Liver Tissue Inflammation Research Articles

The role of primary care physicians in diagnosis and follow-up of patients with chronic viral hepatites B, C and D

By 01.01.2013 in the Republic of Tatarstan there were 23 970 Hepatitis B Surface Antigen carriers and 4546 patients with active Hepatitis B out of 28 516 registered patients who were infected with Hepatitis B. Our experience shows that not only inactive chronic Hepatitis B Surface Antigen carriers, but also patients with different clinical forms of Hepatitis B might be estimated as Hepatitis B Surface Antigen carriers. The feature of chronic viral hepatitides is a discrepancy between the scarce clinical picture and presence (in 15-30% of cases) of active inflammation in liver tissue. Since 2009, patients with positive ELISA test for antibodies to Hepatitis C virus are no longer registered in the national registry of infective diseases of Russian Federation. In the Republic of Tatarstan, such patients with antibodies to Hepatitis C virus were still registered up to 2012 (50 266 patients were registered by 01.01.2012). According to our own data, Hepatitis C viral RNA was found at polymerase chain reaction to Hepatitis C virus in 61% of cases out of 957 patients tested. According to our own data, when using the combined treatment (interferon alpha and ribavirin), lasting virologic response was obtained in 65.7% cases of type 1 genotype of Hepatitis C and almost in 100% of cases of other genotypes out of 163 cases of chronic Hepatitis C, which corresponds to worldwide standards. By 01.01.2013, 295 patients with liver cirrhosis as the result of chronic Hepatitis C were followed up at the consultative and diagnostic center of Republican Clinical Hospital of Infective Diseases with the upward trend. The retrospective analysis of 130 case histories of patients with liver cirrhosis revealed that Hepatitis C was firstly diagnosed at the stage of established liver cirrhosis in 57.7% of cases. Timely diagnosis and periodic medical examination in cases of chronic Hepatitis C allows to implement the individual, personalized approach in such patients and to choose the adequate treatment and follow-up.

Myeloid Cell-specific Disruption of Period1 and Period2 Exacerbates Diet-induced Inflammation and Insulin Resistance

The circadian clockworks gate macrophage inflammatory responses. Given the association between clock dysregulation and metabolic disorders, we conducted experiments to determine the extent to which over-nutrition modulates macrophage clock function and whether macrophage circadian dysregulation is a key factor linking over-nutrition to macrophage proinflammatory activation, adipose tissue inflammation, and systemic insulin resistance. Our results demonstrate that 1) macrophages from high fat diet-fed mice are marked by dysregulation of the molecular clockworks in conjunction with increased proinflammatory activation, 2) global disruption of the clock genes Period1 (Per1) and Per2 recapitulates this amplified macrophage proinflammatory activation, 3) adoptive transfer of Per1/2-disrupted bone marrow cells into wild-type mice potentiates high fat diet-induced adipose and liver tissue inflammation and systemic insulin resistance, and 4) Per1/2-disrupted macrophages similarly exacerbate inflammatory responses and decrease insulin sensitivity in co-cultured adipocytes in vitro. Furthermore, PPARγ levels are decreased in Per1/2-disrupted macrophages and PPARγ2 overexpression ameliorates Per1/2 disruption-associated macrophage proinflammatory activation, suggesting that this transcription factor may link the molecular clockworks to signaling pathways regulating macrophage polarization. Thus, macrophage circadian clock dysregulation is a key process in the physiological cascade by which diet-induced obesity triggers macrophage proinflammatory activation, adipose tissue inflammation, and insulin resistance.

Reversion from a high‐fat diet to a prenatal normal diet could delay the incidence of insulin resistance induced by postnatal high fat diet in female offspring (1072.2)

