Hepatic Infiltration Research Articles

Replacement of fishmeal with processed canola meal in diets for juvenile Nile tilapia (Oreochromis niloticus): Growth performance, mucosal innate immunity, hepatic oxidative status, liver and intestine histology

The present study was carried out to evaluate the effect of processed canola meal (PCM) as fishmeal (FM) substitute in juvenile Nile tilapia diet. Firstly, canola meal (CM) was dephytinized using 2000 IU/kg phytase and subjected to methanolic ammonia solution (MAS). Finally, five isonitrogenous and isoenergetic diets were formulated by inclusion of 0 (control), 12.5, 25, 37.5 and 50% PCM at the expense of dietary FM. Each experimental diet was fed to three randomly allotted groups of 17 juvenile Nile tilapia with an average body weight of 3.5 ± 0.1 g for 36 days. Fish were feed three times a day until apparent satiation. Results showed that there were no statistically significant differences regarding SGR, final body weight and WG between control group and PCM2 (i.e., diet with 25% PCM inclusion level) (p > .05); however receiving diets with higher PCM content (>25%) resulted in significantly decreased growth indices (p < .05). Meanwhile, FCR, FI and apparent digestibility coefficient of protein (ADCprotein) did not differ among experimental groups (p > .05). Digestive enzyme activities, mucosal innate immunity, liver antioxidant enzyme activities and liver tissue malondialdehyde content (MDA) were not significantly affected by dietary inclusion of PCM (p > .05). However, relative gene expression of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) were significantly increased by dietary PCM content increment (p < .05). In line with growth performance, histological observations revealed that those fish received higher levels of dietary PCM content (i.e. 37.5 and 50%) showed intensive intestinal and hepatic mononuclear immune cell infiltration, lamina propria expansion and intestinal villus detachment and shortening (p < .05). In conclusion, despite containing lower glucosinolate, phytic acid, phenolic compounds and tannins well within the recommended thresholds for aquatic animals nutrition, diets containing PCM beyond 25% negatively affected the growth performance and intestine and liver tissue histoarchitecture of juvenile Nile tilapia. However, further complementary studies are needed regarding the effects observed on gut histology and expression of antioxidant enzyme genes.

Antifibrotic effects of hypocalcemic vitamin D analogs in murine and human hepatic stellate cells and in the CCl4 mouse model

Liver cirrhosis is a life-threatening consequence of liver fibrosis. The aim of this study was to investigate the antifibrotic potential of clinically available vitamin D analogs compared to that of calcitriol in vitro and in vivo. Murine hepatic stellate cells, Kupffer cells, and human LX-2 cells were treated with vitamin D analogs, and the profibrotic behavior of these cells was studied. In vivo liver fibrosis was induced using CCl4 until measurable fibrosis was established. Animals were then treated with calcitriol and paricalcitol. Vitamin D and its analogs showed antifibrotic effects in vitro. Treatment with active vitamin D (calcitriol, CAL) and its analogs reduced the protein expression of α-smooth muscle actin (α-SMA) in mHSC. In human LX-2 cells alfacalcidol reduced transforming growth factor-β (TGF-β) induced platelet-derived growth factor receptor-β protein expression and contractility while paricalcitol (PCT), in its equipotent dose to CAL, reduced TGF-β induced α-SMA protein expression, and ACTA2 and TGF-β mRNA expression. No effects of a treatment with vitamin D and its analogs were observed in Kupffer cells. In vivo, PCT-treated mice had significantly lower calcium levels than CAL-treated mice. CAL and PCT reduced the hepatic infiltration of CD11b-positive cells and alanine transaminase levels, while PCT but not CAL significantly inhibited fibrosis progression, with a favorable side effect profile in the CCl4 model. We conclude that hypocalcemic vitamin D analogs should be considered in future studies investigating vitamin D for the treatment of liver fibrosis.

