Hepatic steatosis is associated with dysregulated cholesterol metabolism and altered protein acetylation dynamics in chickens

Background Hepatic steatosis is a prevalent manifestation of fatty liver, that has detrimental effect on the health and productivity of laying hens, resulting in economic losses to the poultry industry. Here, we aimed to systematically investigate the genetic regulatory mechanisms of hepatic steatosis in laying hens.Methods Ninety individuals with the most prominent characteristics were selected from 686 laying hens according to the accumulation of lipid droplets in the liver, and were graded into three groups, including the control, mild hepatic steatosis and severe hepatic steatosis groups. A combination of transcriptome, proteome, acetylome and lipidome analyses, along with bioinformatics analysis were used to screen the key biological processes, modifications and lipids associated with hepatic steatosis.Results The rationality of the hepatic steatosis grouping was verified through liver biochemical assays and RNA-seq. Hepatic steatosis was characterized by increased lipid deposition and multiple metabolic abnormalities. Integration of proteome and acetylome revealed that differentially expressed proteins(DEPs) interacted with differentially acetylated proteins(DAPs) and were involved in maintaining the metabolic balance in the liver. Acetylation alterations mainly occurred in the progression from mild to severe hepatic steatosis, i.e., the enzymes in the fatty acid oxidation and bile acid synthesis pathways were significantly less acetylated in severe hepatic steatosis group than that in mild group(P < 0.05). Lipidomics detected a variety of sphingolipids(SPs) and glycerophospholipids(GPs) were negatively correlated with hepatic steatosis(r ≤-0.5, P < 0.05). Furthermore, the severity of hepatic steatosis was associated with a decrease in cholesterol and bile acid synthesis and an increase in exogenous cholesterol transport.Conclusions In addition to acquiring a global and thorough picture of hepatic steatosis in laying hens, we were able to reveal the role of acetylation in hepatic steatosis and depict the changes in hepatic cholesterol metabolism. The findings provides a wealth of information to facilitate a deeper understanding of the pathophysiology of fatty liver and contributes to the development of therapeutic strategies.

Evaluation of Hepatic Fibrosis and Hepatic Steatosis by Pulse Elastography (FIBROSCAN/CAP) in Asymptomatic Patients about 170 Cases at the Donka CHU National Hospital in Conakry

Introduction: Fibroscan is a recent, non-invasive and non-irradiating diagnostic method. It is based on the principle of ultrasound, which enables liver tissue elasticity to be quantified using a probe, and fibrosis to be assessed. Fibroscan measures both elasticity correlated with hepatic fibrosis and CAP correlated with steatosis. The aim of this study was to evaluate hepatic fibrosis and steatosis using pulse elastometry (Fibroscan/CAP). Methods: This was a descriptive and analytical cross-sectional study in which 170 patients were included. It was conducted from October 1 2021 to December 31 2023, i.e. 27 months, in an outpatient clinic in the hepato-gastroenterology department of the Donka national hospital of the CHU Conakry. Results: Of the 170 patients identified, 87 were male (51%) and 83 female (49%), giving a M/F sex ratio of 1.04. The average age of our patients was 40. The 30 - 50 age group was the most affected, with a frequency of 58.23% (n = 99), followed by the 50 age group with a frequency of 29.41% (n = 50). Hepatomegaly, steatotic liver on ultrasonography, transaminase elevation and obesity were the main indications, respectively: (21.76%), (17.65%), (14.71%), and (13.53%). The examinations were requested by hepatogastroenterologists (47.06%), diabetologists (35.88%) and general practitioners (29%). Of the 170 patients, 100 patients (58.82%) had no significant fibrosis F0F1, 39 (22.94%) had moderate fibrosis F2, 20 patients (11.76%) had severe fibrosis F3 and 11 patients (6.47%) had fibrosis F4. Hepatic steatosis: 62 patients (36.47%) had no S0 steatosis;29.41% had S1 steatosis, 20% had S2 steatosis and 24 patients (14.11%) had S3 steatosis. Abdominal ultrasound revealed a normal liver in 67.05% of patients, hepatic steatosis in 29.41% and non-decompensated cirrhosis in 6 cases. Thus, 108 patients had the parameters required to calculate the Fatty Liver Index (FLI), steatosis was present in 20% of our patients, while 29.41% had an undetermined status and 24 14.11% had a normal FLI. Conclusion: Identifying subjects at risk of metabolic steatopathy, diagnosing and managing these patients is a public health issue and one of the future challenges of hepato-gastroenterology. Fibroscan is an increasingly popular screening tool for hepatic fibrosis and steatosis. The fight against obesity must be a priority.

