Lower Serum Alanine Aminotransferase Research Articles

The relationship between urinary genistein levels and serum alanine aminotransferase levels in adults in the USA: National Health and Nutrition Examination Survey 1999–2010

Genistein, a phytoestrogen with similarities to female sex hormones, has been shown to protect against oxidative stress and fibrosis in nonalcoholic fatty liver injury in animal studies. However, few studies have examined genistein's effects on liver function in humans. We analyzed data from the National Health and Nutrition Examination Survey from 1999 to 2010. Individuals younger than 21 years, with viral hepatitis, or with serum alanine aminotransferase (ALT) at the extremes of distribution (5% on each extreme) were excluded. Urinary genistein was normalized by urinary creatinine levels. The relationship between normalized urinary genistein (nUG) and serum ALT was examined using linear regression models with and without adjustment for potential confounders, and the differential effect of sex was examined using an interaction term. Of the 9864 participants, 52% were female, 50% were White, 24% were elderly, 36% had hypertension, 12% had diabetes, and 8.1% were heavy alcohol drinkers. Serum ALT was significantly lower in the highest quartile compared with the lowest quartile of nUG (22.3 vs. 23.5 U/l; P<0.001). In adjusted models, individuals in the highest quartile had 0.75 U/l lower ALT levels than those in the lowest quartile (P=0.02). We found a significant difference in ALT levels between the lowest and highest quartiles of nUG in males, but not in females (difference in differences=1.77 U/l, interaction P=0.04). We found a statistically significant association between higher nUG and lower serum ALT in males, but not in females. The sex-specific role of genistein in mitigating liver disease merits further study.

CXCR2 Regulates Hepatic Injury after Bile Duct Ligation by Altering Neutrophil Migration

IntroductionCXC chemokines and their receptor, CXCR2, are important components for progression of hepatic inflammation. However, the contribution of CXCR2 for cholestatic liver injury have not been elucidated. Neutrophil recruitment in the liver and their releasing reactive oxygen species during cholestasis contribute to liver injury. In the current study, we sought to determine the role of CXCR2 in hepatic injury after bile duct ligation (BDL).MethodsWild‐type (WT) and CXCR2−/− mice were subjected to BDL to induce cholestatic liver injury. Serum and liver samples were obtained 14 days after BDL. Serum levels of alanine amino transferase (ALT) and liver myeloperoxidase content (MPO) were measured. Liver sections were stained with hematoxylin and eosin for assessing the extent of liver necrosis. Neutrophils in liver tissue was determined by immunonhistochemical staining for Ly‐6G.ResultsAfter BDL, CXCR2−/− mice had significantly lower serum ALT level and less necrotic area than WT mice. There was no difference in MPO level in the whole liver, however, the distribution of recruited neutrophils was different between WT and CXCR2−/− mice. In WT mice, neutrophils selectively accumulated to necrotic areas in liver parenchyma and infiltrated into the peri‐biliary lesions. In contrast, in CXCR2−/− mice, neutrophils were randomly scattered in the liver parenchyma, with less concentration of neutrophils in necrotic areas and peri‐biliary lesions.ConclusionThe data suggest that CXCR2 regulates liver injury after BDL by directing neutrophil migration to specific sites of hepatocyte and biliary cell injury.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

PDE4 Inhibitor Rolipram Mitigates Liver Ischemia Reperfusion Injury in a Rat Model by Preventing Neutrophil Accumulation