Hepatic infiltration helps predict outcome in dogs with multicentric large B cell lymphoma

Summary Background The five-level WHO staging system is currently the standard for canine lymphomas. In humans, staging for Non-Hodgkin lymphomas has recently evolved to the Lugano classification. In this four-level classification, the splenic infiltration does not induce stage migration from stage III, while hepatic involvement is considered as a more advanced stage together with bone marrow or non-lymphoid organs infiltration. Objective The objective of this pilot study is to evaluate the accuracy of the Lugano staging system, compared to the WHO, in predicting first remission duration (FRD) in dogs. Material and methods Thirty dogs with multicentric large B cell lymphoma, staged according to both systems, and treated with a standard chemotherapy protocol were retrospectively included. Kaplan-Meier curves were established for all stages and compared using a Log Rank test. Results The median FRD was 157 days. Factors significantly associated with shorter FRD in the univariate analysis were substage b (P = 0.024), WHO stage V (P = 0.008) and Lugano stage IV (P = 0.002). Two multivariate Cox models were then compared: the model including substage together with both WHO and Lugano staging was statistically superior to predict FRD as compared to the model including only substage and WHO staging (P = 0.014). Conclusion Taking hepatic involvement into account allows better risk stratification for canine LBCL.

From the Editor’s Desk…: December 2019

From the Editor’s Desk…: December 2019

Evaluation of Dose Related Structural Changes in Sodium-Valproate-Induced Hepatotoxicity and a Possible Protective Role of Vitamin E in Adult Albino Rats

Background: Sodium valproate (SV) is a widely administered antiepileptic drug, although hepatotoxicity is a side effect. Vitamin E (vit. E) being a potent antioxidant agent and essential fat-soluble nutrient that can dramatically decreased this hepatotoxic effect.Aim of the Work: To investigate the histopathological and ultrastructural changes caused by different doses of SV, observe their correlations with liver biomarker levels and assess the defensive role of vit. E against SV-hepatotoxicity.Methods: Sixty male adult albino rats were randomly divided into six groups. Group 1 was treated with normal saline. Group 2 was treated orally with vit. E (100 mg/kg/day). Groups 3, 4 and 5 were treated intraperitoneally with SV at doses of 100, 300 and 500 mg/kg/day, respectively, for 8 consecutive days. Group 6 was treated intraperitoneally with SV (500 mg/kg/day) and orally with vit. E (100 mg/kg/day) for 8 consecutive days. On the ninth day, blood samples were collected to assess the biochemical markers of the liver, and the results were statistically analysed. The rats were deeply anaesthetized and sacrificed. Liver specimens were carefully dissected, and portions were fixed in 10% formalin solution for histopathological examination; others were fixed in 2.5% glutaraldehyde for ultrastructural study.Results: The liver function among the different groups was found to be significantly changed in dose dependent manner. Histopathological examination showed gradual distortion of hepatic lobular architecture and infiltration of lymphocytic cells. Concerning hepatocyte ultrastructure, SV was a destructive compound for most intracellular organelles. This toxicity was most obvious in the groups treated with higher doses; however, concurrent administration of vit. E with SV provided some hepatoprotection.Conclusion: SV is a destructive compound to the liver architecture, especially in high doses. However, SV damage can be attenuated by concurrent administration of vit. E, which considerably decreases most SV-induced hepatotoxicity.

Impact of Rifaximin Therapy on Ischemia/Reperfusion Injury in Liver Transplantation: A Propensity Score–Matched Analysis

Intestinal microbiota is thought to play an important role in hepatic ischemia/reperfusion injury (IRI) after liver transplantation (LT). Rifaximin, a nonabsorbable antibiotic used to treat encephalopathy, exhibits antibacterial activity within the gut. We report the first study examining the impact of pre-LT rifaximin use on reducing hepatic IRI and inflammatory cell infiltration after LT. This retrospective single-center study included adult LT recipients from January 2013 through June 2016. Patients were divided into 2 groups based on duration of rifaximin use before LT: rifaximin group (≥28 days) and control group (none or <28 days). Patients receiving other antibiotics within 28 days of LT and re-LTs were excluded. Outcomes and messenger RNA (mRNA) expression in the graft were compared by 1:1 propensity score-matching and multivariate analyses. On 1:1 matching (n = 39/group), rifaximin patients had lower postoperative serum transaminase levels and lower early allograft dysfunction (EAD; 10.3% versus 33.3%; P = 0.014). Of the matched patients, 8 patients (n = 4/group) had postreperfusion liver biopsies (approximately 2 hours after reperfusion) available for mRNA analysis. Hepatic expression of CD86 (macrophage marker) and cathepsin G (neutrophil marker) was significantly lower in rifaximin patients than controls (P < 0.05). The multivariate analysis included 458 patients. Rifaximin treatment <28 days was identified as an independent risk factor EAD in all patients and those with high Model for End-Stage Liver Disease (MELD) score (MELD ≥35; n = 230). In conclusion, the propensity score-matched and multivariate analyses suggest a therapeutic role of rifaximin in reducing EAD. Pre-LT rifaximin administration exerted a protective function against early liver injury, potentially by suppressing inflammatory cell activation in the graft.