Theasinensin A attenuated diabetic development by restoring glucose homeostasis, improving hepatic steatosis and modulating gut microbiota in high-fat-diet/streptozotocin-induced diabetic mice

Theasinensin A(TSA),a dimer of epigallocatechin gallate,has been preliminarily demonstrated to have hypoglycemia and anti-inflammatory effects.However,little information is available on its potential mechanisms of anti-diabetes.Therefore,the present study aimed to investigate the influence of TSA on glucose and lipid metabolism and gut microbiota in high-fat-diet/streptozotocin-induced diabetic mice.As result,TSA improved polydipsia,polyphagia and impaired glucose tolerance of diabetic mice,declined the fasting blood glucose and hepatic triglyceride level,and enhanced the expression at mRNA level of insulin receptor substrate,phosphoinositide 3-kinase,protein kinase B and glucagon-like peptide 1 receptor(GLP-1R)in the diabetic liver.Moreover,TSA could restore the disorder of gut microbiota of diabetic mice.High-dose(100 mg/kg)TSA showed better benefi cial effects from the blood biochemical parameters,hepatic function and gut microbiota.In general,high-dose TSA significantly modulated gut microbiota by increasing the relative abundance of Akkermansia and decreasing the relative abundances of Acetatifactor,Anaerotruncus,Pseudofl avonifactor,Oscillibacter and Clostridium clusters.The results indicated that TSA could exert an anti-diabetes effect in diabetic mice through restoring glucose homeostasis,declining hepatic steatosis,activating insulin and GLP-1 signaling pathways,and ameliorating gut microbiota dysbiosis.

Two-point Dixon and six-point Dixon magnetic resonance techniques in the detection,quantification and grading of hepatic steatosis

BACKGROUND Hepatic steatosis is a very common problem worldwide.AIM To assess the performance of two-and six-point Dixon magnetic resonance(MR)techniques in the detection,quantification and grading of hepatic steatosis.METHODS A single-center retrospective study was performed in 62 patients with suspected parenchymal liver disease.MR sequences included two-point Dixon,six-point Dixon,MR spectroscopy(MRS)and MR elastography.Fat fraction(FF)estimates on the Dixon techniques were compared to the MRS-proton density FF(PDFF).Statistical tests used included Pearson’s correlation and receiver operating characteristic.RESULTS FF estimates on the Dixon techniques showed excellent correlation(≥0.95)with MRS-PDFF,and excellent accuracy[area under the receiver operating characteristic(AUROC)≥0.95]in:(1)Detecting steatosis;and(2)Grading severe steatosis,(P<0.001).In iron overload,two-point Dixon was not evaluable due to confounding T2*effects.FF estimates on six-point Dixon vs MRS-PDFF showed a moderate correlation(0.82)in iron overload vs an excellent correlation(0.97)without iron overload,(P<0.03).The accuracy of six-point Dixon in grading mild steatosis improved(AUROC:0.59 to 0.99)when iron overload cases were excluded.The excellent correlation(>0.9)between the Dixon techniques vs MRSPDFF did not change in the presence of liver fibrosis(P<0.01).CONCLUSION Dixon techniques performed satisfactorily for the evaluation of hepatic steatosis but with exceptions.

Prevalence and Factors Associated with Hepatic Steatosis in Patients with Metabolic Syndrome in Cameroon: Cases of 4 Reference Hospitals

Introduction: Nonalcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease worldwide and its prevalence increases with that of metabolic syndrome and its components. NAFLD is associated with complications such as cirrhosis and hepatocellular carcinoma. Diagnosis is mainly based on liver biopsy, but there are validated non-invasive methods. The purpose of the study was to assess the impact of metabolic steatopathy in patients with metabolic syndrome in Cameroon. Methods: This was a cross-sectional and analytical study conducted over a 6-month period from January 1st, 2019, to August 31st, 2022. Included were patients with metabolic syndrome who had consulted in endocrinology or gastroenterology at Yaoundé Central Hospital, Douala General Hospital and Douala Gyneco-obstetric and Pediatric Hospital. The diagnosis of NAFLD was made on abdominal ultrasound in front of a homogeneous or heterogeneous hyperechogenic aspect of the hepatic parenchyma compared to that of the right renal cortex called “brilliant liver” and fibrosis evaluated through non-invasive scores (Fib4 and NALFD Fibrosis score). Logistic regression by a uni- and multivariate analysis made it possible to search for the associated factors. Results. We included 133 patients. The female sex represented 64.7%. The mean age was 55 ± 9 years. The prevalence of NAFLD was 48.9%. At the evaluation of fibrosis was significant according to FIB-4 and NAFLD fibrosis score respectively in 6.2% and 4.6% of cases. The independently associated factors were Triglyceridemia ≤ 1.5 g/l (OR = 0.33;95% CI [0.11 - 0.95];p = 0.04) and LDL hypercholesterolemia (OR = 2.94;95% CI [1.07 - 8.11];p = 0.036). Conclusion: NAFLD was present in almost half of patients with metabolic syndrome. We had very few patients with significant fibrosis, but it needs to be further evaluated. The associated factors are hypertriglyceridemia and LDL hypercholesterolemia.