Many studies have focused on the adverse effects of prenatal high fat diet (HFD) on offspring development, but few studies are aimed on evaluating how prenatal reversion to regular diet (ND) in obese subjects could affect the offspring high fat diet‐induced obesity (DIO). The female obese mice either continued a high fat diet during pregnancy till the end of lactation (HFD group), or switched from HFD to ND for four weeks and continued on ND during pregnancy till the end of lactation (HFD‐ND group). Offspring from both groups were subjected on HFD for 9 weeks after weaning. By the end of the week 9, we found that prenatal HFD‐ND differentially affects the male and female offspring with a postnatal HFD. This is evidenced by slower weight gain, lower weight of the liver, the subcutaneous and the visceral fat in HFD‐ND female offspring compared to the HFD female offspring, but not in HFD‐ND male offspring vs. HFD male offspring. Only HFD‐ND female offspring failed to develop insulin resistance after nine weeks postnatal HFD. This result was not caused by the differential consumption of male and female offspring via evaluating the food consumption. We further found that the female HFD offspring developed inflammation in liver tissue marked by increased macrophage infiltration comparing to the HFD‐ND female offspring. Our results demonstrated that maternal reversion from a HFD to a ND could affect the onset of DIO in female offspring.Grant Funding Source: Supported by the University of North Dakota faculty seed money

Dual role of B7 costimulation in obesity-related non-alcoholic steatohepatitis (NASH) and metabolic dysregulation

The inflammatory state present in obesity contributes to obesity-related metabolic dysregulation, including non-alcoholic steatohepatitis (NASH) and insulin resistance. Intercellular interactions between immune cells or between immune cells and hepatic parenchymal cells contribute to the exacerbation of liver inflammation and steatosis in obesity. The co-stimulatory molecules B7.1 and B7.2 are important regulators of cell-cell interactions in several immune processes; however, the role of B7 co-stimulation in obesity-related liver inflammation is still unknown. Here, diet-induced obesity (DIO) studies in mice with genetic inactivation of both B7.1 and B7.2 (Dko) revealed aggravated obesity-related metabolic dysregulation, glucose intolerance and enhanced progression to steatohepatitis due to B7.1/B7.2-double deficiency. The metabolic phenotype of B7.1/B7.2-double deficiency upon DIO was accompanied by increased hepatic and adipose tissue inflammation, associated with largely reduced numbers of regulatory T cells (Tregs) in these organs. In order to assess the role of B7co-stimulation in DIO in a non-Treg lacking environment, we performed antibody-mediated inhibition of B7 molecules in wild-type mice in DIO. Antibody-blockade of both B7.1 and B7.2, but not of either one alone, improved the metabolic phenotype of DIO-mice, which was linked to amelioration of hepatic steatosis and reduced inflammation in liver and adipose tissue. Our study demonstrates a dual role of B7 co-stimulation in the course of obesity-related sequelae, particularly NASH. The genetic inactivation of B7.1/B7.2 deteriorates obesity-related liver steatosis and metabolic dysregulation likely due to the intrinsic absence of Tregs in these mice rendering Dko mice a novel murine model of NASH in future studies. In contrast, inhibition of B7 co-stimulation under conditions, where Tregs are present, may provide a novel therapeutic approach for obesity-related metabolic dysregulation and especially NASH.

Caffeinated and decaffeinated instant coffee consumption partially reverses high-fat diet-induced metabolic alterations in mice

Epidemiological data has associated coffee consumption with a lower prevalence of type 2 diabetes, metabolic syndrome and chronic liver disease. However, the mechanisms and coffee substances responsible for these effects remain unclear. In the present study, mice received caffeinated or decaffeinated instant coffee ad libitum during a two week period after inducing obesity by introducing a high-fat diet over a 10week period. Body weight, glucose homeostasis, and liver and visceral adipose tissue inflammation were assessed. In addition, AKT signaling, the fatty acid profile and liver histopathological analyses were performed. Ingestion of caffeinated or decaffeinated coffee for 2weeks resulted in reductions in glucose and insulin blood levels, and insulin tolerance was improved without reductions in final body weight or adiposity. Only caffeinated coffee modified the adipokine profile in visceral adipose tissue, resulting in a restoration of adiponectin levels. However, deleterious liver alterations, which manifested as reductions in steatosis, inhibition of iNOS expression and restoration of insulin inducing-AKT phosphorylation, were reversed by the ingestion of both caffeinated and decaffeinated coffee. Serum AST and ALT levels were also improved in mice after coffee ingestion, while the fatty acid profile in the liver and the cytokine profiles in adipose and liver tissues were not altered by coffee ingestion. Thus, instant coffee consumption reverses obesity-induced alterations in mice, and our results corroborate those of epidemiological studies that associated coffee consumption with a lower prevalence of obesity-related pathologies.

