Hepatic Infiltration Research Articles

Potential hepatoprotective effects of Cistanche deserticola Y.C. Ma: Integrated phytochemical analysis using UPLC-Q-TOF-MS/MS, target network analysis, and experimental assessment.

Cistanche deserticola Y.C. Ma (CD) possesses hepatoprotective activity, while the active ingredients and involved mechanisms have not been fully explored. The objective of this study was to investigate the chemical composition and hepatoprotective mechanisms of CD. We primarily used ultra-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) to identify the phenylethanoid glycoside (PhG) components of CD. Then, network analysis was used to correlate and predict the pharmacology of the identified active components of PhGs with hepatoprotection. Next, the mechanisms of the core components and targets of action were explored by cellular assays and toll-like receptor 4 (TLR4) target competition assays. Finally, its hepatoprotective effects were further validated in in vivo experiments. The results showed that a total of 34 PhGs were identified based on the UPLC-Q-TOF-MS/MS method. Echinacoside (ECH) was identified as the key ingredient, and TLR4 and nuclear factor-kappa B (NF-κB) were speculated as the core targets of the hepatoprotective effect of CD via network analysis. The cellular assays confirmed that PhGs had significant anti-inflammatory activity. In addition, the real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot indicated that ECH notably reduced the levels of interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α), as well as the mRNA expression of TLR4, TNF-α, and IL-6, and decreased the high expression of the TLR4 protein, which in turn downregulated the myeloid differentiation factor 88 (MyD88), p-P65 and TNF-α proteins in the inflammatory model. The target competition experiments suggested that ECH and LPS could competitively bind to the TLR4 receptor, thereby reducing the expression of TLR4 downstream proteins. The results of in vivo studies showed that ECH significantly ameliorated LPS-induced hepatic inflammatory infiltration and liver tissue damage and reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in mice. Moreover, ECH remarkably inhibited the release of inflammatory factors such as TNF-α, IL-6, IL-1β, and MCP-1 in the serum of mice, exerting the hepatoprotective effect by the TLR4/NF-κB signaling pathway. More importantly, ECH could act as a potential inhibitor of TLR4 and deserves further in-depth study. Our results could provide a basis for exploring the hepatoprotective properties of CD.

The Preliminary Results for Evaluating Cocoa Butter's Hepatoprotective Effects against Lipid Accumulation and Inflammation in Adult Male Rats Chronically Fed Ethanol.

The purpose of this study was to clarify the role of saturated fats from cocoa butter (plant source) compared with lard (animal source) on alcoholic liver damage in rats. Male Wistar rats were fed either a control diet (C) or an ethanol diet (E), and the dietary fats (corn oil, olive oil, and safflower oil) of these two diets were further replaced by lard (CL, EL) or cocoa butter (CC, EC). After 8-week feeding, plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, hepatic triglyceride (TG) levels, plasma intercellular adhesion molecular (ICAM)-1 levels, hepatic cytochrome P450 2E1 (CYP2E1) protein expression, and hepatic interleukin (IL)-1β significantly increased in the E group compared to the C group. In addition, hepatic histopathological scores of fatty changes, inflammatory cell infiltration, and degeneration and necrosis in the E group were significantly higher compared to those in the C group. However, fatty changes were significantly inhibited only in the EC group as well as hepatic inflammatory cell infiltration, degeneration, and necrosis being significantly lower in the EL and EC groups. Plasma ICAM-1 and hepatic tumor necrosis factor (TNF)-α, IL-1β, IL-6, and IL-10 levels were significantly lower in the EL and EC groups than those in the E group. Moreover, a correlation analysis showed that hepatic histopathological scores of degeneration and necrosis were significantly positively correlated with erythrocytic oleic acid (C18:1) and were negatively correlated with linoleic acid (C18:2). In conclusion, cocoa butter protected the liver against lipid accumulation and inflammation in rats chronically fed ethanol.

NLRP3 activation in neutrophils induces lethal autoinflammation, liver inflammation, and fibrosis.

Sterile inflammation is a central element in liver diseases. The immune response following injurious stimuli involves hepatic infiltration of neutrophils and monocytes. Neutrophils are major effectors of liver inflammation, rapidly recruited to sites of inflammation, and can augment the recruitment of other leukocytes. The NLRP3 inflammasome has been increasingly implicated in severe liver inflammation, fibrosis, and cell death. In this study, the role of NLRP3 activation in neutrophils during liver inflammation and fibrosis was investigated. Mouse models with neutrophil-specific expression of mutant NLRP3 were developed. Mutant mice develop severe liver inflammation and lethal autoinflammation phenocopying mice with a systemic expression of mutant NLRP3. NLRP3 activation in neutrophils leads to a pro-inflammatory cytokine and chemokine profile in the liver, infiltration by neutrophils and macrophages, and an increase in cell death. Furthermore, mutant mice develop liver fibrosis associated with increased expression of pro-fibrogenic genes. Taken together, the present work demonstrates how neutrophils, driven by the NLRP3 inflammasome, coordinate other inflammatory myeloid cells in the liver, and propagate the inflammatory response in the context of inflammation-driven fibrosis.