BACKGROUND/INTRODUCTIONIschemia‐reperfusion (IR) injury is caused by a transient loss of blood flow (ischemia) followed by return of blood flow (reperfusion), which initiates an inflammatory response. IR injury occurs in organ transplantation, sepsis, respiratory failure, hemorrhage, trauma, stroke, cardiac arrest, and shock. Ischemia reperfusion, as a primary cause of post graft non‐function, is a limiting factor in the success of liver transplantation. Due to a crucial shortage of donor livers, there are about 17,000 people on the waiting list for a liver transplant, and 1,500 a year will die while waiting for an organ (American Liver Foundation). Pharmacological attenuation of the IR‐induced inflammatory response could improve graft function and survival. We investigated the potential of using the phosphodiesterase (PDE) inhibitor, rolipram, in hepatic ischemia reperfusion injury. PDEs are the only enzymes that can hydrolyze cyclic adenosine monophosphate (cAMP). Higher cAMP levels can initiate and sustain anti‐inflammatory signaling. Therefore, we hypothesized that suppression of the inflammatory response by rolipram would reduce liver damage following IR injury.METHODSMale Sprague‐Daley rats were randomized to six groups (n=6–8 per group): 1) sham, 2) sham + 3 mg/kg rolipram, 3) 40% ischemia reperfusion, 4) 40% ischemia reperfusion + 3 mg/kg rolipram, 5) 70% ischemia reperfusion, and 6) 70% ischemia reperfusion + 3 mg/kg rolipram. Two different levels of severity of IR injury were achieved by clamping either 40% (left lobe) or 70% (left and median lobes) of the liver. In both cases, occlusion was for 1 hour followed by 3 hours of reperfusion. Treatment was given I.V. at the end of the ischemic period. Blood and tissue were collected. Blood chemistry was assayed on a Vetscan 2 (Abaxis). Paraffin embedded liver sections were stained for H&amp;E (morphology), intercellular adhesion molecule 1 (ICAM‐1) (neutrophil adhesion molecule), and chloroacetate esterase (CAE, stains neutrophils). Liver myeloperoxidase (MPO, from neutrophils) and tumor necrosis factor alpha (TNF‐α) and serum hyaluronate (a marker of vascular damage) were assessed by ELISA.RESULTSBlood chemistry revealed that IR significantly (p=0.01) elevated serum alanine aminotransferase (ALT), an indicator of liver damage, in both the 40% and 70% models. Rolipram lowered serum ALT levels in the 40% model. Inflammation‐induced neutrophil infiltration is a major source of IR damage. Histology showed areas of coagulative necrosis and increased neutrophil infiltration, which were ameliorated by rolipram, in both 40% and 70% models. Liver ICAM‐1 was reduced by rolipram treatment in both models. MPO was significantly increased by 70% but not 40% IR, and rolipram did not lower it. Liver TNF‐α showed no difference between sham and IR groups, but rolipram seemed to lower it independently of IR. Rolipram treatment showed a trend towards decreasing serum hyaluronan in both models.CONCLUSIONMechanisms still remain to be determined, but overall, these data suggest that inhibition of PDE reduces hepatic IR injury by decreasing neutrophil recruitment. We conclude that rolipram might have therapeutic potential in ischemia reperfusion injury.Support or Funding InformationSupported by grants from NIH and Veterans AdministrationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Serum alanine aminotransferase level and liver-related mortality in patients with chronic hepatitis B: A large national cohort study.

The serum alanine aminotransferase (ALT) level has been used to identify at-risk patients with chronic hepatitis B (CHB) who need antiviral therapy. However, the level associated with increased liver-related mortality requiring active treatment is still unclear. We used a Health Examination Cohort of the National Health Insurance Service of Korea that included approximately 0.5 million individuals aged 40-79years. In total, 12486 patients with CHB and no other concurrent liver disease were enrolled, and patients' liver-related mortality, including that owing to liver cancer, was investigated over 9years. The serum ALT level was correlated positively with liver-related mortality. The rates in men were 0.14, 0.17, 0.24, 0.57, 0.63 and 0.85 per 100 person-years (%) for serum ALT levels of <20, 20-29, 30-39, 40-49, 50-79 and ≥80U/L, respectively, and the corresponding liver-related mortality rates in women were 0.03%, 0.09%, 0.12%, 0.63%, 0.65% and 0.32%. In patients with ALT levels of 40-79U/L, the liver-related mortality rates were 0.60% in men and 0.64% in women, which were similar to the overall mortality rate of age- and sex-matched subjects without CHB (0.69%). The best cut-off values for liver-related mortality prediction were >34U/L in men and >30U/L in women. The liver-related mortality rate increased significantly, even in CHB patients with relatively low serum ALT levels. Careful monitoring or earlier antiviral therapy should be considered for patients aged >40years with serum ALT levels above the upper limit of normal.