Treatment of acetaminophen-induced liver failure by blocking the death checkpoint protein TRAIL

Acetaminophen (APAP) is one of the most commonly used drugs worldwide, and APAP-induced liver injury is the most frequent cause of acute liver failure in developed countries. However, the mechanisms of APAP-induced hepatotoxicity are not well understood, and treatment options for the disorder are very limited. Here, we show that TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is a major mediator of APAP-induced liver injury in mice, and its blockade markedly ameliorates the liver failure. In APAP-treated mice, TRAIL was expressed in the liver, spleen, and peripheral blood primarily by CD11b+Gr1+ neutrophils. The concentration of soluble TRAIL in the blood, and the frequencies of TRAIL+ leukocytes in the spleen and liver positively correlated with the severity of liver injury. APAP sensitized hepatocytes to TRAIL-induced apoptosis by upregulating the expression of the TRAIL receptor DR5 (death receptor 5), presumably through its transcription factor CHOP (C/EBP homologous protein). Importantly, blocking TRAIL with a soluble DR5-Fc fusion protein (sDR5-Fc) significantly attenuated APAP-induced liver injury, the hepatic infiltration of leukocytes, the levels of inflammatory cytokines, and the mortality of mice. When administered alongside N-acetylcysteine, sDR5-Fc further protected against APAP-induced acute liver injury. Thus, the TRAIL-DR5 signaling pathway plays a key role in APAP-induced liver inflammation and failure, and its blockade represents an effective new strategy to treat the liver disease.

Forkhead Box M1 Transcription Factor Drives Liver Inflammation Linking to Hepatocarcinogenesis in Mice.

Background & AimsLiver inflammation has been recognized as a hallmark of hepatocarcinogenesis. Although Forkhead Box M1 (FoxM1) is a well-defined oncogenic transcription factor that is overexpressed in hepatocellular carcinoma (HCC), its role in liver inflammation has never been explored.MethodsWe generated hepatocyte-specific FoxM1 conditional transgenic (TG) mice by using the Cre-loxP and Tetracycline (Tet)-on systems to induce FoxM1 expression in a hepatocyte-specific and time-dependent manner.ResultsAfter treatment of Tet-derivatives doxycycline (DOX) to induce FoxM1, TG mice exhibited spontaneous development of hepatocyte death with elevated serum alanine aminotransferase levels and hepatic infiltration of macrophages. The removal of DOX in TG mice completely removed this effect, suggesting that spontaneous inflammation in TG mice occurs in a hepatocyte FoxM1-dependent manner. In addition, liver inflammation in TG mice was associated with increased levels of hepatic and serum chemokine (C-C motif) ligand 2 (CCL2). In vitro transcriptional analysis confirmed that CCL2 is a direct target of FoxM1 in murine hepatocytes. After receiving FoxM1 induction since birth, all TG mice exhibited spontaneous HCC with liver fibrosis at 12 months of age. Hepatic expression of FoxM1 was significantly increased in liver injury models. Finally, pharmacologic inhibition of FoxM1 reduced liver inflammation in models of liver injury.ConclusionsHepatocyte FoxM1 acts as a crucial regulator to orchestrate liver inflammation linking to hepatocarcinogenesis. Thus, hepatocyte FoxM1 may be a potential target not only for the treatment of liver injury but also for the prevention toward HCC.