S3296 Acute on Chronic Liver Disease Due to Hepatic Infiltration of Chronic Lymphoproliferative Disorder of Natural Killer Cells

Introduction: We report a case of acute on chronic liver disease with a predominantly cholestatic pattern which was diagnosed as hepatic infiltration of chronic lymphoproliferative disorder of natural killer cells (CLPD-NK) after extensive workup and an initial false-negative liver biopsy. Case Description/Methods: A 54-year-old woman with alcohol use disorder, celiac disease, and Hashimoto thyroiditis initially presented with 8-week history of night sweats, fever and jaundice. She reported taking daily naproxen for a few weeks. The physical exam was remarkable for jaundice. Labs showed bilirubin of 2.5, AST 869, ALT 543, alkaline phosphatase 966 and INR with 0.9. Right upper quadrant ultrasound showed no ductal dilation with a non-specific hypoechoic area in the superior right lobe and nodular appearance of the liver. Autoimmune workup revealed positive anti-smooth muscle antibody in a low titer but was otherwise negative. A liver biopsy showed massive hepatic necrosis with marked pan-acinar lymphocytic inflammation. The leading cause of her liver injury was thought to be drug-induced due to naproxen in a background of chronic liver disease. Her liver disease progressed, developing ascites, encephalopathy and pruritus refractory to treatment. Nine months after initial presentation she presented with a hemoglobin of 7, MCV 104, LDH 590, Haptoglobin < 10, total bilirubin 2.9, and ferritin of 4289. WBC count was noted to have an absolute increase in lymphocytes, bone marrow biopsy showed CLPD-NK. A second liver biopsy was performed showing diffuse lymphohistiocytic infiltrate and flow cytometry revealed atypical lymphocytes/NK cell population. The patient was started on chemotherapy which led to clinical and biochemical improvement of her liver disease. (Figure) Discussion: CLPD-NK is a rare and heterogeneous disorder. Unlike other NK cell disorders, CLPD-NKs is not associated with EBV infection. This disorder follows an indolent course, making it difficult to detect it in the initial stages. Hepatic abnormalities in patients with lymphoproliferative disorders can occur from direct infiltration by abnormal cells, bile duct obstruction by lymphadenopathy, paraneoplastic syndrome, hemophagocytic syndrome, and reactivation of viral hepatitis. Our patient is to our knowledge the first case reported of hepatic infiltration from CLPD-NK. Treatment is indicated in symptomatic patients. Hence, early diagnosis is essential so patients can receive timely administration of immunosuppressive therapy.Figure 1.: Liver with diffuse lymphohistiocytic infiltrate. Sections show an atypical lymphohistiocytic infiltrate of the liver and composed of small to medium-sized cells that stain CD2+, CD3+, CD4 and CD8 with down-regulation, CD3, CD5+, CD7+, CD20 focally+, CD68+, CD163+, and Granzyme B +.

S3613 Gastric Spindle Cell Leiomyosarcoma Is Associated With Chronic Helicobacter pylori Infection