Effects of Curcuminoids Plus Piperine on Glycemic, Hepatic and Inflammatory Biomarkers in Patients with Type 2 Diabetes Mellitus: A Randomized Double-Blind Placebo-Controlled Trial

Curcuminoids have been shown to reduce glycemia and related complications in diabetes. In the present study, we evaluated the impact of curcuminoids plus piperine administration on glycemic, hepatic and inflammatory biomarkers in type 2 diabetes (T2D) patients. T2D patients aged 18-65 years were enrolled in a randomized double-blind placebo-controlled trial and randomly allocated to standard-of-care treatment and dietary advises plus either curcuminoids (daily dose of 500 mg/day co-administered with piperine 5 mg/day) or placebo for a period of 3 months. Glycemic, hepatic and inflammatory parameters were measured at baseline and final conditions. A total of 100 subjects (50 in each group) completed the 3-month period of trial. A significant reduction was found in serum levels of glucose (-9±16 mg/dL vs. -3±11 mg/dL in curcuminoids and placebo groups, respectively; p=0.048), C-peptide (-0.6±0.8 ng/mL vs. 0.02±0.6 ng/mL; p<0.001) and HbA1c (-0.9±1.1% vs. -0.2±0.5%; p<0.001) after curcuminoids supplementation versus placebo group. Additionally, participants in the intervention group showed lower serum alanine aminotransferase (-2±6 vs. -1±5; p=0.032) and aspartate aminotransferase (-3±5 vs. -0.3±4; p=0.002) levels compared with the placebo group. Finally, no significant differences in high-sensitivity C-reactive protein (hs-CRP) concentrations were observed between curcuminoids and placebo groups (p>0.05). The results of the present trial revealed a beneficial effect of curcuminoids plus piperine supplementation on glycemic and hepatic parameters but not on hs-CRP levels in T2D patients.

MiR-494 up-regulates the PI3K/Akt pathway via targetting PTEN and attenuates hepatic ischemia/reperfusion injury in a rat model.

A rat HIRI model was constructed and treated with an intraperitoneal injection of agomir-miR-494 or agomir-NC (negative control) for 7 days after the surgery. The pathophysiological changes in sham-operated rats, HIRI, HIRI + agomir-miR-494, and HIRI + agomir-NC were compared. The effect of miR-494 was also assessed in an H2O2-induced apoptosis model. Hepatic AML12 cells were transfected with mimics NC or miR-494 mimics, followed by 6-h H2O2 treatment. Cell proliferation and apoptosis were detected by CCK8 assay and flow cytometry, respectively. Further, the miR-494 target gene was identified by luciferase reporter assay, and verified both in vitro and in vivo experiments. The activity of AKT pathway was further analyzed in vivo by Western blot. HIRI + agomir-miR-494 rats exhibited significantly higher miR-494 expression, lower serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and glutamate dehydrogenase (GLDH) level, lower hepatic MDA, TOA, and OSI, alleviated hepatic necrosis, reduced hepatocyte apoptosis, and decreased expression of apoptosis-related proteins, when compared with HIRI + agomir-NC rats (P<0.05 or 0.01). After H2O2 treatment, AML-12 cells transfected with miR-494 mimics had significantly higher proliferation and lower apoptosis rate compared with mimics NC group (P<0.01). PTEN was identified as an miR-494 target gene. PTEN expression was significantly down-regulated in AML12 cells transfected with miR-494 mimics, and was up-regulated by treatment of miR-494 inhibitor (P<0.01). Moreover, HIRI + agomir-miR-494 rats exhibited significantly lower PTEN expression, and higher p-AKT, p-mTOR, and p-p70S6K levels compared with HIRI + agomir-NC rats. Therefore, miR-494 protected rats against hepatic ischemia/reperfusion (I/R) injury through down-regulating its downstream target gene PTEN, leading to the activation of PI3K/AKT signaling pathway.

Berberine ameliorates non-alcoholic steatohepatitis in ApoE-/- mice.

The aim of the present study was to explore the protective effects of Berberine (BBR) against non-alcoholic steatohepatitis (NASH). Male 4-week-old C57BL/6J Apolipoprotein E-deficient (ApoE−/−) mice were divided into the following three groups, which were given different diets: Normal chow diet (SC group); high-fat high-cholesterol diet (HFHC group); and HFHC diet supplemented with BBR (BBR group). Serum biochemical indicators of hepatic function and histological liver tissue changes were evaluated. The expression of neutrophil elastase (NE) and genes involved in the inflammatory response was measured. ApoE−/− mice fed a HFHC diet for 12 weeks developed NASH, characterized by steatosis and liver inflammation. Body weight, and serum triglyceride and cholesterol levels were markedly reduced by BBR. BBR supplementation significantly lowered serum alanine aminotransferase and aspartate aminotransferase levels in mice with HFHC diet-induced NASH, and significantly downregulated hepatic expression and activity of NE, whereas α1-antitrypsin (α1-AT) expression was significantly recovered by BBR (all P<0.05 vs. the HFHC group). Furthermore, treatment with BBR induced a significant reduction in the expression of key genes, including phospoinositide 3-kinase, nuclear factor-κB and interleukin-8, in the C-X-C chemokine receptor type 4 (CXCR4) signaling pathway (all P<0.05 vs. the HFHC group). These results suggest that BBR alleviates NASH in ApoE−/− mice fed a HFHC diet. Restoration of the balance of NE and α1-AT levels, which in turn facilitate the inhibition of the CXCR4 signaling pathways, may be involved in the hepatoprotective effect of BBR. These results indicate that BBR may be a candidate therapeutic agent for the treatment of NASH.