Liquiritigenin and liquiritin alleviated monocrotaline-induced hepatic sinusoidal obstruction syndrome via inhibiting HSP60-induced inflammatory injury

Hepatic sinusoidal obstruction syndrome (HSOS) is a life-threatening liver disease caused by the damage to liver sinusoidal endothelial cells (LSECs). Liquiritigenin and liquiritin are two main compounds in Glycyrrhizae Radix et Rhizoma (Gan-cao). Our previous study has shown that both liquiritigenin and liquiritin alleviated monocrotaline (MCT)-induced HSOS in rats via inducing the activation of nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant signaling pathway. This study aims to further investigate whether inhibiting liver inflammatory injury also contributed to the liquiritigenin and liquiritin-provided alleviation on MCT-induced HSOS. The results of serum alanine/aspartate aminotransferases (ALT/AST) activities and total bilirubin (TBil) amount, liver histological evaluation, scanning electron microscope observation and hepatic metalloproteinase-9 (MMP9) expression showed that liquiritigenin and liquiritin both alleviated MCT-induced HSOS in rats. Liquiritigenin and liquiritin reduced the increased liver myeloperoxidase (MPO) activity, mRNA expression of pro-inflammatory factors, hepatic infiltration of immune cells, hepatic toll-like receptor 4 (TLR4) expression and nuclear factor κB (NFκB) nuclear accumulation induced by MCT in rats. Furthermore, liquiritigenin and liquiritin attenuated MCT-induced liver mitochondrial injury, increased the decreased Lon protein expression and reduced the release of heat shock protein 60 (HSP60). Moreover, liquiritigenin and liquiritin also reduced NFκB nuclear accumulation and decreased the elevated cellular mRNA expression of NFκB-downstream pro-inflammatory cytokines induced by HSP60 in macrophage RAW264.7 cells. In conclusion, our study revealed that both liquiritigenin and liquiritin alleviated MCT-induced HSOS by inhibiting hepatic inflammatory responses triggered by HSP60.

Therapeutic effect of P-coumaric acid in the experimentally infected rats with Salmonella typhi

Salmonella enterica subspecies enteric serotype typhi which infect the human and cause typhoid fevers (systemic infection). Experimental infection with Salmonella typhi are very important because they explains the pathogenicity of Salmonella typhi through a histopathological findings. The herbal inhibitors like P-Coumaric acid explain anti-inflammatory effect and have a selective effect against type III secretion system formation of Salmonella typhi. our study includes 15 adult male albino rats were divided into three groups each group composed from 5 rats. A group: were orally challenged with 1ml of 3X106 CFU of Salmonella typhi for one day. Group B: were also orally administered with 1ml of 3X106 CFU of Salmonella typhi, additionally, they also orally received 5ml of 100mg/kg of P-Coumaric acid for three consecutive days. Group C: serving as control group, they orally administered with 5ml of normal saline. All the animals are sacrificed after five days post infection, liver, small intestine and colon specimens were taken for histopathological investigation. the histological sections of liver and small intestine of group A animals showed pathological changes like degeneration of hepatic cells and inflammatory cell infiltration of the lamina properia in small intestine, while the group B animals showed normal liver and small intestine sections like the control group

Hepatoprotective Effect of Tagetes Erecta L. Extract on Carbon Tetrachloride Induced Hepatotoxicity in Rats

Tagetes erecta L. is a widespread garden plant that is commonly known as the marigold, and it is widely used as a medicinal herb for its anti-inflammatory, analgesic, and anti-edematous properties, which are important for phytotherapeutic, dermatological and cosmetic applications. In this study, the protective effects of water extract of Tagetes erecta L. against liver damage were evaluated in carbon tetrachloride (CCl4)-induced chronic hepatotoxicity in rats. The results showed that the treatment of extract significantly lowered the CCl4-induced serum levels of hepatic enzyme markers (GOT, GPT, ALP, and total bilirubin Liver histopathology showed that extract reduced the incidence of liver lesions including hepatic cells cloudy swelling, lymphocytes infiltration, hepatic necrosis, and fibrous connective tissue proliferation induced by CCl4 in rats. Therefore, the results of this study suggest that Tagetes erecta L. Extract could protect liver against the CCl4-induced oxidative damage in rats, and this hepatoprotective effect might be contributed to its modulation on detoxification enzymes and its antioxidant and free radical scavenger effects.&#x0D; Keywords: Tagetes erecta L., Carbon Tetrachloride, Liver Damage, Hepato-protection

Redox regulation of hepatic NLRP3 inflammasome activation and immune dysregulation in trichloroethene-mediated autoimmunity