Introduction: Chronic Helicobacter pylori (HP) infection has been shown to be strongly associated with development of gastric malignancies, mostly gastric adenocarcinomas and lymphomas. We present a case of primary gastric leiomyosarcoma in a patient with persistent epigastric pain and HP infection. Case Description/Methods: A 59-year-old female presented to clinic with persistent, postprandial epigastric pain. Cardiopulmonary workup was negative. She had a history of esophageal leiomyoma resected endoscopically with negative margins 10 years ago. She was treated in the past for HP infection but never had repeat testing to confirm eradication. Abdominal computed tomography showed focal hepatic infiltration. She was advised to have a repeat upper endoscopy for surveillance but was lost to follow-up due to psychiatric comorbidities. She presented 6 years later with epigastric pain. Repeat CT abdomen showed a new 2.8 cm hypoattenuating lesion in the lesser curvature of the stomach. Patient did not follow up due to psychosocial issues. Two years later, she returned to clinic for epigastric pain and underwent repeat upper endoscopy and endoscopic ultrasound (EUS) with fine needle aspiration (FNA). EGD showed sub-gastric nodule attached gastric wall, about 30 mm in maximal dimension and gastric biopsy confirmed active HP infection. The EUS showed a 31.7x23.5 mm hypoechoic homogeneous gastric mass along the lesser gastric curvature. Cytology showed cellular spindle cell neoplasia with myogenic differentiation concerning for possible leiomyoma or leiomyosarcoma. Abdominal magnetic resonance imaging and chest computed tomography revealed no evidence of abdominal or thoracic metastasis. Subsequently, partial underwent sleeve gastrectomy. Pathology revealed Grade 1 myxoid leiomyosarcoma with negative margins. (Figure) Discussion: Prior studies demonstrated associations between HP infection and gastric malignancies - typically gastric adenocarcinoma or lymphomas. Few studies investigated the relationship between HP infection and gastric leiomyosarcomas.4,5 Importantly, this patient never had clearance of HP infection and may have allowed a leiomyosarcoma to develop. It is imperative that clinicians confirm eradication of HP infection given risk of leiomyosarcoma development in addition to other malignancies HP infections are known to cause.Figure 1.: Endoscopic and histologic examination of gastric leiomyosarcoma. (A) Endoscopic ultrasound visualization demonstrating tumor within lesser curvature of stomach. (B) Upper endoscopy revealing tumor approximately 4 cm from gastroesophageal junction. (C) Hematoxylin and eosin (H&E) stain of gastric tumor biopsy demonstrating spindle cell proliferation in a myxoid background with significant nuclear atypia and pleomorphism consistent with myxoid leiomyosarcoma. Magnification x400. (D) Immunohistochemical staining of gastric lesion biopsy demonstrating strong immunoreactivity to desmin. Not shown are positive stains for smooth muscle actin (SMA) as well as negative stains for Cytokeratin AE1/AE3, Myogenin, MyoD1, DOG-1, CD117, S100, SOX10, CD34, ALK-1 and ER. Positive and negative controls stained appropriately but are not included.

S2987 Esophageal Varices in Myelofibrosis Associated Non-Cirrhotic Portal Hypertension: A Call to Screening

Introduction: Myelofibrosis is characterized by bone marrow hematopoetic failure, leading to extramedullary hematopoiesis and can be secondary to various causes such as primary myelofibrosis or chronic proliferative neoplasms i.e. polycythemia vera myelofibrosis and essential thrombocythemia.1 Mylefibrosis in the setting of primary polycythemia can led to non-cirrhotic portal hypertension with esophageal varices as demonstrated in this clinical presentation. Although esophageal varices are a relatively uncommon complication in this setting given its increased mortality, some have recommended these patients undergo screening and surveillance upper endoscopy. Case Description/Methods: A 62 year old man presented to emergency department with hematemesis. Past medical history included secondary myelofibrosis from polycythemia vera on daily aspirin and ruxolitinib. Physical examination was pertinent for splenomegaly and hepatomegaly without ascites. Upper endoscopy (EGD) revealed diffuse gastric erosions and small esophageal varices with no high risk stigmata. Initial laboratory evaluation revealed WBC of 2.8 x 103/mcL, hemoglobin 7.0 g/dL, platelets 250 x 103/mcL, and normal liver associated enzymes. An MRI of his liver showed massive splenomegaly up to 27.4 cm, hepatomegaly up to 27.6 cm, esophageal varices, and an otherwise normal appearing liver. He had an unremarkable abdominal doppler ultrasound evaluation with normal hepatic and hepatopetal flow in the portal venous system. Fibroscan was obtained with kPa of 12. Chronic liver disease workup including infectious, autoimmune, and metabolic causes was unremarkable. The patient declined liver biopsy and the plan was made for follow-up EGD in 1 year to survey the small varices seen on the original EGD. Discussion: While the exact mechanism of portal hypertension secondary to myelofibrosis is still under investigation, several pathophysiologic mechanisms have been proposed including: hepatic infiltration of hematopoietic stem cells, sinusoidal fibrosis, increased pre-hepatic flow as a result of profound splenomegaly, and portal venous insults.2 Portal hypertension has been observed in 10% to 17% of patients with myelofibrosis.3 Given the increased risk of developing esophageal varices, patients in this population may benefit from variceal screening to proactively treat and prevent variceal hemorrhage. The role of fibroscan in risk stratifying these patients remains to be established with most studies limited to small retrospective case series.

