen two groups. Results Compared with the Control group, the HFD group showed the pathological manifestations of marked inflammatory lesions and steatosis. Compared with the HFD group, all administration groups had a significant increase in high-density lipoprotein cholesterol and significant reductions in alanine aminotransferase, aspartate aminotransferase, total cholesterol, triglyceride, and low-density lipoprotein cholesterol (all P < 0.05). Compared with the Control group, the HFD group had a significant reduction in FGF19 in the small intestine and a significant increase in BA in the liver (both P < 0.05). Compared with the HFD group, all administration groups had a significant increase in FGF19 in the small intestine and a significant reduction in BA in the liver (all P < 0.05). Compared with the Control group, the HFD group had a significant reduction in the mRNA expression of FXR in the small intestine and a significant increase in the mRNA expression of CYP7A1 in the liver (both P < 0.05). Compared with the HFD group, the QH group had a significant increase in the mRNA expression of FXR in the small intestine, while the QL group had a significant reduction (both P < 0.05), and the QH group had a significant reduction in the mRNA expression of CYP7A1 in the liver ( P < 0.05). Compared with the Control group, the HFD group had a significant reduction in the positive rate of FXR in the small intestine and a significant increase in the positive rate of CYP7A1 in the liver (both P < 0.05). Compared with the HFD group, the simvastatin group and the QH group had a significant increase in the positive rate of FXR in the small intestine (both P < 0.05), and the simvastatin group, the QH group, and the QL group had a significant reduction in the positive rate of CYP7A1 in the liver (all P < 0.05). Conclusion Quzhi Ruangan prescription can activate the FXR-FGF19 pathway in NASH rats and may exert a preventive and therapeutic effect on NASH through this pathway.

Application of farnesoid X receptor agonists in treatment of nonalcoholic steatohepatitis / 临床肝胆病杂志

Nonalcoholic steatohepatitis (NASH) is a liver disease with a relatively high prevalence worldwide and greatly threatens human health. In recent years, farnesoid X receptor-related drugs have played an important role in regulating the metabolism of bile acid, glucose, and lipids and inhibiting inflammation. This article summarizes the application of farnesoid X receptor agonists in the treatment of NASH, so as to provide a basis for the prevention and treatment of NASH.

Mechanism of Tibetan medicine Ershiwuwei Songshi Pills in regulating intestinal flora and improving non-alcoholic steatohepatitis / 中国中药杂志

This study aimed to investigate the effect of Tibetan medicine Ershiwuwei Songshi Pills(ESP) on the intestinal flora of non-alcoholic steatohepatitis(NASH) mice. Forty-eight male C57 BL/6 mice were randomly divided into the control group, model(methionine-choline-deficient, MCD) group, high-(0.8 g·kg~(-1)), medium-(0.4 g·kg~(-1)), and low-dose(0.2 g·kg~(-1)) ESP groups, and pioglitazone(PGZ, 10 mg·kg~(-1)) group, with eight mice in each group. Mice in the control group were fed with normal diet, while those in the remaining five groups with MCD diet for five weeks for inducing NASH. During modeling, they were gavaged with the corresponding drugs. The changes in body mass, daily water intake, and daily food intake were recorded. At the end of the experiment, the liver tissues were collected and stained with hematoxylin-eosin(HE) for observing the pathological changes, followed by oil red O staining for observing fat accumulation in the liver. The levels of serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) and triglyceride(TG) in liver tissue were measured. The changes in intestinal flora of mice were determined using 16 S rRNA high-throughput sequencing technology. The results showed that compared with the model group, the high-, medium-and low-dose ESP groups and the PGZ group exhibited significantly lowered AST and ALT in serum and TG in liver tissues and alleviated hepatocellular steatosis and fat accumulation in the liver. As demonstrated by 16 S rRNA sequencing, the abundance index and diversity of intestinal flora decreased in the model group, while those increased in the ESP groups. Besides, the Firmicutes to Bacteroidetes ratio decreased at the phylum level. In the alteration of the composition of intestinal flora, ESP reduced the abundance of Erysipelotrichia and Faecalibaculum but increased the abundance of Desulfovibrionaceae, Rikenellaceae, Lachnospiraceae, and Ruminococcaceae. This study has revealed that ESP has a protective effect against NASH induced by MCD diet, which may be related to its regulation of the changes in intestinal flora, alteration of the composition of intestinal flora, and inhibition of the intestinal dysbiosis.