Correlation of hepatocyte expression of hepatitis B viral core antigen and the clinicopathological characters in chronic hepatitis B patients

To investigate the relationship between the expression of hepatitis B virus (HBV) core antigen and viral replication and liver tissue inflammation damage in chronic hepatitis B (CHB) patients, and to analyze the relationship of core antigen expression differences with clinical and pathological features in e antigen-negative and e antigen-positive CHB patients. Sixty-three treatment-naive patients diagnosed with CHB who underwent liver biopsy were included in this retrospective analysis. Liver pathology was assessed, and the karyotype, pulp type, and pulp karyotype were determined. Core and e antigen expression was quantitatively determined by automated immunoassay. Blood samples were used to determine the amount of peripheral lymphocytes or monocytes and HBV DNA load. Results The median titer of HBV DNA was significantly higher in the CHB patients with e antigen positivity (n = 48) than those with e antigen negativity (n = 15) (5.4 * 106 copies/ml vs. 5.4 * 104 copies/ml, P = 0.003). The core antigen positive expression rate was significantly higher in the e antigen-positive CHB patients than in the e antigen-negative CHB patients (80.33% vs. 53.33%, P = 0.042). For the e antigen-positive CHB patients, the HBV DNA titer in karyotype core antigen cases was higher than that in the pulp karyotype mixed-type cases (P = 0.008) and in the negative cases (P = 0.013); in addition, the karyotype patients showed higher titer than the plasma patients (P = 0.019). Also for the e antigen-positive CHB patients, the HBV DNA titer was positively correlated with the rank level of pulp karyotype in core antigen expression (r = 0.589, P = 0.003) but negatively correlated with lobular inflammation, interface inflammation, and fibrosis level (r = -0.552, P = 0.000; r = -0.381, P = 0.008; r = -0.555, P = 0.000); in addition, the level of peripheral blood lymphocytes was negatively correlated with lobular inflammation and fibrosis level (r = -0.361, P = 0.012; r = -0.356, P = 0.013). For the e antigen-negative CHB patients, the level of peripheral blood lymphocytes was negatively correlated with lobular inflammation and interface inflammation (r = -0.702, P = 0.004; r = -0.578, P = 0.024), while the level of peripheral blood mononuclear cells was negatively correlated with lobular inflammation, interface inflammation, and fibrosis level (r = -0.682, P = 0.005; r = -0.620, P = 0.014; r = -0.527, P = 0.044); in addition, age positively correlated with interface inflammation (r = 0.690, P = 0.004). The pulp karyotype mixed-type of core antigen expression may reflect the level of HBV replication. Negative expression of core antigen may be associated with variation in pre-C or C zone. The monocyte-macrophage system may be involved in the pathogenesis of e antigen-negative CHB, while the mechanism of immune escape may play an important role in increasing HBV DNA titer in an e-antigen-negative CHB condition.

Hepatoprotective effect of aqueous and pure extracts of raisin in alloxan-induced diabetic rats

Abstract .Thepresentstudywasconductedtoevaluatethepossibleantioxidanteffectsofraisinaqueousandpurextractof“Karkni”on liver histology, antioxidant enzymes activities and lipid peroxide levels in liver of diabetic and normal rats. 125mg/kg of pureand aqueous extract of “Karkni” were orally administered daily to alloxan-diabetic rats for 4 weeks. Alloxan- induced diabetic ratsshowed significant increases in the levels of total protein and malondialdehyde (MDA), glutathione peroxidase (GPx), catalase(CAT) and reduced glutathione (GSH) levels were significantly decreased compared to normal rats ( p <0.05). Also, no significanteffectsonglutathioneperoxidase(GPx)activityindiabeticratswereobserved.Thechangesoftheaboveparameterstotheirnormallevels after 4 weeks of treatment were observed mainly with the pure extract. The results suggesting that pure extract suppressesthe liver tissue inflammation attenuate lipid peroxidation and can be helpful in reducing liver damage caused by alloxan-induceddiabetes.Keywords: “Karkni”, alloxan, liver, antioxidant enzymes, MDA

Infection by Brazilian and Dutch swine hepatitis E virus strains induces haematological changes in Macaca fascicularis.