Oxidative Stress Biomarkers Activities and Histological Changes in African Catfish (Clarias gariepinus) Exposed to Carwash Wastewater

The study evaluated the acute toxicity of vehicle wastewater on African catfish (Clarias gariepinus). With a weight range of 250–275.5g and a mean length of 23.5-28.5cm, 150 post-juvenile catfish were purchased from Dalar Kifi Fish Farm BUK New site road, Kano State, and acclimated in a tank for 14 days. Fish were exposed to different vehicle wastewater concentrations of 0.0, 20, 40, 60, and 80% v/v with 5-level exposure concentrations during the acute test (96 hours) in a Completely Randomized Design (CRD) employing renewal bioassay. The acute bioassay showed that the LC50 for the wastewater was 56 percent by volume at 96 hours. Compared to the control, the exposed fish exhibited abnormal swimming, erratic opercular movement, reflex loss, mucus secretion, increased air gulping with an increase in wastewater concentration. Except for water temperature, Biochemical Oxygen Demand (BOD) and pH, the remaining physicochemical parameters were not within the WHO/FAO recommended Standard The order of concentrations of the heavy metal in the wastewater was Cu &gt; Pb &gt; Cd &gt; Cr &gt; Ni. Antioxidant biomarker activities showed that Glutathione reductase (GSH) reduced in the same manner while Catalase (CAT) and Superoxide dismutase (SOD) activities rose considerably (P&lt;0.05) in the liver and gill tissues. Histopathological changes observed in gill tissues were as follows: Hyperplasia, necrosis, primary and secondary lamellae distortion, and epithelial thickening distortion. Hepatic cell breakdown, fatty degeneration, cellular infiltration, localized fibrosis, and cellular necrosis were all visible in the liver tissues. It can be inferred that changes in the histology and activity of the oxidative stress enzymes in fish tissues after exposure to wastewater harmed the experimental fish. As a result of their effects on fish and other non-target aquatic biotas, it is advised that the competent authorities create initiatives to reduce the indiscriminate discharge of untreated vehicle wash wastewater.

Chronic Lymphocytic Leukemia Involving the Liver Without High-Grade Transformation

Chronic lymphocytic leukemia is most frequently encountered among the elderly population. Hepatic infiltration may be present at the time of diagnosis, but significant extranodal involvement rarely occurs without high-grade transformation. Abnormal liver enzymes have important prognostic implications in terms of staging the disease and initiating treatment. We describe an atypical presentation of chronic lymphocytic leukemia in a patient who presented with rising liver enzyme levels, extensive hepatic infiltration, and portal hypertension but with no evidence of high-grade transformation on liver biopsy.

Metformin keeps CD8+ T cells active and moving in NASH-HCC immunotherapy

Metformin keeps CD8+ T cells active and moving in NASH-HCC immunotherapy

Effect of Chlorogenic Acid via Upregulating Resolvin D1 Inhibiting the NF-κB Pathway on Chronic Restraint Stress-Induced Liver Inflammation.

Chronic stress can cause chronic inflammatory injury to the liver. Chlorogenic acid (CGA) is known to have a wide range of biological activities and anti-inflammatory effects. Resolvin D1 (RvD1) is a polyunsaturated fatty acid derivative that has inhibitory effects on a variety of inflammatory diseases. However, whether CGA can inhibit liver inflammation in chronic stress through RvD1 remains unclear. In this work, male rats were subjected to restraint stress for 6 h every day and built a chronic stress model for 21 days. CGA (100 mg/kg) was administered intragastrically 1 h before restraint, with intraperitoneal injection of RvD1 inhibitor WRW4 (antagonist of FPR2, 0.1 mg/kg) or WRW4 solution every 2 days for 30 min before CGA administration. CGA reduced hepatic hemorrhage and inflammatory cell infiltration, alleviated hepatic injury, decreased the activation of the NF-κB pathway and the expression of interleukin 1β, interleukin 6, and tumor necrosis factor α in the liver, and increased RvD1 in the serum and liver. The therapeutic effect of CGA was blocked after WRW4 intervention. These results suggest that the protective effects of CGA mediate the NF-κB pathway by upregulating the generation of RvD1. Above all, this research demonstrates the liver protective effect of CGA and provides a potential treatment strategy for chronic inflammatory disease.