Connexin hemichannel inhibition reduces acetaminophen-induced liver injury in mice

Historically, connexin hemichannels have been considered as structural precursors of gap junctions. However, accumulating evidence points to independent roles for connexin hemichannels in cellular signaling by connecting the intracellular compartment with the extracellular environment. Unlike gap junctions, connexin hemichannels seem to be mainly activated in pathological processes. The present study was set up to test the potential involvement of hemichannels composed of connexin32 and connexin43 in acute hepatotoxicity induced by acetaminophen. Prior to this, in vitro testing was performed to confirm the specificity and efficacy of TAT-Gap24 and TAT-Gap19 in blocking connexin32 and connexin43 hemichannels, respectively. Subsequently, mice were overdosed with acetaminophen followed by treatment with TAT-Gap24 or TAT-Gap19 or a combination of both after 1.5h. Sampling was performed 3, 6, 24 and 48h following acetaminophen administration. Evaluation of the effects of connexin hemichannel inhibition was based on a series of clinically relevant read-outs, measurement of inflammatory cytokines and oxidative stress. Subsequent treatment of acetaminophen-overdosed mice with TAT-Gap19 only marginally affected liver injury. In contrast, a significant reduction in serum alanine aminotransferase activity was found upon administration of TAT-Gap24 to intoxicated animals. Furthermore, co-treatment of acetaminophen-overdosed mice with both peptides revealed an additive effect as even lower serum alanine aminotransferase activity was observed. Blocking of connexin32 or connexin43 hemichannels individually was found to decrease serum quantities of pro-inflammatory cytokines, while no effects were observed on the occurrence of hepatic oxidative stress. This study shows for the first time a role for connexin hemichannels in acetaminophen-induced acute liver failure.

A Comparison of Two Mouse Frailty Assessment Tools.

The mouse clinical frailty index and the mouse frailty phenotype assessment are two recently developed tools used to assess frailty in mice. The objectives of this study were to investigate whether the same mice are identified as frail with both tools and to examine the association of each of the assessment tools with age and frailty-related outcomes. Frailty was measured using both tools in old (~24 months; n = 36) C57BL/6 male mice. After 2 weeks, blood pressure and heart rate were measured and serum samples were collected for analysis of alanine aminotransferase, creatinine, and albumin levels. The mouse frailty phenotype assessment identified no mice as frail but modification of the assessment tool identified six mice as frail. The mouse clinical frailty index identified 16 mice as frail and the agreement between the two scales was 50.0%. Increasing clinical frailty index scores were correlated with low serum alanine aminotransferase, as well as decreased heart rate, and reduced heart rate variance. We conclude that, consistent with equivalent frailty assessment scales in humans, both tools have value but do not necessarily identify the same mice as frail.

Intestinal Alkaline Phosphatase Attenuates Alcohol-Induced Hepatosteatosis in Mice.

Bacterially derived factors from the gut play a major role in the activation of inflammatory pathways in the liver and in the pathogenesis of alcoholic liver disease. The intestinal brush-border enzyme intestinal alkaline phosphatase (IAP) detoxifies a variety of bacterial pro-inflammatory factors and also functions to preserve gut barrier function. The aim of this study was to investigate whether oral IAP supplementation could protect against alcohol-induced liver disease. Mice underwent acute binge or chronic ethanol exposure to induce alcoholic liver injury and steatosis±IAP supplementation. Liver tissue was assessed for biochemical, inflammatory, and histopathological changes. An ex vivo co-culture system was used to examine the effects of alcohol and IAP treatment in regard to the activation of hepatic stellate cells and their role in the development of alcoholic liver disease. Pretreatment with IAP resulted in significantly lower serum alanine aminotransferase compared to the ethanol alone group in the acute binge model. IAP treatment attenuated the development of alcohol-induced fatty liver, lowered hepatic pro-inflammatory cytokine and serum LPS levels, and prevented alcohol-induced gut barrier dysfunction. Finally, IAP ameliorated the activation of hepatic stellate cells and prevented their lipogenic effect on hepatocytes. IAP treatment protected mice from alcohol-induced hepatotoxicity and steatosis. Oral IAP supplementation could represent a novel therapy to prevent alcoholic-related liver disease in humans.