Hepatic steatosis,low-grade chronic inflammation and hormone/growth factor/adipokine imbalance

Non-alcoholic fatty liver disease (NAFLD), a further expression of metabolic syndrome, strictly linked to obesity and diabetes mellitus, is characterized by insulin resistance (IR), elevated serum levels of free fatty acids and fatty infi ltration of the liver, which is known as hepatic steatosis. Hepatocyte apoptosis is a key feature of this disease and correlates with its severity. Free-fatty-acidinduced toxicity represents one of mechanisms for the pathogenesis of NAFLD and hormones, growth factors and adipokines influence also play a key role. This review highlights the various pathways that contribute to the development of hepatic steatosis. Circulating concentrations of inflammatory cytokines are reckoned to be the most important factor in causing and maintaining IR. Low-grade chronic inflammation is fundamental in the progression of NAFLD toward higher risk cirrhotic states.

Evaluation of controlled attenuation parameter in assessing hepatic steatosis in patients with autoimmune liver diseases

BACKGROUND Hepatic steatosis commonly occurs in some chronic liver diseases and may affect disease progression.AIM To investigate the performance of controlled attenuation parameter(CAP)for the diagnosis of hepatic steatosis in patients with autoimmune liver diseases(AILDs).METHODS Patients who were suspected of having AILDs and underwent liver biopsy were consistently enrolled.Liver stiffness measurement(LSM)and CAP were performed by transient elastography.The area under the receiver operating characteristic(AUROC)curve was used to evaluate the performance of CAP for diagnosing hepatic steatosis compared with biopsy.RESULTS Among 190 patients with biopsy-proven hepatic steatosis,69 were diagnosed with autoimmune hepatitis(AIH),18 with primary biliary cholangitis(PBC),and 27 with AIH-PBC overlap syndrome.The AUROCs of CAP for the diagnosis of steatosis in AILDS were 0.878(0.791-0.965)for S1,0.764(0.676-0.853)for S2,and 0.821(0.716-0.926)for S3.The CAP value was significantly related to hepatic steatosis grade(P<0.001).Among 69 patients with AIH,the median CAP score was 205.63±47.36 dB/m for S0,258.41±42.83 dB/m for S1,293.00±37.18 dB/m for S2,and 313.60±27.89 dB/m for S3.Compared with patients with nonalcoholic fatty liver disease(NAFLD)presenting with autoimmune markers,patients with AIH concomitant with NAFLD were much older and had higher serum IgG levels and LSM values.CONCLUSION CAP can be used as a noninvasive diagnostic method to evaluate hepatic steatosis in patients with AILDs.Determination of LSM combined with CAP may help to identify patients with AIH concomitant with NAFLD from those with NAFLD with autoimmune phenomena.

Novel insights into the effect of Xiaoyao san on corticosterone-induced hepatic steatosis:inhibition of glucocorticoid receptor/perilipin-2 signaling pathway

Objective:Xiaoyao san(XYS)is a classic traditional Chinese medicinal formula.It has been clinically administered to regulate liver function.However,its mechanisms in glucocorticoid-induced hepatic steatosis are unknown.This study aimed to investigate whether XYS protects against corticosterone(CORT)-induced hepatic steatosis,and to explore its mechanism.Methods:High-fat diet mice induced with hepatic steatosis by 2mg/kg CORT were administered 2.56 g/kg or 5.12 g/kg XYS daily for 7 weeks.The effects of XYS on hepatic steatosis in mice were evaluated by H&E and Oil Red O staining and by measuring their plasma lipids(triglyceride,total cholesterol,and free fatty acids).The mechanism of XYS against hepatic steatosis was investigated by network pharmacology,immunohistochemistry,western blotting,and gain-of-function/loss-offunction experiments.Results:XYS alleviated CORT-induced steatosis,decreased plasma lipids,and inhibited glucocorticoid receptor(GR)activation in the liver.Network pharmacology data indicated that XYS may have mitigated hepatic steatosis via GR which mediated adipose differentiation-related protein(ADFP).Gain-of-function/loss-of-function experiments in vitro confirmed that GR positively regulated ADFP expression.Conclusions:XYS ameliorated CORT-induced hepatic steatosis by downregulating the GR/ADFP axis and inhibiting lipid metabolism.Our studies implicate that XYS is promising as a therapy for CORT-induced hepatic steatosis,and lay the foundation for designing novel prophylactic and therapeutic strategies on CORT-induced hepatic steatosis.