Ileal bile acid transporter inhibition in Cyp2c70 KO mice ameliorates cholestatic liver injury

Cyp2c70 is the liver enzyme in rodents responsible for synthesis of the primary 6-hydroxylated muricholate bile acid (BA) species. Cyp2c70 KO mice are devoid of protective, hydrophilic muricholic acids, leading to a more human-like BA composition and subsequent cholestatic liver injury. Pharmacological inhibition of the ileal BA transporter (IBAT) has been shown to be therapeutic in cholestatic models. Here, we aimed to determine if IBAT inhibition with SC-435 is protective in Cyp2c70 KO mice. As compared to WT mice, we found male and female Cyp2c70 KO mice exhibited increased levels of serum liver injury markers, and our evaluation of liver histology revealed increased hepatic inflammation, macrophage infiltration, and biliary cell proliferation. We demonstrate serum and histologic markers of liver damage were markedly reduced with SC-435 treatment. Additionally, we show hepatic gene expression in pathways related to immune cell activation and inflammation were significantly upregulated in Cyp2c70 KO mice and reduced to levels indistinguishable from WT with IBAT inhibition. In Cyp2c70 KO mice, the liver BA content was significantly increased, enriched in chenodeoxycholic acid, and more hydrophobic, exhibiting a hydrophobicity index value and red blood cell lysis properties similar to human liver BAs. Furthermore, we determined IBAT inhibition reduced the total hepatic BA levels but did not affect overall hydrophobicity of the liver BAs. These findings suggest that there may be a threshold in the liver for pathological accretion of hydrophobic BAs and reducing hepatic BA accumulation can be sufficient to alleviate liver injury, independent of BA pool hydrophobicity.

Gut microbiota–bile acid–intestinal Farnesoid X receptor signaling axis orchestrates cadmium-induced liver injury

Cadmium (Cd) is a widely prevalent environmental pollutant that accumulates in the liver and induces liver injury. The mechanism of Cd-induced liver injury remains elusive. Our study aimed to clarify the mechanism by which changes in the gut microbiota contribute to Cd-induced liver injury. Here, a murine model of liver injury induced by chronic Cd exposure was used. Liver injury was assessed by biochemistry and histopathology. Expression profiles of genes involved in bile acid (BA) homeostasis, inflammation and injury were assessed via Realtime-PCR and Western-blot. 16S rRNA gene sequencing and mass spectrometry-based metabolomics were used to investigate changes in the gut microbiota and its metabolites in the regulation of Cd-induced liver injury. Here, we showed that Cd exposure induced hepatic ductular proliferation, hepatocellular damage and inflammatory infiltration in mice. Cd exposure induced gut microbiota dysbiosis and reduced the fecal bile salt hydrolase activity leading to an increase of tauro-β-muricholic acid levels in the intestine. Cd exposure decreased intestine FXR/FGF-15 signaling and promoted hepatic BA synthesis. Furthermore, the mice receiving fecal microbiota transplantation from Cd-treated mice showed reduced intestinal FXR/FGF-15 signaling, increased hepatic BA synthesis, and liver injury. However, the depletion of the commensal microbiota by antibiotics failed to change these indices in Cd-treated mice. Finally, the administration of the intestine-restricted FXR agonist fexaramine attenuated the liver injury, improved the intestinal barrier, and decreased hepatic BA synthesis in the Cd-treated mice. Our study identified a new mechanism of Cd-induced liver injury. Cd-induced gut microbiota dysbiosis, decreased feces BSH activity, and increased intestinal T-βMCA levels led to an inhibition of intestinal FXR/FGF-15 signaling and an increase in hepatic BA synthesis, ultimately facilitating the development of hepatic ductular proliferation, inflammation, and injury in mice. This study expands our understanding of the health hazards caused by environmental Cd pollution.

Characterization and Quantification of Oxidized High Mobility Group Box 1 Proteoforms Secreted from Hepatocytes by Toxic Levels of Acetaminophen.

The high mobility group box 1 (HMGB1), which is released during acute acetaminophen (APAP) overdose, is thought to mediate a subsequent immune response, particularly hepatic infiltration of macrophages. The redox behavior of HMGB1 and the proteoforms of HMGB1 present in oxidative environments has been the subject of a number of confusing and contradictory studies. Therefore, a stable isotope dilution two-dimensional nanoultrahigh-performance liquid chromatography parallel reaction monitoring/high-resolution mass spectrometry method was developed in order to characterize and quantify oxidative modifications to the cysteine (Cys) residues (Cys-23, Cys-45, and Cys-106) that are present in HMGB1. Disulfide linkages were determined using carbamidoethyl derivatization before and after reduction as well as by direct analysis of disulfide cross-linked peptides. A stable isotope labeled form of HMGB1 was used as an internal standard to correct for sample to sample differences in immunoaffinity precipitation, derivatization, and electrospray ionization. Four discrete HMGB1 proteoforms were found to be released from a hepatocarcinoma cell model of APAP overdose after 24 h. Fully reduced HMGB1 with all three Cys-residues in their free thiol state accounted for 18% of the secreted HMGB1. The proteoform with disulfide between Cys-23 and Cys-45 accounted for 24% of the HMGB1. No evidence was obtained for a disulfide cross-link between Cys-106 and the other two Cys-residues. However, 45% of the HMGB1 formed a cross-link with unidentified intracellular proteins via an intermolecular disulfide bond, and 12% was present as the terminally oxidized cysteic acid. Surprisingly, there was no evidence for the formation of HMGB1 disulfides with GSH or other low molecular weight thiols. Secreted plasma HMGB1 Cys-23/Cys45 disulfide proteoform together with the Cys-106/protein disulfide proteoforms could potentially serve as early biomarkers of hepatoxicity after APAP overdose as well as biomarkers of drug-induced liver injury.