Analysis of the effect of Qizhuyigan on liver function in a mouse model of immunological liver injury

Purpose : To assess the protective effect of Qizhuyigan capsules containing an herbal mixture on liver function in a mouse model of immunological liver injury and to explore the mechanism of action. Methods : One hundred and twenty mice were randomly divided into four groups: control, test, bifendate, and Qizhuyigan. Immunological liver injury was induced in all groups except the control group. Mice in the control group and the test group were gavaged with 2.5 g/kg tap water, mice in the bifendate group were gavaged with 12 mg/kg bifendate in water, and mice in the Qizhuyigan group were gavaged with 1,000 mg/kg of an aqueous solution containing the contents of a Qizhuyigan capsule. The gavage continued for 10 days. Changes in liver function-related indices, such as the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), malonaldehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor α (TNF-α), and interleukin 6 (IL-6), were assessed. Results : Compared with the test and bifendate groups, the Qizhuyigan group exhibited lower serum ALT (98.3 U/L ± 8.7 U/L vs. 224.7 U/L ± 18.4 U/L vs. 132.8 U/L ± 9.4 U/L); AST (165.5 U/L ± 12.1 U/L vs. 362.6 U/L ± 16.6 U/L vs. 224.6 U/L ± 12.2 U/L); and MDA levels (12.7 ± 2 μmol/L vs. 31.3 ± 6.60 μmol/L vs. 14.4 ± 2.3 μmol/L); a higher SOD level; and reduced TNF-α and IL-6 levels. The differences of the above indices had statistical significance (p < 0.05). Conclusion : Qizhuyigan exerted a protective effect in a mouse model of immunological liver injury. Keywords : Qizhuyigan capsule, Immunological liver injury, Biochemical parameters

Treatment with dimethyl fumarate ameliorates liver ischemia/reperfusion injury.

AIMTo investigate the hypothesis that treatment with dimethyl fumarate (DMF) may ameliorate liver ischemia/reperfusion injury (I/RI).METHODSRats were divided into 3 groups: sham, control (CTL), and DMF. DMF (25 mg/kg, twice/d) was orally administered for 2 d before the procedure. The CTL and DMF rats were subjected to ischemia for 1 h and reperfusion for 2 h. The serum alanine aminotransferase (ALT) and malondialdehyde (MDA) levels, adenosine triphosphate (ATP), NO × metabolites, anti-oxidant enzyme expression level, anti-inflammatory effect, and anti-apoptotic effect were determined.RESULTSHistological tissue damage was significantly reduced in the DMF group (Suzuki scores: sham: 0 ± 0; CTL: 9.3 ± 0.5; DMF: 2.5 ± 1.2; sham vs CTL, P < 0.0001; CTL vs DMF, P < 0.0001). This effect was associated with significantly lower serum ALT (DMF 5026 ± 2305 U/L vs CTL 10592 ± 1152 U/L, P = 0.04) and MDA (DMF 18.2 ± 1.4 μmol/L vs CTL 26.0 ± 1.0 μmol/L, P = 0.0009). DMF effectively improved the ATP content (DMF 20.3 ± 0.4 nmol/mg vs CTL 18.3 ± 0.6 nmol/mg, P = 0.02), myeloperoxidase activity (DMF 7.8 ± 0.4 mU/mL vs CTL 6.0 ± 0.5 mU/mL, P = 0.01) and level of endothelial nitric oxide synthase expression (DMF 0.38 ± 0.05-fold vs 0.17 ± 0.06-fold, P = 0.02). The higher expression levels of anti-oxidant enzymes (catalase and glutamate-cysteine ligase modifier subunit and lower levels of key inflammatory mediators (nuclear factor-kappa B and cyclooxygenase-2 were confirmed in the DMF group.CONCLUSIONDMF improved the liver function and the anti-oxidant and inflammation status following I/RI. Treatment with DMF could be a promising strategy in patients with liver I/RI.