Is hepatic steatosis associated with left ventricular mass index increase in the general population?

AIM To investigate the association between hepatic steatosis and change in left ventricular mass index(LVMI) over five years, and examine whether systolic and diastolic blood pressures are mediators of the association between hepatic steatosis and LVMI using a general population sample.METHODS We analyzed data from the Study of Health in Pomerania. The study population comprised 1298individuals aged 45 to 81 years. Hepatic steatosis was defined as the presence of a hyperechogenic pattern of the liver together with elevated serum alanine transferase levels. Left ventricular mass was determined echocardiographically and indexed to height2.7. Path analyses were conducted to differentiate direct and indirect paths from hepatic steatosis to LVMI encompassing systolic and diastolic blood pressure as potential mediating variables.RESULTS Hepatic steatosis was a significant predictor for all measured echocardiographic characteristics at baseline. Path analyses revealed that the association of hepatic steatosis with LVMI change after five years was negligibly small(β =-0.12, s.e. = 0.21, P = 0.55). Systolic blood pressure at baseline was inversely associated with LVMI change(β =-0.09, s.e. = 0.03, P < 0.01), while no association between diastolic blood pressure at baseline and LVMI change was evident(β = 0.03, s.e. = 0.05, P = 0.56). The effect of the indirect path from hepatic steatosis to LVMI via systolic baseline blood pressure was small(β =-0.20, s.e. = 0.10, P = 0.07). No indirect effect was observed for the path via diastolic baseline blood pressure(β = 0.03, s.e. = 0.06, P = 0.60). Similar associations were observed in the subgroup of individuals not receiving beta-blockers, calcium channel blockers, or drugs acting on the reninangiotensin system.CONCLUSION Baseline associations between hepatic steatosis and LVMI do not extend to associations with LVMI change after five years. More studies are needed to study the longitudinal effects of hepatic steatosis on LVMI.

Hepatic steatosis with mass effect:A case report

BACKGROUND Hepatic steatosis is a common radiologic finding.Some imaging inklings are the absence of a mass effect,and there is currently no report of hepatic steatosis with mass effect.CASE SUMMARY A 23-year-old female was admitted due to a liver mass for half a month.No obvious abnormalities were found in physical and laboratory examinations.Ultrasound,computed tomography,and magnetic resonance imaging showed a huge mass between the liver and stomach with a significant mass effect,and the caudate lobe and left lobe of the liver were involved.The signal on T2-and T1-weighted fat-saturated images of the mass was significantly reduced,and the enhanced scan showed inhomogeneous enhancement.Surgical and pathological findings indicated the diagnosis of hepatic steatosis.The operation and re-review of the patient’s images showed that the lesion was supplied by the branch of the hepatic artery.The signal on T1-weighted out-of-phase images of the lesion was lower than on in-phase images,and there was no black rim cancellation artifact around the hepatic steatosis area on T1-weighted out-of-phase images.The dynamic enhancement pattern of the lesion was similar to that of the adjacent normal liver parenchyma.The above characteristics suggested that the lesion was hepatic steatosis.However,in this case,the lesion showed exogenous growth and was mass-like,with an obvious mass effect,which has not been reported previously.CONCLUSION Hepatic steatosis could grow exogenously and has an obvious mass effect.It needs to be distinguished from fat-rich tumors.The T1-weighted in-and out-of-phase images and dynamic enhanced scanning are valuable for differential diagnosis of this lesion.

In vivo percutaneous reflectance spectroscopy of fatty liver development in rats suggests that the elevation of the scattering power is an early indicator of hepatic steatosis

This study assessed whether there was a scat tering spectral mar ker quantifiable by reflectance measurements that could indicate early development of hepatic steatosis in rats for potential applications to pre procurement organ evaluation.Sixteen rats were fed a methionine choline-deficient(MCD)diet and eight rats were fed a normal diet.Direct assessment of the liver parenchyma of rats in vivo was performed by percut aneous reflect ance spectroscopy using a single fiber probe at the beginning of diet-intake and arbitrary post-diet-intake times up to 11 weeks to render longitudinal comparison.Histological sampling of the liver over the duration of diet adm inistration was performed on two MCD diet treated rats and one control rat eutha-nized after reflectance spectroscopy measurement.The images of hematoxylin/eosin-stained liver specimens were analyzed morphometrically to evahuate the lipid size changes associated with the level of steatosis.The MCD-diet-treated group(n=16)had mild steatosis in seven rats,moderate in three rats,severe in six rats,and no other significant pathology.No control rats(n=8)developed hepatic steatosis.Among the parameters retrieved from per-SfS,only the scat tering power(can be either positive or negative)appeared to be statist ically diferent between MCD-treated and control livers.The scattering power for the 16 MCD-diet-treated livers at the time of euthanasia and presenting various levels of steatosis was 033±0.21,in comparison to 0.036±0.25 of the eight control livers(p=0.0189).When evaluated at days 12 and 13 combined,the scattering power of the 16 MCD-diet-treated livers was 032±0.17,in comparison to 0.10±0.11 of the eight control livers(p=0.0017).All of four MCD-treated livers harvested at days 12 and 13 presented mild steatosis with sub-micron size lipid droplets,even though none of the MCD-treated livers were sonogr aphically remarkable for fatty changes.The elevation of the scattering power may be a valuable marker indicating early hepatic steatosis before the steatosis is sonographically detectable.