Trichloroethene (TCE) exposure is associated with the development of various autoimmune diseases (ADs), including autoimmune hepatitis (AIH) and systemic lupus erythematosus (SLE), potentially through the generation of excessive reactive oxygen and nitrogen species (RONS; oxidative stress). However, the mechanisms by which oxidative stress contributes to these TCE-mediated ADs are not fully understood, and are the focus of current investigation. Female MRL+/+ mice were treated with TCE along with or without antioxidant N-acetylcysteine (NAC) for 6 weeks (TCE, 10 mmol/kg, i. p., every 4th day; NAC, 250 mg/kg/day via drinking water). TCE-treated mice had elevated antinuclear antibodies (ANA) and 4-hydroxynonenal (HNE)-specific circulating immune complexes, suggesting the association of TCE-induced oxidative stress with autoimmune response. In addition, TCE exposure led to prominent lobular inflammation with sinusoid dilation, increased sinusoidal cellularity and increased staining for proliferating cell nuclear antigen (PCNA), confirming inflammatory and hepatocellular cell proliferation. Importantly, TCE exposure resulted in the activation of hepatic inflammasome (NLRP3 and caspase-1) and up-regulation of pro-inflammatory cytokine IL-1β, and these changes were attenuated by NAC supplementation. TCE treatment also led to dysregulation of hepatic immune response as evident from markedly increased hepatic lymphocyte infiltration (especially B cells) and imbalance between Tregs (decreased) and Th17 cells (increased). Interestingly, TCE-mediated dysregulation of various hepatic and splenic immune cells was also effectively attenuated by NAC. Taken together, our findings provide evidence for TCE-mediated inflammasome activation, infiltration of various immune cells, and skewed balance of Treg and Th17 cells in the liver. The attenuation of TCE-mediated hepatic inflammasome activation and immune responses by NAC further supports a critical role of oxidative stress in TCE-mediated inflammation and autoimmunity. These novel findings could help in designing therapeutic strategies for such ADs.

Retinoic acid modulates IL-4, IL-10 and MCP-1 pathways in immune mediated hepatitis and interrupts CD4+ T cells infiltration

AimsImmune mediated liver injury includes activation of different immune pathways that requires various modalities to control their consequences. The current study involves evaluation of retinoic acid (RA) modulatory effects on immune responses induced in concanavalin A (ConA) model of acute hepatitis. Main methodsMice were divided as follows: Control group; RA group: received 35 mg/kg RA; ConA group: received 15 mg/kg ConA; ConA + RA group: received ConA and RA as described. Liver function biomarkers were measured in addition to malondialdehyde as lipid peroxidation biomarker. Liver tissue sections were scored for necro-inflammation, neutrophils infiltration, CD4+ T cells infiltration and NF-κb positive cells. Effect on hepatic levels of TNF-α, IL-4, IL-10 and MCP-1 was evaluated as well. Key findingsInjection of RA before ConA significantly (p < 0.001) decreased ALT, AST and LDH levels compared to their levels in ConA group. Hepatic infiltration of neutrophils and CD4+ T cells was markedly (p < 0.001) reduced by RA. Hepatic injury, necrosis and expression of NF-κb were significantly decreased by RA when injected before ConA challenge. A significant decrease in the measured cytokines TNF-α and IL-4 was observed in ConA + RA group in addition to a decrease in MCP-1 level. On the other hand, IL-10 was significantly increased in the latter group compared to ConA group. SignificanceRA can protect against ConA-induced hepatitis through: interrupting early inflammatory response as neutrophils, monocytes and CD4+ T cells infiltration, modulating IL-4 level and subsequent production of TNF-α and NF-κb activation, mitigating second inflammatory responses through increasing IL-10 liver production.

Integrin β1-enriched extracellular vesicles mediate monocyte adhesion and promote liver inflammation in murine NASH