BackgroundHepatitis E virus (HEV) has been described as an emerging pathogen in Brazil and seems to be widely disseminated among swine herds. An autochthonous human case of acute hepatitis E was recently reported. To obtain a better understanding of the phenotypic profiles of both human and swine HEV strains, a experimental study was conducted using the animal model, Macaca fascicularis.MethodsSix cynomolgus monkeys (Macaca fascicularis) were inoculated intravenously with swine HEV genotype 3 that was isolated from naturally and experimentally infected pigs in Brazil and the Netherlands. Two other monkeys were inoculated with HEV genotype 3 that was recovered from Brazilian and Argentinean patients with locally acquired acute and fulminant hepatitis E. The haematological, biochemical, and virological parameters of all animals were monitored for 67 days.ResultsSubclinical hepatitis was observed in all monkeys after inoculation with HEV genotype 3 that was recovered from the infected swine and human patients. HEV RNA was detected in the serum and/or faeces of 6 out of the 8 cynomolgus monkeys between 5 and 53 days after inoculation. The mild inflammation of liver tissues and elevations of discrete liver enzymes were observed. Seroconversions to anti-HEV IgM and/or IgG were detected in 7 animals. Reactivities to anti-HEV IgA were also detected in the salivary samples of 3 animals. Interestingly, all of the infected monkeys showed severe lymphopenia and a trend toward monocytosis, which coincided with elevations in alanine aminotransferase and antibody titres.ConclusionsThe ability of HEV to cross the species barrier was confirmed for both the swine (Brazilian and Dutch) and human (Argentinean) strains, thus reinforcing the zoonotic risk of hepatitis E in South America. Cynomolgus monkeys that were infected with HEV genotype 3 developed subclinical hepatitis that was associated with haematological changes. Haematological approaches should be considered in future studies of HEV infection.

Interleukin-4 deficiency protects mice from acetaminophen-induced liver injury and inflammation by prevention of glutathione depletion

Interleukin-4 (IL-4) is a multifunctional cytokine involved in many diseases such as autoimmune hepatitis and idiosyncratic drug reactions. However, its role in acetaminophen (APAP)-induced liver injury remains unclear. Our objective was to evaluate the contribution of IL-4 to the pathogenesis of APAP-induced liver injury. Balb/C (WT) and IL-4 knockout (IL-4(-/-)) mice were orally overdosed with APAP. After 24 h, survival percentage, biochemical and morphological markers of liver injury, and tissue inflammation were assessed. IL-4(-/-) mice were protected from APAP toxicity. Intravital confocal microscopy, tissue histology and serum ALT levels showed significantly less liver injury and inflammation than in the WT group, which may explain the increased survival rate of IL-4(-/-) mice. In addition, IL-4(-/-) mice had decreased production of tumor necrosis factor α, CXCL1 and interleukin-1β in the liver, but not in a remote site such as the lungs. Hepatic macrophage activation was markedly reduced in IL-4-deficient mice. In addition, glutathione depletion-a primary cause of APAP-mediated injury-was significantly attenuated in IL-4(-/-) mice. Taken together, our data demonstrate that IL-4(-/-) mice are protected from APAP-induced liver injury due to reduced depletion of glutathione, which prevented liver damage and tissue inflammation.

Paraoxonase-1 status in patients with hereditary hemochromatosis

Hereditary hemochromatosis (HH) is characterized by accumulation of iron, oxidative stress, inflammation, and fibrogenesis in liver tissue. In this setting, research on the protection afforded by intracellular antioxidants is of clinical relevance. Paraoxonase-1 (PON1) is an enzyme that degrades lipid peroxides. This study investigates the alterations in serum PON1 status, PON1 gene polymorphisms, and PON1 hepatic expression in patients with HH. We performed a case-control study in 77 patients with HH (80.5% men, 22-70 years of age) and 408 healthy individuals (43.1% men, 26-74 years of age). Serum PON1 activities against different substrates and PON1192 and PON155 polymorphisms were analyzed. PON1 protein expression was investigated in 20 liver biopsies. HH patients had significantly lower serum PON1 activity, which was inversely correlated with ferritin (marker of iron stores) and serum 8-isoprostane concentrations (index of oxidative stress). PON1 protein expression in liver tissue was higher in patients and showed stronger staining in hepatocytes surrounding the areas of inflammation. Our study provides preliminary evidence that PON1 may play a role in protecting against iron-induced oxidative stress in hereditary hemochromatosis.