Testosterone therapy reduces hepatic steatosis in men with type 2 diabetes and low serum testosterone concentrations

BACKGROUND Non-alcoholic fatty liver disease(NAFLD)is highly prevalent in people with diabetes with no available treatment.AIM To explore the effect of testosterone treatment on liver.Testosterone therapy improves insulin resistance and reduces total body fat,but its impact on the liver remains poorly studied.METHODS This secondary analysis of a 40 wk,randomised,double-blinded,placebocontrolled trial of intramuscular testosterone undecanoate in men with type 2 diabetes and lowered serum testosterone concentrations evaluated the change in hepatic steatosis as measured by liver fat fraction on magnetic resonance imaging(MRI).RESULTS Of 88 patients enrolled in the index study,39 had liver MRIs of whom 20 received testosterone therapy and 19 received placebo.All patients had>5%hepatic steatosis at baseline and 38 of 39 patients met diagnostic criteria for NAFLD.Median liver fat at baseline was 15.0%(IQR 11.5%-21.1%)in the testosterone and 18.4%(15.0%-28.9%)in the placebo group.Median ALT was 34units/L(26-38)in the testosterone and 32units/L(25-52)in the placebo group.At week 40,patients receiving testosterone had a median reduction in absolute liver fat of 3.5%(IQR 2.9%-6.4%)compared with an increase of 1.2%in the placebo arm(between-group difference 4.7%P<0.001).After controlling for baseline liver fat,testosterone therapy was associated with a relative reduction in liver fat of 38.3%(95%confidence interval 25.4%-49.0%,P<0.001).CONCLUSION Testosterone therapy was associated with a reduction in hepatic steatosis in men with diabetes and low serum testosterone.Future randomised studies of testosterone therapy in men with NAFLD focusing on liver-related endpoints are therefore justified.

Relationship between Physical Activity Level, Hepatic Steatosis Presence, Metabolic Syndrome and the Risk of Developing Type 2 Diabetes in Men

Cardiovascular disease, cancer, respiratory and metabolic disease represent 63% of all deaths worldwide and are considered the major causes of morbidity and mortality. Physical inactivity is considered a case of public health, and other behavioral and metabolic risk factors, according to WHO (2011), such as smoking, increased blood pressure, increased blood glucose, hypercholesterolemia, overweight and obesity. The Nonalcoholic Fatty Liver Disease (NAFLD) is the most prevalent liver disease in adults, and can progress and be characterized as hepatic steatosis (HS) which is derived from the accumulation of lipids in hepatocytes, and histopathologic condition is more than 5% of the weight of liver. So, the purpose of this study is to identify the relationship between the physical activity level and the hepatic steatosis presence, metabolic syndrome and the risk of developing type 2 diabetes mellitus in men. We evaluated retrospectively medical records of 1399 men (40.7 ± 8.18 years old) who participated in the protocol of Preventive Health Check-up at Hospital Israelita Albert Einstein from January to October 2011. According to the results, it is concluded that there is a positive association between low physical activity level and the presence of Hepatic steatosis. The results further demonstrate that, despite the high BMI, blood levels of the subjects remained unchanged. Even without a positive association between these variables, the results showed a high risk behavior for the development of diabetes mellitus type 2.