Integrated strategy of network pharmacology, molecular docking, HPLC-DAD and mice model for exploring active ingredients and pharmacological mechanisms of Penthorum chinense Pursh against alcoholic liver injury

Ethnopharmacological relevancePenthorum chinense Pursh (PCP, Saxifragaceae) is an edible plant and frequently-used Chinese herbal medicine, and is commonly used as Miao medicine in China. It showed well effect on alcoholic liver injury (ALI), but studies on its active ingredients and mechanisms against ALI remain at the starting stage. Aim of the studyThis work aims to explore the active ingredients and pharmacological mechanisms of PCP against ALI. Materials and methodsFirst, network pharmacology was applied to decipher the potential active ingredients and pharmacological mechanisms of PCP against ALI by ingredient identification, ADMET evaluation, target identification, network construction and analysis, protein-protein interaction (PPI) analysis, and gene enrichment analysis. Second, molecular docking was used to explore the interaction between key active ingredient and hub protein of PCP against ALI. Then, the ingredient analysis of PCP aqueous extract and semiquantitative analysis of key active ingredient were carried out on HPLC-DAD. Subsequently, mice with ALI were used to investigate the therapeutic effect or verify the predicted mechanisms of PCP or key active ingredient against ALI by analyzing body weight, liver index, ALT and AST activities in serum and liver tissues, oxidation related indices (SOD activity, GSH level and MDA level) in liver tissues, histopathology of liver tissues (oil red O, hematoxylin-eosin and DAB-TUNEL staining), and changes of related proteins (PI3K, Akt, p-Akt, Bax and Bcl-2) in liver tissues with the aid of Western blot. ResultsNetwork pharmacology showed that the active ingredients and related genes of PCP against ALI comprised 10 ingredients and 52 genes. Based on the result of ingredient analysis of PCP aqueous extract, quercitrin was identified as the key active ingredient of PCP against ALI. PPI analysis indicated that AKT1 was the hub gene of PCP against ALI, and molecular docking suggested that there were good interaction between quercetin and Akt1 protein. Gene enrichment analysis showed that the pivotal molecular mechanism of PCP against ALI might be to inhibit hepatocyte apoptosis via activation of PI3K-Akt signaling pathway. PCP and quercitrin showed anti-ALI effect by offsetting weight loss and increase of liver index, and reversing the imbalance of oxidative stress and histopathological changes of liver tissues (abnormal fatty acid metabolism, hepatic cord swelling and inflammatory cell infiltration) in mice with ALI. PCP caused the decrease of DAB-TUNEL-positive cells, upregulated the anti-apoptotic proteins (PI3K, Akt and p-Akt) levels and the ratio of p-Akt/Akt, and downregulated pro-apoptotic protein (Bax) level and the ratio of Bax/Bcl-2 in liver tissues of mice with ALI, indicating that the mechanism of PCP against ALI involved in inhibiting hepatocyte apoptosis via activation of PI3K-Akt signaling pathway. ConclusionPCP and quercitrin showed well anti-ALI effect. The key active ingredient of PCP against ALI was identified as quercitrin. The underlying pharmacological mechanisms of PCP against ALI may be related to PI3K-Akt signaling pathway-mediated inhibition of hepatocyte apoptosis. This work provided new evidence to support the application of PCP in treatment of ALI, and a research basis for the research and development of functional foods or drugs against ALI from PCP.

Liposome-encapsulated curcumin attenuates HMGB1-mediated hepatic inflammation and fibrosis in a murine model of Wilson's disease.