Hepatic recruitment of monocyte-derived macrophages (MoMFs) contributes to the inflammatory response in non-alcoholic steatohepatitis (NASH). However, how hepatocyte lipotoxicity promotes MoMF inflammation is unclear. Here we demonstrate that lipotoxic hepatocyte-derived extracellular vesicles (LPC-EVs) are enriched with active integrin β1 (ITGβ1), which promotes monocyte adhesion and liver inflammation in murine NASH. Hepatocytes were treated with either vehicle or the toxic lipid mediator lysophosphatidylcholine (LPC); EVs were isolated from the conditioned media and subjected to proteomic analysis. C57BL/6J mice were fed a diet rich in fat, fructose, and cholesterol (FFC) to induce NASH. Mice were treated with anti-ITGβ1 neutralizing antibody (ITGβ1Ab) or control IgG isotype. Ingenuity® Pathway Analysis of the LPC-EV proteome indicated that ITG signaling is an overrepresented canonical pathway. Immunogold electron microscopy and nanoscale flow cytometry confirmed that LPC-EVs were enriched with activated ITGβ1. Furthermore, we showed that LPC treatment in hepatocytes activates ITGβ1 and mediates its endocytic trafficking and sorting into EVs. LPC-EVs enhanced monocyte adhesion to liver sinusoidal cells, as observed by shear stress adhesion assay. This adhesion was attenuated in the presence of ITGβ1Ab. FFC-fed, ITGβ1Ab-treated mice displayed reduced inflammation, defined by decreased hepatic infiltration and activation of proinflammatory MoMFs, as assessed by immunohistochemistry, mRNA expression, and flow cytometry. Likewise, mass cytometry by time-of-flight on intrahepatic leukocytes showed that ITGβ1Ab reduced levels of infiltrating proinflammatory monocytes. Furthermore, ITGβ1Ab treatment significantly ameliorated liver injury and fibrosis. Lipotoxic EVs mediate monocyte adhesion to LSECs mainly through an ITGβ1-dependent mechanism. ITGβ1Ab ameliorates diet-induced NASH in mice by reducing MoMF-driven inflammation, suggesting that blocking ITGβ1 is a potential anti-inflammatory therapeutic strategy in human NASH. Herein, we report that a cell adhesion molecule termed integrin β1 (ITGβ1) plays a key role in the progression of non-alcoholic steatohepatitis (NASH). ITGβ1 is released from hepatocytes under lipotoxic stress as a cargo of extracellular vesicles, and mediates monocyte adhesion to liver sinusoidal endothelial cells, which is an essential step in hepatic inflammation. In a mouse model of NASH, blocking ITGβ1 reduces liver inflammation, injury and fibrosis. Hence, ITGβ1 inhibition may serve as a new therapeutic strategy for NASH.

Laparoscopic Sleeve Gastrectomy on the Horizon as a Promising Treatment Modality for NAFLD.

Nonalcoholic fatty liver disease (NAFLD) is viewed as the hepatic manifestation of the metabolic syndrome with hepatic fatty infiltration is the common liver pathology. NAFLD can lead to nonalcoholic steatohepatitis (NASH), liver cirrhosis, liver cell failure, and liver malignancy.The present work aims to prospectively study the histological changes that occur in NAFLD obese patients 1-year post-laparoscopic sleeve gastrectomy (LSG) based on standardized NAS (NAFLD activity score). This prospective study included 94 obese patients who underwent laparoscopic sleeve gastrectomy. Intraoperative wedge liver biopsy was taken from all patients with a follow-up liver biopsy at 12months after the operation. LSG produced a marked reduction in body weight with a mean reduction in BMI from 44.54 + 5.45 to 34.23 + 2.66kg/m2 at 12months. There were statistically highly significant improvements regarding metabolic comorbidities, blood pressure, lipid profile, and HbA1C at 12-month post-LSG (P < 0.001). The current study showed a highly statistically significant improvement at 1-year post LSG regarding steatosis grade, hepatocyte ballooning, lobular inflammation as well as fibrosis stage (P < 0.001). Moreover, the present study showed that NAS score significantly decreased from 5.20 + 1.96 at baseline to 2.63 + 1.55 at 1-year follow-up (P < 0.001). Our relatively large patient cohort shows a significant improvement of steatosis, steatohepatitis, and fibrosis at a 1-year follow-up. LSG can lead to resolution of NAFLD, and it may be in the near future another routine indication for bariatric surgery.