Hepatic DDAH1 mitigates hepatic steatosis and insulin resistance in obese mice: Involvement of reduced S100A11 expression

Dimethylarginine dimethylaminohydrolase 1(DDAH1)is an important regulator of plasma asymmetric dimethylarginine(ADMA)levels,which are associated with insulin resistance in patients with nonalcoholic fatty liver disease(NAFLD).To elucidate the role of hepatic DDAH1 in the pathogenesis of NAFLD,we used hepatocyte-specific Ddah1-knockout mice(Ddah1HKO)to examine the progress of high-fat diet(HFD)-induced NAFLD.Compared to diet-matched flox/flox littermates(Ddah1f/f),Ddah1HKO mice exhibited higher serum ADMA levels.After HFD feeding for 16 weeks,Ddah1HKO mice developed more severe liver steatosis and worse insulin resistance than Ddah1f/f mice.On the contrary,overexpression of DDAH1 attenuated the NAFLD-like phenotype in HFD-fed mice and ob/ob mice.RNA-seq analysis showed that DDAH1 affects NF-kB signaling,lipid metabolic processes,and immune system processes in fatty livers.Furthermore,DDAH1 reduces S100 calcium-binding protein A11(S100A11)possibly via NF-kB,JNK and oxidative stress-dependent manner in fatty livers.Knockdown of hepatic S100a11 by an AAV8-shS100a11 vector alleviated hepatic steatosis and insulin resistance in HFD-fed Ddah1HKO mice.In summary,our results suggested that the liver DDAH1/S100A11 axis has a marked effect on liver lipid metabolism in obese mice.Strategies to increase liver DDAH1 activity or decrease S100A11 expression could be a valuable approach for NAFLD therapy.

An early screening model of pulse detection technology for hepatic steatosis

Background The increasing prevalence of hepatic steatosis presents a considerable challenge to public health.There is a critical need for the development of novel preventive and screening strategies for this condition.Thisstudy evaluated the potential applications of wrist pulse detection technology for the early detection of liverdiseases.The pulse time-domain features of a medical exam population with and without hepatic steatosis wereassessed to develop a screening model for this disease.Methods Participants were consecutively recruited from March 2021 to March 2022 in the medical examinationcenters of the Yueyang Hospital of Integrated Traditional Chinese and Western Medicine and the Shanghai Municipal Hospital of Traditional Chinese Medicine.Clinical data from 255 participants,including general information(sex,age,and body mass index),and data related to glucose and blood lipids(fasting plasma glucose,triglyceride,total cholesterol,high-density lipoprotein,and low-density lipoprotein levels)were collected.Wrist pulse signalswere acquired using a pulse detection device,and the pulse time-domain features,including t_(1),t_(4),t_(5),T,w_(1),w_(2),h_(2)/h_(1),h_(3)/h_(1),and h5/h_(1) were extracted.Participants were assigned to hepatic steatosis and non-hepatic steatosisgroups according to their abdominal ultrasound examination results.Their clinical data and pulse time-domainfeatures were compared using chi-square and parametric or non-parametric statistical methods.Three datasetswere used to construct screening models for hepatic steatosis based on the random forest algorithm.The datasetsfor modeling were defined as Dataset 1,containing blood glucose and lipid data and general information;Dataset2,containing time-domain features and general information;Dataset 3,containing time-domain features,bloodglucose and lipid data,and general information.The evaluation metrics,accuracy,precision,recall,F1-score,andareas under the receiver operating characteristic curve(AUC)were compared for each model.Results The time-domain features of the two groups differed significantly.The t_(1),t_(4),t_(5),T,h_(2)/h_(1),h_(3)/h_(1),w_(1),and w_(2) features were higher in the hepatic steatosis group than in the non-hepatic steatosis group(P<0.05),while the h5/h_(1) features were lower in the hepatic steatosis group than in the non-hepatic steatosis group(P<0.05).The screening models for hepatic steatosis based on both time-domain features and blood glucose andlipid data outperformed those based on time-domain features or blood markers alone.The accuracy,precision,recall,F1-score,and AUC of the combined model were 81.18%,80.56%,76.32%,79%,and 87.79%,respectively.These proportions were 1.57%,1.86%,1.76%,2%,and 3.54%higher,respectively,than those of the model basedon time-domain features alone and 3.14%,4.2%,2.64%,4%,and 6.47%higher,respectively,than those of themodel based on blood glucose and lipid alone.Conclusion The early screening model for hepatic steatosis using datasets that included pulse time-domainfeatures achieved better performance.The findings suggest that pulse detection technology could be used toinform the development of a mobile medical device or remote home monitoring system to test for hepatitissteatosis.