Di'ao Xinxuekang alleviates non-alcoholic steatohepatitis in mice by up-regulating Nrf2/HO-1 signaling pathway / 中国中药杂志

The present study investigated the therapeutic effect and mechanism of Di'ao Xinxuekang(DXXK) on non-alcoholic steatohepatitis(NASH) in mice. Sixty-five C57 BL/6 J mice were randomly divided into a normal group and an experimental group for model induction with the high-fat diet for 16 weeks. Then the mice in the experimental group were randomly divided into a model group, an atorvastatin group(4 mg·kg~(-1)·d~(-1)), and high-(200 mg·kg~(-1)·d~(-1)), medium-(60 mg·kg~(-1)·d~(-1)), and low-dose(20 mg·kg~(-1)·d~(-1)) DXXK groups, with 10 mice in each group. Drugs were administered by gavage for eight weeks. Serum lipid, liver lipid, serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione reductase(GSH-Px) were determined. Interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay(ELISA). The liver index was calculated. The liver pathological change and lipid accumulation were observed by HE and oil red O staining. The liver ultrastructure was observed by the transmission electron microscope. The mRNA and protein expression of nuclear factor-erythroid 2 related factor 2(Nrf2) and heme oxygenase-1(HO-1) was detected by real-time fluorescence-based quantitative PCR and Western blot, respectively. The results showed that compared with the normal group, the model group displayed serum lipid and liver lipid metabolism disorders, elevated transaminase, lipid deposition, steatosis, and inflammation, suggesting that the NASH model in mice was properly induced. Compared with the model group, the DXXK groups showed decreased serum lipid, liver lipid, ALT, AST, MDA, IL-1β, and TNF-α, increased SOD and GSH-Px, alleviated hepatic steatosis, ballooning, and inflammation, and up-regulated Nrf2 and HO-1 gene and protein expression. In conclusion, DXXK can significantly alleviate NASH in mice, which is related to the inhibition of oxidative stress and inflammatory damage by up-regulating the Nrf2/HO-1 signaling pathway.

Association between serum sex hormone-binding globulin and non-alcoholic steatohepatitis / 中华内科杂志

Objective:To investigate the association between serum sex hormone-binding globulin (SHBG) and non-alcoholic steatohepatitis (NASH).Methods:In this cross-sectional study, a total of 371 middle-aged and young obese patients who were hospitalized and underwent liver puncture in Nanjing Drum Tower Hospital from January 2016 to April 2021 were included. The population was divided into control group ( n=43) and non-alcoholic fatty liver disease (NAFLD) group ( n=328) based on the non-alcoholic fatty liver disease activity score. Subjects in NAFLD group were further divided into non-alcoholic fatty liver (NAFL) ( n=60), uncertain-NASH ( n=172), and NASH ( n=96). Serum SHBG was tested in patients with NAFLD who were divided into three subgroups according to tertiles. The liver pathological characteristics in different SHBG level subgroups were compared. The risk factors of NASH were analyzed by logistic regression. The prediction model of NASH noninvasive diagnosis was established by forward stepwise regression, and the diagnostic value of non-invasive model for NASH was evaluated by receiver operating characteristic (ROC) curve. Results:The median age in patients were (32±10) years old with a body mass index of (39.16±6.58) kg/m2, including 236 females (63.6%). Serum SHBG level [ M ( Q1, Q3)] in NAFLD group was significantly lower than that in control group [16.90 (11.43, 23.00) vs. (23.45 (15.40, 31.22) mmol/L, P<0.05], and progressively diminished in NAFL, uncertain-NASH and NASH subgroups [(22.24±10.47), (20.57±19.58), (15.80±8.74) mmol; P for trend<0.05]. Compared with the high-leveled SHBG subgroup, the steatosis score (2.09±0.80 vs. 1.51±0.72, P<0.01) and lobular inflammation score (1.10±0.68 vs. 0.85±0.68, P<0.05) were significantly higher in the low-leveled SHBG group. Multivariate logistic regression analysis indicated that lower serum SHBG level was an independent risk factor for NASH ( OR=2.527, 95% CI: 1.296 to 4.928, P<0.05). The area under ROC curve of SHBG combined with aspartate aminotransferase in predicting NASH in NAFLD patients was 0.752 (95% CI: 0.696 to 0.809). Conclusion:Low serum SHBG level is associated with NASH.

Clinical-biochemical and Fibroelastometric Results of the Mare's Milk Application in Non-Alcoholic Steathepatitis / Resultados clínico-bioquímicos y fibroelastométricos de la aplicación de la leche de yegua en la esteatohepatitis no alcohólica

Introduction: Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the world, representing the hepatic component of metabolic syndrome, and currently affects an average of 20-33 percent of the adult population. Non-alcoholic steatohepatitis (NASH) is a severe form of NAFLD and is a consequence of overweight, obesity, and metabolic syndrome. Objective: The study aimed to evaluate the effectiveness of the use of freeze-dried mare's milk in NASH. Methods: Clinical-biochemical, ultrasound, and fibroelastometric research methods were used in NASH patients receiving mare's milk in comparison with the control groups. Results: The result of the study demonstrates noticeable gain not only in the clinical symptoms of the disease but also in the laboratory and instrumental indicators, as well as health improvement and a decrease in symptoms of concomitant pathology. Taking mare's milk not only normalized liver biochemical parameters but also decreased cholesterol metabolism (total cholesterol, LDL, TG), the degree of liver steatosis, and existing hepatomegaly declined too. This pleiotropic effect of mare's milk points to the pathogenetic feasibility for the use of Saumal in NAFLD, including NASH. Conclusions: Addressing the problems of healthy nutrition in NASH, mare's milk can be considered as a pathogenetically justified, highly effective, and affordable natural therapeutic and prophylactic agent. The value of the product is determined by its multicomponent balanced qualitative composition. Hepatoprotective, hypocholesterolemic, and lipotropic properties of mare's milk in this pathology are marked(AU)