Increasing incidence of elderly-onset autoimmune hepatitis.

Autoimmune hepatitis (AIH) commonly shows bimodal distribution of onset age: at young adulthood and at 50-60 years-of-age. However, in recent times, the incidence of elderly-onset AIH seems to be increasing. This study aimed to investigate whether the incidence of elderly-onset AIH is increasing, and whether these patients show any clinical features different from those observed in younger patients. Data about patients with newly diagnosed AIH visiting the Japanese Red Cross Society Himeji Hospital, Himeji, Hyogo, Japan, were retrospectively collected for the period ranging from January 2010 to May 2016. A total of 71 patients (56 women and 15 men, age 18-88 years) were included in this study. Patients were divided into two cohorts: elderly (≥70 years; n = 28) and adult cohort (15-69 years; n = 43). Demographic and clinical characteristics, biochemical and serological markers, radiological and histological findings, and therapeutic courses were evaluated. The median age of the patients was 65 years, the most frequent range being the 70s (37%), followed by the 60s (25%). The elderly cohort had significantly higher levels of serum immunoglobulin G and antinuclear antibody, lesser hepatitis activity scores, and lesser chance of developing other autoimmune diseases. They tended to have higher C-reactive protein levels and lower serum alanine aminotransferase levels. All patients achieved clinical remission after treatment. This study clearly showed an increase in the incidence of elderly-onset AIH. These patients had some unique characteristics, showing that the development of elderly-onset AIH is influenced by age-associated immune dysfunction called immunosenescence. Geriatr Gerontol Int 2017; 17: 1722-1728.

Microsomal prostaglandin E synthase-1 protects against Fas-induced liver injury.

Microsomal prostaglandin E synthase-1 (mPGES-1) is the terminal enzyme for the synthesis of prostaglandin E2 (PGE2), a proproliferative and antiapoptotic lipid molecule important for tissue regeneration and injury repair. In this study, we developed transgenic (Tg) mice with targeted expression of mPGES-1 in the liver to assess Fas-induced hepatocyte apoptosis and acute liver injury. Compared with wild-type (WT) mice, the mPGES-1 Tg mice showed less liver hemorrhage, lower serum alanine transaminase (ALT) and aspartate transaminase (AST) levels, less hepatic necrosis/apoptosis, and lower level of caspase cascade activation after intraperitoneal injection of the anti-Fas antibody Jo2. Western blotting analysis revealed increased expression and activation of the serine/threonine kinase Akt and associated antiapoptotic molecules in the liver tissues of Jo2-treated mPGES-1 Tg mice. Pretreatment with the mPGES-1 inhibitor (MF63) or the Akt inhibitor (Akt inhibitor V) restored the susceptibility of the mPGES-1 Tg mice to Fas-induced liver injury. Our findings provide novel evidence that mPGES-1 prevents Fas-induced liver injury through activation of Akt and related signaling and suggest that induction of mPGES-1 or treatment with PGE2 may represent important therapeutic strategy for the prevention and treatment of Fas-associated liver injuries.

Contribution and Mobilization of Mesenchymal Stem Cells in a mouse model of carbon tetrachloride-induced liver fibrosis.

Hepatic fibrosis is associated with bone marrow derived mesenchymal stem cells (BM-MSCs). In this study, we aimed to determine what role MSCs play in the process and how they mobilize from bone marrow (BM). We employed a mouse model of carbon tetrachloride(CCl4)-induced liver fibrosis. Frozen section was used to detect MSCs recruited to mice and human fibrotic liver. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) was detected to assess liver function. It was found that MSCs of both exogenous and endogenous origin could aggravate liver fibrosis and attenuate liver damage as indicated by lower serum ALT and AST levels. Stromal cell–derived factor-1 (SDF-1α)/ CXCR4 was the most important chemotactic axis regulating MSCs migration from BM to fibrotic liver. Frozen section results showed that the migration did not start from the beginning of liver injury but occured when the expression balance of SDF-1α between liver and BM was disrupted, where SDF-1α expression in liver was higher than that in BM. Our findings provide further evidence to show the role of BM-MSCs in liver fibrosis and to elucidate the mechanism underlying MSCs mobilization in our early liver fibrosis mice model induced by CCl4.