Wilson's disease (WD) is an inherited disorder of copper metabolism with predominant hepatic manifestations. Left untreated, it can be fatal. Current therapies focus on treating copper overload rather than targeting the pathophysiology of copper-induced liver injuries. We sought to investigate whether liposome-encapsulated curcumin (LEC) could attenuate the underlying pathophysiology of WD in a mouse model of WD. Subcutaneous administration in a WD mouse model with ATP7B knockout (Atp7b-/-) resulted in robust delivery of LEC to the liver as determined by in-vitro and in-vivo imaging. Treatment with LEC attenuated hepatic injuries, restored lipid metabolism and decreased hepatic inflammation and fibrosis, and thus hepatosplenomegaly in Atp7b-/- mice. Mechanistically, LEC decreased hepatic immune cell and macrophage infiltration and attenuated the hepatic up-regulation of p65 by preventing cellular translocation of high-mobility group box-1 (HMGB-1). Moreover, decreased translocation of HMGB1 was associated with reduced splenic CD11b+/CD43+/Ly6CHi inflammatory monocyte expansion and circulating level of proinflammatory cytokines. Nevertheless there was no change in expression of oxidative stress-related genes or significant copper chelation effect of LEC in Atp7b-/- mice. Our results indicate that treatment with subcutaneous LEC can attenuate copper-induced liver injury in an animal model of WD via suppression of HMGB1-mediated hepatic and systemic inflammation. These findings provide important proof-of-principle data to develop LEC as a novel therapy for WD as well as other inflammatory liver diseases.

Extracts and fractions of Gymnema sylvestre restore liver and kidney function by regulating tissue and serum transaminases in the type 2 diabetic rat

Diabetes mellitus usually induce kidney and liver complications, characterized by an increase in transaminases activity. The aim of this study was to evaluate the activity of Gymnema sylvestre on the variation of tissue and serum transaminases activities in high sucrose diet and dexamethasone-induced diabetic rats. The animals were fed with high sucrose diet and 20% sugar water for 12 weeks then, received an intraperitoneal injection of dexamethasone (8mg/kg) once a day for 5 days between 07 and 08 am. After induction, the insulin sensitivity test was performed and the animals were divided into 7 groups. The experimental groups received extracts (aqueous and methanolic) and fractions (methylene chloride and methanol) of G. sylvestre once a day for 14 days. At the end of experiment, the animals were sacrificed and the blood, liver and kidney were collected for evaluation of transaminases and histopathology reduced by the plant. The activity of liver and kidney transaminases of the diabetic negative control (DNC) group on days 1 and 14 had significantly elevated (P&lt;0.001) compared with this of the normoglycemic negative control (NNC) group and the different treated groups. This trend was also observed at the serum level but only with ASAT. Histological analysis revealed hepatic leukocyte infiltrations and mesangial expansions in the urinary space in the DNC group on days 1 and 14. These tissue abnormalities were corrected in groups receiving extracts and fractions of G. sylvestre. This study suggests that the extracts and fractions of G. sylvestre restore tissue damage in diabetic rats.

Chemotactic cytokines secreted from Kupffer cells contribute to the sex-dependent susceptibility to non-alcoholic fatty liver diseases in mice

AimsThe global prevalence of non-alcoholic fatty liver disease (NAFLD) has rapidly increased over the last decade due to an elevated occurrence of metabolic syndromes. Importantly, the prevalence and severity of NAFLD is higher in men than in women. Therefore, in the present study we endeavored to identify the mechanistic disparity between male and female mice. Main methodsGlobal gene transcriptomics analysis was done with the high-fat diet (HFD)-induced NAFLD model of male, female, and ovariectomized (OVX) female mice. The expression of CCL2, CXCL2, and CXCL10 in mRNA level and serum protein level was done by qPCR and ELISA each. Immunohistochemistry staining was used to observe hepatic immune cell infiltration. To analyzing portion of immune cells, flow cytometry was done with isolated liver cells from HFD-fed male and female mice. Primary mouse liver cells were isolated from male and female mice for in vitro studies. Key findingsWe identified sex differences in inflammatory chemokines, CCL2, CXCL2, and CXCL10, with the expression of these chemokines enhanced in male and OVX, but not in female, mice after HFD feeding. Resident Kupffer cells (KCs) were identified as the major source of production of CCL2, CXCL2, and CXCL10 in the mouse NAFLD model. Notably, KCs obtained from male mice expressed higher levels of chemokines than those from female mice, indicating that KCs may mediate the sex discrepancy in NAFLD progression. SignificanceOur findings offer new insights into the pathology of sex-specific differences in NAFLD, involving chemokines and KCs.

Adipose lipolysis is important for ethanol to induce fatty liver in the National Institute on Alcohol Abuse and Alcoholism murine model of chronic and binge ethanol feeding.