A novel AMPK activator improves hepatic lipid metabolism and leukocyte trafficking in experimental hepatic steatosis

A novel AMP-activated protein kinase (AMPK) activator, IMM-H007 (H007), has been reported to reduce serum lipid levels and inhibit lipid accumulation in the liver in hyperlipidemic animal models. However, how H007 ameliorates hepatic steatosis and inflammation remains unknown. In the present study, H007, at 200 mg/kg, reduced hepatic lipid levels and the levels of collagenous fiber in the liver in high-fat diet (HFD)-fed hamsters compared to those in the HFD group. Meanwhile, compared to the controls, H007 significantly inhibited sterol-regulatory element binding protein (SREBP)-1c and acetyl CoA carboxylase (ACC) expression by upregulating the AMPK activity, suppressing the saturated fatty acid accumulation and increasing polyunsaturated fatty acid synthesis in the liver. Compared to the controls, H007 treatment inhibited the expression of monocyte chemotactic protein (MCP-1) in fatty acid-treated HepG2 cells; suppressed leukocyte adherence and rolling on the liver microvasculature; and suppressed hepatic macrophage infiltration. H007 also suppressed the expression of nuclear factor-κB (NF-κB) p65 in fatty acid- and lipopolysaccharide-treated HepG2 cells compared to that in the controls by activating AMPK. These data suggested that H007 had a beneficial effect by improving the lipid composition in the liver and inhibiting inflammatory cell trafficking in the development of nonalcoholic fatty liver disease.

Association Between Visfatin and Hepatic Steatosis in the General Population During Long-Term Follow-Up.

The aim of this study was to investigate any association between the adipose tissue-derived protein, visfatin, and non-alcoholic fatty liver disease (NAFLD) and its potential long-term impact on hepatic steatosis. A cross-sectional study including 2429 randomly selected subjects was performed in 2002. Later, 403 subjects were re-evaluated in 2013. Serum visfatin concentrations were determined by sandwich enzyme-linked immunosorbent assay. Phenotyping included abdominal ultrasonography, anthropometric data, and laboratory investigations. No association was found between circulating visfatin levels and the presence of NAFLD at baseline (2002: p=0.0967) or during follow-up (2013: p=0.1312). However, a significant increase in visfatin levels in relation to the level of steatosis was seen during follow-up (p<0.0001). During the more than 10-year follow-up, the metabolic status of the study subjects worsened, with a significant increase in body mass index (BMI) (p<0.0001), waist-to-hip ratio (p<0.0001), triglycerides (TG) (p<0.0001), low-density lipoprotein (p=0.0305), homeostasis model assessment (p<0.0001), and presence of diabetes (p<0.0001). This change was accompanied by an increase in serum visfatin levels, which showed a weak correlation with BMI (p<0.0001, r=0.27586) and presence of diabetes (p<0.0043, r=0.14188). A statistically significant correlation between leucocyte numbers and serum visfatin concentration (p<0.0001, r=0.25615) was found. We found no association between visfatin levels and the presence or absence of NAFLD or the degree of hepatic fatty infiltration at baseline. There was a strong correlation between serum visfatin concentrations and the number of leucocytes, which may suggest a proinflammatory role for visfatin.

The involvement of DAMPs-mediated inflammation in cyclophosphamide-induced liver injury and the protection of liquiritigenin and liquiritin

Cyclophosphamide (CPA) is a chemotherapeutic drug widely used in the treatment of breast cancer or leukemia in clinic. However, CPA was reported to have hepatotoxicity. This study aims to observe the engaged mechanism of CPA-induced liver injury in mice and the protection of liquiritin (LQ) and liquiritigenin (LG). Liver sinusoidal endothelial injury induced by CPA (20, 40 mg/kg) in mice was evidenced by the elevated hepatic metalloproteinase-9 (MMP-9) expression, and the results from liver histological evaluation and scanning electron microscope observation. CPA increased hepatic infiltration of neutrophils, liver myeloperoxidase (MPO) activity, serum interleukin-6 (IL-6) content, hepatic IL-6 mRNA expression, toll-like receptor-4 (TLR4) expression and nuclear factor κB (NFκB) activation in mice. Elevated serum contents of damage associated molecular patterns (DAMPs) including high mobility group box 1 (HMGB1), heat shock protein 60 (HSP60) and glucose-regulated protein 94 (Grp94) were found in mice treated with CPA. Liver sinusoidal endothelial injury and inflammation induced by CPA were diminished in TLR4 knock-out mice. LG and LQ (40, 80 mg/kg) both ameliorated liver sinusoidal endothelial injury, and reduced the increased hepatic infiltration of neutrophils, MPO activity, hepatic IL-6 mRNA expression and NFκB activation induced by CPA. In summary, these results indicate that TLR4-NFκB-mediated inflammatory injury initiated by DAMPs was critically involved in CPA-induced hepatotoxicity. LG and LQ alleviated CPA-induced liver sinusoidal endothelial injury and inflammatory injury in mice.