Activation of NLRP3 Inflammasome via Drp1 Overexpression in Kupffer Cells Aggravates Ischemia-reperfusion Injury in Hepatic Steatosis

Background and Aims:Donors with fatty livers are considered to address the shortage of livers for transplantation,but those livers are particularly sensitive to ischemia-reperfusion injury(IRI),and an increased incidence of graft failure is observed.Kupffer cells account for 20–35%of liver nonparenchymal cells,and have been shown to participate in the process of IRI and inflammatory reactions of hepatic steatosis.NOD-like receptor thermal protein domain-associated protein 3(NLRP3)is an intracellular sensor activated by Kupffer cells to promote generation and participates in IRI.Dynamics-associated protein 1(Drp1)is one of the main proteins regulating mitochondrial division and exacerbates IRI by affecting mitochondrial dynamics.The mechanism of interaction of Kupffer cells with Drp1 and NLRP3 to aggravate IRI has not been clarified.Methods:A mouse model of hepatic steatosis was established by feeding the mice with a high-fat diet.In vitro experiments were performed using AML12 normal mouse liver cells and RAW264.7 mononuclear macrophage cells cultured in medium with palmitate and oleic acid.Western blotting and immunohistochemical(IHC)staining were used to detect the expression of NLRPP3 and Drp1 in IRI in the control and high-fat diet groups.The expression of F4/80+cells during IRI in hepatic steatosis was verified by IHC staining,and the role of NLRPP3 and Drp1 in Kupffer-cell mediated IRI was investigated by targeting Drp-1 inhibition.Results:Drp1 and NLRP3 expression was increased during IRI in hepatic steatosis,and the expression of Drp1 and NLRP3 were decreased after the elimination of Kupffer cells.That indicated Kupffer cells were involved in the process of IRI in hepatic steatosis through the action of Drp1 and NLRP3.After Drp1 inhibition,liver function was restored and NLRP3 expression level was reduced.Conclusions:Kupffer cells aggravated IRI in hepatic steatosis via NLRP3 and Drp1.Drp1 inhibitors might be useful as specific therapeutics to alleviate IRI in hepatic steatosis and may have promise in case of liver donor shortage.

NF-κB/HDAC1/SREBP1c pathway mediates the inflammation signal in progression of hepatic steatosis

The transcription factor nuclear factor kappa B(NF-κB)is activated in hepatoctes in the pathogenesis of hepatic steatosis.However,the action mechanism of NF-κB remains to be established in the hepatic steatosis.In this study,the P50 subunit of NF-κB was found to promote the hepatic steatosis through regulation of histone deacetylase 1(HDAC1)in hepatocytes.The activity was supported by the phenotypes of P50 knockout(P50-KO)mice and P65 knockout(P65-KO)mice.Hepatic steatosis was reduced in the P50-KO mice,but not in the P65-KO mice.The reduction was a result of inhibition of HDAC1 activity in the P50-KO cells.Knockdown of Hdac1 gene led to suppression of hepatocyte steatosis in HepG2 cells.A decrease in sterol-regulatory element binding protein lc(SREBP1 c)protein was observed in the liver of P50-KO mice and in cell with Hdac1 knockdown.The decrease was associated with an increase in succinylation of SREBP1 c protein.The study suggests that P50 stabilizes HDAC1 to support the SREBP1 c activity in hepatic steatosis in the pathophysiological condition.Interruption of this novel pathway in the P50-KO,but not the P65-KO mice,may account for the difference in hepatic phenotypes in the two lines of transgenic mice.

Mesencephalic astrocyte-derived neurotrophic factor ameliorates steatosis in HepG2 cells by regulating hepatic lipid metabolism

BACKGROUND Nonalcoholic fatty liver disease (NAFLD) is a global metabolism-associated liver disease.Mesencephalic astrocyte-derived neurotrophic factor (MANF) is a newly discovered secreted protein that is involved in metabolic homeostasis.However,much remains to be discovered about its function in hepatic lipid metabolism;thus,we assessed whether MANF could regulate hepatic metabolism.AIM To establish in vivo and in vitro NAFLD models to explore the role of MANF in hepatic lipid metabolism.METHODS HepG2 cells treated with free fatty acids (FFAs) and ob/ob mice were used as NAFLD models.Liver tissues collected from wild type and ob/ob mice were used to detect MANF expression.Cells were treated with FFAs for different durations.Moreover,we used lentiviral constructs to establish overexpression and knockdown cell models in order to interfere with MANF expression levels and observe whether MANF influences hepatic steatosis.Western blot analysis and quantitative real-time PCR were used to detect protein and gene expression,and oil red O staining was used to visualize intracellular lipid droplets.RESULTS Hepatic MANF protein and mRNA expression in wild type mice were 10-fold and 2-fold higher,respectively,than those in ob/ob mice.The MANF protein was temporarily increased by 1.3-fold after stimulation with FFAs for 24 h and gradually decreased to 0.66-fold that of the control at the 72 h time point in HepG2 cells.MANF deficiency upregulated the expression of genes involved infatty acid synthesis,cholesterol synthesis,and fatty acid uptake and aggravated HepG2 cell steatosis,while MANF overexpression inhibited fatty acid synthesis and uptake and cholesterol synthesis,and rescued HepG2 cells from FFAsinduced steatosis.Furthermore,a significant decrease in triglyceride levels was observed in the MANF overexpression group compared with the control group(0.4288±0.0081 mmol/g vs 0.3746±0.0121 mmol/g,P <0.05) upon FF As treatment.There was also a 17%decrease in intracellular total cholesterol levels between the MANF overexpression group and the control group (0.1301±0.0059mmol/g vs 0.1088±0.0009 mmol/g,P <0.05) upon FF As treatment.Moreover,MANF suppressed lipid deposition in HepG2 cells.CONCLUSION Our findings indicate that MANF improves the phenotype of liver cell steatosis and may be a potential therapeutic target in hepatic steatosis processes.