Dietary eicosapentaenoic acid supplementation accentuates hepatic triglyceride accumulation in mice with impaired fatty acid oxidation capacity

Reduced mitochondrial fatty acid (FA) β-oxidation can cause accumulation of triglyceride in liver, while intake of eicosapentaenoic acid (EPA) has been recommended as a promising novel therapy to decrease hepatic triglyceride content. However, reduced mitochondrial FA β-oxidation also facilitates accumulation of EPA. To investigate the interplay between EPA administration, mitochondrial activity and hepatic triglyceride accumulation, we investigated the effects of EPA administration to carnitine-deficient mice with impaired mitochondrial FA β-oxidation. C57BL/6J mice received a high-fat diet supplemented or not with 3% EPA in the presence or absence of 500mgmildronate/kg/day for 10days. Liver mitochondrial and peroxisomal oxidation, lipid classes and FA composition were determined. Histological staining was performed and mRNA level of genes related to lipid metabolism and inflammation in liver and adipose tissue was determined. Levels of pro-inflammatory eicosanoids and cytokines were measured in plasma. The results showed that mildronate treatment decreased hepatic carnitine concentration and mitochondrial FA β-oxidation and induced severe triglyceride accumulation accompanied by elevated systemic inflammation. Surprisingly, inclusion of EPA in the diet exacerbated the mildronate-induced triglyceride accumulation. This was accompanied by a considerable increase of EPA accumulation while decreased total n-3/n-6 ratio in liver. However, inclusion of EPA in the diet attenuated the mildronate-induced mRNA expression of inflammatory genes in adipose tissue. Taken together, dietary supplementation with EPA exacerbated the triglyceride accumulation induced by impaired mitochondrial FA β-oxidation. Thus, further thorough evaluation of the potential risk of EPA supplementation as a therapy for NAFLD associated with impaired mitochondrial FA oxidation is warranted.

Effects of Acute Intracranial Hypertension on Extracerebral Organs: A Randomized Experimental Study in Pigs

The study was conducted to determine the effects of isolated acute intracranial hypertension (AICH) on extracerebral organs. A total of 14 mechanically ventilated pigs were randomized to two groups of seven each: (1) control and (2) AICH. AICH was induced by inflating an intracranial balloon catheter. The inflation volume was adjusted to keep intracranial pressure between 30 and 40 cm H2O. Hemodynamics, gas-exchange, and global oxygen delivery parameters were observed over a 4-hour period. At the end of the 4-hour period, tissue samples of heart, lungs, liver, and kidneys were collected and histologically graded for inflammation, edema, and cell damage (necrosis) using semiquantitative scores. Animals with AICH had increased heart rate and cardiac output, and higher scores for inflammation, edema, and necrosis in heart, lung, kidney, and liver tissues (all p < 0.05). Peripheral and mixed-venous oxygen saturations were unaffected. Isolated AICH induces injury to multiple extracerebral organs, even in the absence of hypoperfusion or hypoxemia.

Calorie Restriction with a High-Fat Diet Effectively Attenuated Inflammatory Response and Oxidative Stress-Related Markers in Obese Tissues of the High Diet Fed Rats

Obesity characterized by increased mass of adipose tissue leads to systemic inflammation. Calorie restriction (CR) improves parameters associated with immune response and antioxidant defense. We hypothesized that CR with a high fat diet (HFCR) regulates local and systemic inflammation and oxidative stress damage in a high fat diet induced obesity (HF group). We investigated effect of HFCR on inflammation and oxidative stress-related markers in liver and adipose tissues as well as adipokines in plasma. HFCR lowered liver triglyceride levels, total cholesterol levels, and the plasma leptin/adiponectin ratio to normal levels and improved glucose tolerance. HFCR also improved fatty liver and normalized adipocyte size and morphology. HFCR reduced lipid peroxidation and decreased the expression levels of inducible nitric oxide synthetase, cyclooxygenase-2, NF-E2-related factor, and heme oxygenase-1 in the liver. Moreover, HFCR suppressed the expression levels of C- reactive protein and manganese superoxide dismutase in the adipose tissue in the HF group. These results suggest that HFCR may have beneficial effects on inflammation and oxidative stress as well as lipid profiles in the HF diet induced obesity. Moreover, HFCR may be a good way to increase compliance in obese patients and to prevent obesity induced complications without changes in dietary pattern.