LXRα gene downregulation by lentiviral-based RNA interference enhances liver function after fatty liver transplantation in rats

Steatotic liver grafts, although accepted, increase the risk of poor posttransplantation liver function. However, the growing demand for adequate donor organs has led to the increased use of so-called marginal grafts. Liver X receptor alpha (LXRalpha) is important in fatty acid metabolism and interrelated with the specific ischemia-reperfusion injury in fatty liver transplantation. This study aimed to investigate whether LXRalpha RNA interference (RNAi) could improve the organ function of liver transplant recipients. Fifty Sprague-Dawley rats were fed with a high-fat diet and 56% alcohol. The livers of these animals had greater than 60% macrovesicular steatosis and were used as liver donors. The experimental donors were treated with 7X107 TU LXRalpha-RNAi-LV of a mixture injection and control donors with negative control-LV vector injection into the portal vein 72 hours before the operation. The effects of LXRalpha-RNAi-LV were assessed by serum aminotransferases, histology, immunostaining, and protein levels. The transcription of LXRalpha mRNA was assessed by reverse transcription-polymerase chain reaction. Compared with controls, LXRalpha RNAi inhibited the expression of LXRalpha at the mRNA (0.53+/-0.03 vs 0.94+/-0.02, P<0.05) and protein levels (0.51+/-0.08 vs 1.09+/-0.12, P<0.05). LXRalpha RNAi also decreased the expressions of sterol regulatory element-binding protein 1c (SREBP-1c) and CD36. LXRalpha RNAi consequently reduced fatty acid accumulation in hepatocytes. Compared with control animals, LXRalpha RNAi-treated group had lower serum alanine aminotransferase, aspartate aminotransferase, interleukin-1beta, and tumor necrosis factor-alpha levels and milder pathologic damages. TUNEL analysis revealed a significant reduction of apoptosis in the livers of rats treated with LXRalpha-RNAi-LV, and overall survival as determined by the Kaplan-Meier method was improved among rats treated with LXRalpha-RNAi-LV (P<0.05). LXRalpha-RNAi-LV treatment significantly downregulated LXRalpha expression and improve steatotic liver graft function and recipient survival after a fatty liver transplantation in rats.

PWE-196 Factors predicting mortality in acute severe gallstone pancreatitis

Introduction The British Society of Gastroenterology (BSG) guidelines state that “Urgent therapeutic ERCP should be performed in patients with acute pancreatitis of suspected or proven gall stone aetiology who satisfy the criteria for predicted or actual severe pancreatitis”. The Cochrane review in 2012 suggested that ERCP does not affect morbidity and mortality and some patients with gallstones in the CBD will pass spontaneously. Aim The aim of this study was to assess predictive factors of mortality and the need for ERCP in severe gallstone pancreatitis. Method Data for patients who presented with acute severe biliary pancreatitis between January 2012 and April 2014 was collected. Data collected included predicted severity (Modified Glasgow score), liver function tests, white cell count, USS, ERCP and MRCP reports. Overall 90 day mortality was also recorded. Results 123 patients had predicted severe pancreatitis. On US 21 patients had a dilated CBD and 16 had CBD stone. 51% of patients (n = 63) had MRCP of which 22 patients had CBD stones. 60 patients had ERCP. In 17% (n = 21) no stone was found. 2 patients (3%) developed post ERCP pancreatitis.11% (n = 14)of the patients admitted with predicted severe acute pancreatitis died. On univariate analysis albumin (p = 0.003), alanine transaminase (ALT) (p= Conclusion Low serum albumin and ALT predicts mortality in severe GSP but ERCP does not and may precipitate further attacks of pancreatitis. Patients who have severe gallstone pancreatitis without deteriorating LFTs or cholangitis should have an MRCP to confirm stones in the CBD prior to ERCP. Disclosure of interest None Declared.