Alcohol-associated liver disease (ALD) pathologies include steatosis, inflammation, and injury, which may progress to fibrosis, cirrhosis, and cancer. The liver receives ~60% of fatty acids from adipose tissue triglyceride hydrolysis, but the role of this lipolytic pathway in ALD development has not been directly examined in any genetic animal models with selective inactivation of adipose lipolysis. Using adipose-specific comparative gene identification-58 (CGI-58) knockout (FAT-KO) mice, a model of impaired adipose lipolysis, we show that mice deficient in adipose lipolysis are almost completely protected against ethanol-induced hepatic steatosis and lipid peroxidation when subjected to the National Institute on Alcohol Abuse and Alcoholism chronic and binge ethanol feeding model. This is unlikely due to reduced lipid synthesis because this regimen of ethanol feeding down-regulated hepatic expression of lipogenic genes similarly in both genotypes. In the pair-fed group, FAT-KO relative to control mice displayed increased hepatocyte injury, neutrophil infiltration, and activation of the transcription factor signal transducer and activator of transcription 3 (STAT3) in the liver; and none of these were exacerbated by ethanol feeding. Activation of STAT3 is associated with a marked increase in hepatic leptin receptor mRNA expression and adipose inflammatory cell infiltration. Our findings establish a critical role of adipose lipolysis in driving hepatic steatosis and oxidative stress during ALD development.

Glycyrrhetinic acid regulates impaired macrophage autophagic flux in the treatment of non-alcoholic fatty liver disease.

Macrophages are involved in hepatocyte steatosis and necroinflammation and play an important role in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). Impaired autophagy function (decreased autophagy or blocked autophagic flow) leads to cell damage and death and promotes NAFLD progression. The experimental and clinical research of glycyrrhetinic acid (GA) in the treatment of NAFLD has gradually attracted attention with clear pharmacological activities such as immune regulation, antiviral, antitumor, antioxidant, liver protection, and anti-inflammatory. However, the effects of GA on the STAT3-HIF-1α pathway and autophagy in macrophages are still unclear, and its mechanism of action in the treatment of NAFLD remains to be further elucidated. We constructed a NAFLD mouse model through a high-fat and high-sugar diet to investigate the therapeutic effects of GA. The results showed that GA reduced weight, improved the pathological changes and hepatic lipid deposition of liver, and abnormally elevated the levels of serum biochemical (AST, ALT, TG, T-CHO, LDL-C, and HDL-C) and inflammatory indexes (IL-1β, IL-4, IL-6, MCP-1, and TNF-α) in NAFLD mice. Further examination revealed that GA ameliorates excessive hepatic macrophage infiltration and hepatocyte apoptosis. The results of the cell experiments further elaborated that GA modulated the PA-induced macrophage STAT3-HIF-1α pathway and ameliorated impaired autophagic flux (blockade of autophagosome–lysosome fusion) and overactivation of inflammation. Excessive hepatocyte apoptosis caused by the uncontrolled release of inflammatory cytokines was also suppressed by GA.ConclusionThis study demonstrated that GA could regulate the STAT3-HIF-1α pathway of macrophages, ameliorate the impaired autophagy flux, and reduce the excessive production of inflammatory cytokines to improve the excessive apoptosis of liver cells, thus playing a therapeutic role on NAFLD.

Value of contrast-enhanced ultrasound in diagnosis and differential diagnosis of polypoid lesions of gallbladder ≥ 1 cm

ObjectivesTo evaluate the usefulness of Contrast-enhanced ultrasound (CEUS) in the diagnosis and differential diagnosis of Polypoid lesions of gallbladder (PLGs) ≥ 1 cm.MethodsA prospective analysis was performed on 180 patients with PLGs ≥ 1 cm. 175 cases were confirmed by pathological diagnosis and the remaining were confirmed by other imaging findings. The characteristics of lesions on conventional Ultrasonography (US) and CEUS were recorded.ResultsSignificant differences were observed in enhancement patterns between benign and malignant PLGs during both arterial (P < 0.001) and venous phases (P < 0.001). The malignant lesions typically yielded a “fast-in and fast-out” enhancement pattern. There was no significant difference in Arrival time (AT) between malignant and benign PLGs. If we consider wash-out time ≤ 40 s as a diagnostic standard for malignant lesions, the sensitivity, specificity, and accuracy were 88.24%, 85.62%, and 86.11%, respectively. Destruction of the Gallbladder (GB) wall was a particularly important indication of malignant PLGs, and the sensitivity, specificity, and accuracy were 93.33%, 92.12%, and 92.22%, respectively. The accuracy of CEUS in the diagnosis of PLGs, as well as malignant and benign lesions, was 92.22%, 92.47%, and 91.17%, respectively.ConclusionsThe “fast-in and fast-out” enhancement pattern, hyper-enhancement in comparison to the GB wall in the arterial phase, wash-out time ≤ 40 s, GB wall destruction, and hepatic parenchymal infiltration are the characteristic findings of malignant PLGs. Besides, CEUS provides a valuable reference to classify some of the benign lesions.