Comparative analysis of the human hepatic and adipose tissue transcriptomes during LPS-induced inflammation leads to the identification of differential biological pathways and candidate biomarkers

BackgroundInsulin resistance (IR) is accompanied by chronic low grade systemic inflammation, obesity, and deregulation of total body energy homeostasis. We induced inflammation in adipose and liver tissues in vitro in order to mimic inflammation in vivo with the aim to identify tissue-specific processes implicated in IR and to find biomarkers indicative for tissue-specific IR.MethodsHuman adipose and liver tissues were cultured in the absence or presence of LPS and DNA Microarray Technology was applied for their transcriptome analysis. Gene Ontology (GO), gene functional analysis, and prediction of genes encoding for secretome were performed using publicly available bioinformatics tools (DAVID, STRING, SecretomeP). The transcriptome data were validated by proteomics analysis of the inflamed adipose tissue secretome.ResultsLPS treatment significantly affected 667 and 483 genes in adipose and liver tissues respectively. The GO analysis revealed that during inflammation adipose tissue, compared to liver tissue, had more significantly upregulated genes, GO terms, and functional clusters related to inflammation and angiogenesis. The secretome prediction led to identification of 399 and 236 genes in adipose and liver tissue respectively. The secretomes of both tissues shared 66 genes and the remaining genes were the differential candidate biomarkers indicative for inflamed adipose or liver tissue. The transcriptome data of the inflamed adipose tissue secretome showed excellent correlation with the proteomics data.ConclusionsThe higher number of altered proinflammatory genes, GO processes, and genes encoding for secretome during inflammation in adipose tissue compared to liver tissue, suggests that adipose tissue is the major organ contributing to the development of systemic inflammation observed in IR. The identified tissue-specific functional clusters and biomarkers might be used in a strategy for the development of tissue-targeted treatment of insulin resistance in patients.

Expression of COX-2 and PPARg in the livers of patients with acute on chronic HBV-related liver failure and their relationship with clinic parameters

To study the expressions of cyclooxygenase-2 (COX-2) and Peroxisome proliferator-activated receptor gamma (PPARg) in liver of patients with hepatitis B virus (HBV) related acute-on-chronic liver failure (ACLF) and their correlation with clinical parameters. 35 patients with ACLF, 35 patients with HBV related chronic liver failure (CLF), 27 patients with chronic hepatitis B(CHB) and 15 normal control were enrolled to study the expressions of COX-2 and PPARg in the liver tissues by immunohistochemical staining, and to analyze the correlation of the COX-2 and PPARg levels in liver tissues with clinical parameters. COX-2 was distinctly expressed in the cytoplasm of the hepatocytes, but PPARg was mostly expressed in the nuclei of the hepatocytes and also could be seen in the cytoplasm. The expressions of COX-2 in the liver of ACLF, CLF and CHB groups increased significantly as compared with NC group (z = -5.18, -4.50, -5.32, P is less than 0.01). The levels of COX-2 in ACLF livers also increased evidently as compared with CLF groups (z = -1.98, P is less than 0.05). The expression levels of PPARg in ACLF liver tissues were much higher than the other three groups, and statistical significances existed between ACLF group and the other two groups (CLF, NC groups) (z = -2.62, -4.28, P is less than 0.01). In ACLF group, the expression of COX-2 correlated with MELD score (r = 0.337, P is less than 0.05) and the expression of PPARg correlated with HBV DNA load (r = 0.348, P is less than 0.05). Clinical data showed that the levels of AST, TBil, CHOL, PT, INR, FIB and MELD score in ACLF group were significantly different from that in CLF, CHB and NC groups. COX-2 expressed in liver may be a marker to reflect the degree of inflammation and injury of liver tissue. The PPARg expression of liver could be increased during chronic HBV infection and may be a protective mechanism against liver injury.