Gravin Scaffolding Protein Mediates Signaling Involved in theRegulation of Hyperlipidemia and Atherosclerosis

Atherosclerosis is a leading health problem and contributes to cardiovascular related mortality. Furthermore, hyperlipidemia is a major atherogenic factor.Understanding the cellular mechanisms leading to atherosclerosis is still lacking. Gravin (AKAP12), an A‐kinase anchoring protein (AKAP), scaffolds and targets various signaling proteins to specific intracellular locations and has been implicated in mediating lipid metabolism. Thus, the purpose of this study was to determine whether the absence of gravin‐mediated signaling would prevent the initiation and progression of atherosclerosis.5 week‐old wild‐type (WT) and gravin‐truncated (gravin‐t/t) mice were subjected to high fat diet (HFD) or normal diet (ND) for 16‐weeks. Cholesterol, triglyceride and VLDL level in serum were significantly decreased in gravin‐t/t HFD compared to WT HFD mice. In addition, systolic and diastolic blood pressure was also lower in gravin‐t/t HFD mice.Furthermore, gravin‐t/t HFD mice showed lower liver‐to‐body weight ratio as well as decreased lipid accumulation and liver damage.These observations are consistent with the lower serum alanine aminotransferase(ALT) and aspartate aminotransferase (AST) levels identified in gravin‐t/t HFD mice. In addition, gravin‐t/t HFD mice had decreased expression of the active form of sterol‐regulatory‐element‐binding protein 2 (SREBP2), which resulted in lower HMG‐CoA reductase (HMGCR) and low‐density lipoprotein receptor (LDLR) expression. Finally, we observed less aortic lipid accumulation in gravin‐t/t HFD mice. Our data indicates that the absence of gravin mediated signaling were resistance to hyperlipidemia and prevented the progression of atherosclerosis.

MicroRNA-155 deficiency attenuates ischemia-reperfusion injury after liver transplantation in mice

Liver ischemia-reperfusion injury (IRI) is a major cause of morbidity and mortality after resection surgery, liver transplantation, and hemorrhagic and septic shock. Mir-155 is upregulated by a broad range of inflammatory mediators, and it has been demonstrated to be involved in both innate and adaptive immune responses. However, the role of mir-155 in liver IRI has never been investigated. In this study, mir-155 deficiency protected mice from liver IRI, as shown by lower serum alanine aminotransferase (ALT) levels and Suzuki scores. Mir-155 deficiency results in the development of M2 macrophages, which respond to IR-induced innate immune stimulation by producing a regulatory inflammatory response with higher level of IL-10, but lower levels of TNF-α, IL-6, and IL-12p40. Mir-155 deficiency suppresses IL-17 expression, which contributes to the liver IRI development. In our further in vitro study, the results show that the Th17 differentiation is inhibited by SOCS1 overexpression and the promoted M2 macrophage development induced by mir-155 deficiency is abolished by SOCS1 knockdown. In conclusion, mir-155 deficiency attenuates liver IRI through upregulation of SOCS1, and this was associated with promoted M2 macrophage and inhibited Th17 differentiation.

Effects of replacing soybean meal with detoxified Jatropha curcas kernel meal in the diet on growth performance and histopathological parameters of growing pigs

The study was conducted to investigate the effects of replacing soybean meal (SBM) with different levels of detoxified Jatropha curcas kernel meal (DJKM) in growing pig diets on growth performance, haemato-immunological and histopathological parameters. A total of 144 pigs (Duroc×Landrace×Yorkshire) with initial body weight of (20.47±1.44)kg, were randomly allocated to 6 dietary treatments with 6 replications per treatment and 4 pigs per replication for a period of 79 days. The 6 diets (D1, D2, D3, D4, D5 and D6) were formulated using DJKM to replace 0%, 15%, 30%, 45%, 60% and 75% SBM protein and contained phorbol esters (PEs) of 0, 2.75, 5.50, 8.25, 11.00, 13.75mg/kg diets. At d 29, pigs in 3 replicates in D6 treatment were offered D1 diet (expressed as D6+D1) to examine the residual effect of diet D6 in a re-feeding trial. From d 1 to 79 of experimental period, pigs fed diets D4, D5 and D6 had significantly lower average daily gain, average daily feed intake and gain-to-feed ratio than those fed diet D1 (P<0.05). However, the negative effect of diet D6 was disappeared after two weeks of feeding diet D1 in the re-feeding trial. In addition, pigs offered diets D5 and D6 had significantly higher activities of serum alkaline phosphatase, but lower serum alanine transaminase when compared with pigs given diet D1 (P<0.05). Moreover, pathological alterations in liver were observed in pigs fed diets D3, D4, D5 and D6 relative to D1. There results indicate that DJKM could be used to replace SBM protein up to 30% with no detrimental impacts on growth performance, haemato-immunological and pathological parameters in growing pig diet.