Introducción: La enfermedad del hígado graso no alcohólico es la enfermedad hepática más común en el mundo, representa el componente hepático del síndrome metabólico y actualmente afecta a un promedio de 20 a 33 por ciento de la población adulta. La esteatohepatitis no alcohólica es una forma grave de hígado graso no alcohólico y es una consecuencia del sobrepeso, la obesidad y el síndrome metabólico. Objetivo: Evaluar la efectividad del uso de leche de yegua liofilizada en hígado graso no alcohólico. Métodos: Se utilizaron métodos de investigación clínico-bioquímicos, ecográficos y fibroelastométricos en pacientes con hígado graso no alcohólico que recibieron leche de yegua en comparación con los grupos de control. Resultados: El resultado del estudio demuestra una ganancia notable no solo en los síntomas clínicos de la enfermedad, sino también en los indicadores de laboratorio e instrumentales, así como una mejora de la salud y una disminución de los síntomas de la enfermedad concomitante. La ingesta de leche de yegua no solo normalizó los parámetros bioquímicos del hígado, sino que también disminuyó el metabolismo del colesterol (colesterol total, LDL, TG), el grado de esteatosis hepática y la hepatomegalia existente también disminuyeron. Este efecto pleiotrópico de la leche de yegua apunta a la viabilidad patogénica del uso de Saumal en hígado graso no alcohólico, incluida la esteatohepatitis no alcohólica. Conclusión: Al abordar los problemas de una nutrición saludable en la enfermedad hígado graso no alcohólico, la leche de yegua se puede considerar como un agente terapéutico y profiláctico natural patogénicamente justificado, altamente efectivo y asequible. El valor del producto está determinado por su composición cualitativa equilibrada multicomponente. Las propiedades hepatoprotectoras, hipocolesterolémicas y lipotrópicas de la leche de yegua en esta enfermedad son marcadas(AU)

Vitamina B12 e homocisteína em pacientes com DHGNA: revisão sistemática e metanálise / Vitamin b12 and homocysteine levels in patients with nafld: a systematic review and metanalysis

ABSTRACT BACKGROUND: The vitamin B12 absorption can be affected in patients with nonalcoholic fatty liver disease (NAFLD), and low serum vitamin B12 levels has been related to the high homocysteine (HCY) levels and to the degree of NAFLD. OBJECTIVE: To carry out a systematic review and metanalysis of serum vitamin B12 and HCY levels in patients with NAFLD. METHODS: Original studies including serum vitamin B12 and HCY levels in humans with NAFLD were included. The searches were performed in four databases. RESULTS: 159 studies were identified, and after excluding the duplicates and non-eligible titles, eight original articles were included. Six out of eight showed higher B12 levels in NAFLD patients (404.9±136.2 pg/mL in relation to controls 353.91±117.3 pg/mL). Seven of the eight studies also showed higher HCY levels in NAFLD patients (14.2±3.44 umol/L in relation to controls 11.05±3.6 umol/L). The results for serum vitamin B12 and HCY levels were submitted to metanalysis, showing no difference in the vitamin B12 levels between patients with NAFLD and controls. However, the levels of Hcy were higher in NAFLD patients than in controls. CONCLUSION: There was no relashionship between the vitamin B12 levels and NAFLD. The levels of HCY were significantly higher in patients with NAFLD, suggesting this could be a potential marker for liver damage.

RESUMO CONTEXTO: A absorção de vitamina B12 pode ser afetada em pacientes com doença hepática gordurosa não alcoólica (DHGNA), e baixos níveis séricos de vitamina B12 têm sido relacionados a níveis elevados de homocisteína (HCI) ao grau de DHGNA. OBJETIVO: Realizar revisão sistemática e metanálise dos níveis séricos de vitamina B12 e de HCI em pacientes com DHGNA. MÉTODOS: Estudos originais que incluíssem avaliação dos níveis séricos de vitamina B12 e de HCI em humanos com DHGNA foram incluídos. As buscas foram realizadas em quatro bases de dados. RESULTADOS: Foram identificados 159 estudos e, após exclusão das duplicatas e dos não elegíveis, oito artigos originais foram incluídos. Seis dos oito artigos apresentaram níveis mais elevados de vitamina B12 nos pacientes com DHGNA (404,9±136,2 pg/mL) em relação aos controles (353,91±117,3 pg/mL). Sete dos oito estudos determinaram os níveis de HCI, estando aumentados em pacientes com DHGNA (14,2±3,44 umol/L) em relação aos controles (11,05±3,6 umol/L). Os resultados dos níveis séricos de vitamina B12 e HCI foram submetidos à metanálise, mostrando que não há diferença nos níveis de vitamina B12 entre os pacientes com DHGNA e os controles. No entanto, os níveis de HCI foram maiores nos pacientes com DHGNA do que nos controles. CONCLUSÃO: Não houve relação entre DHGNA e nível sérico de vitamina B12. Os níveis de HCI foram significativamente maiores em pacientes com DHGNA, sugerindo que esse poderia ser um potencial marcador de lesão hepática.