Brilliant blue G protects against brain and liver tissue damage during systemic endotoxemia in rats treated with lipopolysaccharide

Brilliant blue G or Coomassie blue is a purinergic P2X7 receptor antagonist. In this study, the effect of brilliant blue G on serum oxidative stress and brain and liver tissue damage was studied in rats with lipopolysaccharide (LPS)-induced systemic inflammation. Rats were treated with intraperitoneal (i.p.) injection of Escherichia coli LPS (300 μg/kg) alone or together with brilliant blue G at 5 or 10 mg/kg and euthanized 4 h thereafter. Serum malondialdehyde, nitric oxide, paraoxonase 1 (PON-1) activity, cholinesterase activity, and glucose were determined. In addition, brain and liver histopathology, and caspase-3 and glial fibrillary acidic protein (GFAP) immunostaining were done. Results indicated that LPS produced a significant elevation in serum malondialdehyde and nitric oxide concentrations along with markedly decreased PON-1 activity and glucose. LPS caused neuronal degeneration in the cerebral cortex and hippocampus, increased caspase-3, and decreased GFAP immunostaining in the cerebral cortex. Vacuolar degeneration and inflammation were observed in the liver of LPS-treated rats. The administration of brilliant blue G decreased serum malondialdehyde by 34.5–35.2% and nitric oxide concentrations by 27.4–35.6%, respectively, while increasing PON-1 activity by 46.3–86.7% and serum glucose level by 24.8%. Moreover, brilliant blue G inhibited serum cholinesterase activity by 38.1–42% compared with the LPS control group. Brilliant blue G attenuated the neuronal degeneration, the increase in caspase-3 activity, and the decrease in GFAP-positive astrocytes produced by LPS. It also decreased hepatic cellular infiltration and congestion. These results indicate that brilliant blue G is able to ameliorate the brain and liver tissue damage during LPS-induced systemic inflammation and suggest a potential therapeutic use of brilliant blue G to prevent organ damage during systemic endotoxemia.

Amygdalin (Vitamin B17) pretreatment attenuates experimentally induced acute autoimmune hepatitis through reduction of CD4+ cell infiltration

BackgroundAutoimmune hepatitis (AIH) is an immune-mediated inflammation of the liver characterized by disorganized hepatic parenchyma and inflammatory cell infiltration. Although the increased incidence of AIH, the development of novel therapeutic strategies are impeded by the poor understanding of the accompanied detailed immunopathogenic changes. CD4+ T cells are key mediators of inflammatory cell infiltration in initial phases of liver injuries like AIH. The distribution of CD4+ cells and the histopathological changes accompanying Con A-induced AIH were investigated together with the postulated protective effect of Amygdalin (Amg.). Materials and methods30 adult male mice were divided into three groups; control, AIH and AIH-Amg. groups. AIH was induced by a single intravenous injection of Concanavalin A (Con A) (15 mg/kg). The AIH-Amg. group received Amg. 5 mg/kg intraperitoneally once a week for three weeks. Blood samples were examined for ALT and AST. MDA, SOD, and GSH were determined in hepatic homogenates. Liver section stained with hematoxylin and eosin, Masson trichrome and CD4+ immune stain were examined by light and electron microscopy. ResultsAIH group showed a significant increase in levels of ALT, AST and MDA and a significant decline in SOD and GSH compared to the controls. The liver tissue showed distorted hepatic architecture with intercellular hemorrhage, necrosis, and inflammatory cell infiltration. The area percent of CD4+ immune staining was significantly increased. Electron microscopic examination showed massive cellular degenerative changes. Amg. pretreatment in AIH-Amg. group significantly reversed these changes. ConclusionAIH induced CD4+ cells infiltration in the liver with subsequent liver tissue damage. Amg. pretreatment inhibited CD4+ cell infiltration and protected the liver tissue. This finding suggests that Amg. could be a therapeutic agent in the management of AIH.