Loss of hnRNP A1 in murine skeletal muscle exacerbates high-fat diet-induced onset of insulin resistance and hepatic steatosis

Impairment of glucose(Glu)uptake and storage by skeletal muscle is a prime risk factor for the development of metabolic diseases.Heterogeneous nuclear ribonucleoprotein A1(hnRNP Al)is a highly abundant RNA-binding protein that has been implicated in diverse cellular functions.The aim of this study was to investigate the function of hnRNP A1 on muscle tissue insulin sensitivity and systemic Glu homeostasis.Our results showed that conditional deletion of hnRNP Al in the muscle gave rise to a severe insulin resistance phenotype in mice fed a high-fat diet(HFD).Conditional knockout mice fed a HFD showed exacerbated obesity,insulin resistance,and hepatic steatosis.In vitro interference of hnRNP Al in C2C12 myotubes impaired insulin signal transduction and inhibited Glu uptake,whereas hnRNP Al overexpression in C2C12 myotubes protected against insulin resistance induced by supraphysiological concentrations of insulin.The expression and stability of glycogen synthase(gysl)mRNA were also decreased in the absence of hnRNP A l.Mechanistically,hnRNP Al interacted with gys l and stabilized its mRNA,thereby promoting glycogen synthesis and maintaining the insulin sensitivity in muscle tissue.Taken together,our findings are the first to show that reduced expression of hnRNP Al in skeletal muscle affects the metabolic properties and systemic insulin sensitivity by inhibiting glycogen synthesis.

Adoptive transfer of Pfkfb3-disrupted hematopoietic cells to wild-type mice exacerbates diet-induced hepatic steatosis and inflammation

Background and objectives:Hepatic steatosis and inflammation are key characteristics of non-alcoholic fatty liver disease(NAFLD).However,whether and how hepatic steatosis and liver inflammation are differentially regulated remains to be elucidated.Considering that disruption of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(Pfkfb3/iPfk2)dissociates fat deposition and inflammation,the present study examined a role for Pfkfb3/iPfk2 in hematopoietic cells in regulating hepatic steatosis and inflammation in mice.Methods:Pfkfb3-disrupted(Pfkfb3^(+/-))mice and wild-type(WT)littermates were fed a high-fat diet(HFD)and examined for NAFLD phenotype.Also,bone marrow cells isolated from Pfkfb3^(+/-)mice andWT mice were differentiated into macrophages for analysis of macrophage activation status and for bone marrow transplantation(BMT)to generate chimeric(WT/BMT-Pfkfb3^(+/-))mice in which Pfkfb3 was disrupted only in hematopoietic cells and control chimeric(WT/BMT-WT)mice.The latter were also fed an HFD and examined for NAFLD phenotype.In vitro,hepatocytes were co-cultured with bone marrowderived macrophages and examined for hepatocyte fat deposition and proinflammatory responses.Results:After the feeding period,HFD-fed Pfkfb3^(+/-)mice displayed increased severity of liver inflammation in the absence of hepatic steatosis compared with HFD-fed WT mice.When inflammatory activation was analyzed,Pfkfb3^(+/-)macrophages revealed increased proinflammatory activation and decreased anti-proinflammatory activation.When NAFLD phenotype was analyzed in the chimeric mice,WT/BMT-Pfkfb3^(+/-) mice displayed increases in the severity of HFD-induced hepatic steatosis and inflammation compared with WT/BMT-WT mice.At the cellular level,hepatocytes co-cultured with Pfkfb3^(+/-) macrophages revealed increased fat deposition and proinflammatory responses compared with hepatocytes co-cultured with WT macrophages.Conclusions:Pfkfb3 disruption only in hematopoietic cells exacerbates HFD-induced hepatic steatosis and inflammation whereas the Pfkfb3/iPfk2 in nonhematopoietic cells appeared to be needed for HFD feeding to induce hepatic steatosis.As such,the Pfkfb3/iPfk2 plays a unique role in regulating NAFLD pathophysiology.