Effects of Chinese herbal medicine Fuzheng Huayu recipe and its components against hepatocyte apoptosis in mice with hepatic injury

To investigate the effects and characteristics of Fuzheng Huayu recipe, a compound Chinese herbal medicine, and its decomposed therapies against hepatocyte apoptosis in mice with hepatic injury. A total of 50 male BALB/c mice were randomly divided into 5 groups: control group, untreated group, Fuzheng Huayu recipe group, Fuzheng recipe group and Huayu recipe group. Hepatocyte apoptosis in mice was induced by intraperitoneal injection of lipopolysaccharide (10 μg/kg) and galactosamine (900 mg/kg). The mice in drug-treated groups were administered with Fuzheng Huayu recipe, Fuzheng recipe and Huayu recipe by garbage respectively 3 days before injection of lipopolysaccharide and galactosamine. The mice were sacrificed 6 hours after the administration of lipopolysaccharide and galactosamine. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and superoxide dismutase (SOD) and content of malondialdehyde (MDA) were examined by colorimetric method. Inflammation and necrosis in liver tissue were observed with hematoxylin-eosin staining. Hepatocyte apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. Protein expression of tumor necrosis factor receptor type I (TNFR1) was analyzed with Western blotting. Expression of tumor necrosis factor-α (TNF-α) mRNA was analyzed with real-time fluorescent quantitative polymerase chain reaction. Compared with the untreated group, Fuzheng Huayu recipe, Fuzheng recipe and Huayu recipe attenuated hepatocyte apoptosis, decreased the serum ALT and AST activities and MDA content, and improved the SOD activity in liver tissues. Of the three groups, the effect of Fuzheng Huayu recipe group was the best, Fuzheng recipe group was better than Huayu recipe group. Compared with the untreated group, all drugs exerted good effects in decreasing TNF-α mRNA expression, and Fuzheng Huyu recipe was better than the others. The expression of TNFR1 protein in the untreated group was increased with the occurrence of hepatocyte apoptosis, however, only Fuzheng Huayu recipe decreased the elevated TNFR1 protein expression. Fuzheng recipe exerts a good effect in attenuating hepatocyte apoptosis in vivo, which has synergistic effects with Huayu recipe in attenuating hepatocyte apoptosis, and the potential mechanism is partially due to their different effects in alleviating oxidative stress in liver and down-regulating the expression of TNFR1 protein.

RETRACTED: Continuous Hemodiafiltration Therapy Ameliorates LPS-Induced Systemic Inflammation in a Rat Model

Systemic inflammation can result in multiple organ dysfunction syndrome, a potentially life-threatening condition. Some reports suggest that continuous hemodiafiltration can effectively remove proinflammatory cytokines from circulation during systemic inflammation. In the present study, we investigated whether continuous hemodiafiltration therapy could prevent LPS-induced systemic inflammation and improve survival in a rat model. Male Wistar rats were injected with lipopolysaccharide (LPS; 7.5 mg/kg body weight), and 6 h later were treated with either single-pass hemofiltration (C group), continuous hemofiltration (CHF group), continuous hemodiafiltration (CHDF group), or mock filtration (Control group). We performed histologic examinations of lung and liver tissues, determined serum cytokine levels, and survival rates for each treatment group, and compared cytokine removal between CHF and CHDF therapies. Histologic examination revealed significant reduction in inflammation in lung and liver tissues harvested 24 h after CHDF compared with the Control group. Likewise, LPS-induced serum TNF-α and IL-6 levels decreased with continuous hemodiafiltration along with a significant improvement in survival. After 30 min of treatment, both CHF and CHDF removed significant amounts of TNF-α and IL-6 from the blood. However, serum cytokine levels measured before and after filtration were not significantly different. Continuous hemodiafiltration therapy lowered inflammatory cytokines and increased survival rates in a rat model of systemic inflammation. Therefore, continuous hemodiafiltration may be a potential therapy for use against various systemic inflammatory diseases.

Impaired transactivation of the human CYP2J2 arachidonic acid epoxygenase gene in HepG2 cells subjected to nitrative stress

Human cytochrome P450 2J2 (CYP2J2) generates epoxyfatty acids that modulate cellular apoptosis and proliferation. CYP2J2 regulation has not been intensively studied but induction of the activator protein-1 (AP-1) subunit c-fos mediates CYP2J2 down-regulation in hypoxia, a component of ischaemic injury. Decreased CYP2J2 expression may contribute to tissue injury. HepG2 cells were treated with sodium nitroprusside (SNP) to induce nitrative stress, which has been associated with inflammation and infection in liver and other tissues. CYP2J2 protein and mRNA expression were evaluated by immunoblotting and real-time PCR respectively. The role of mitogen-activated protein (MAP) kinases in CYP2J2 dysregulation was assessed using specific inhibitors and dominant negative MAP kinase expression plasmids. CYP2J2-luciferase reporter constructs and electromobility shift assays (EMSAs) were used to identify SNP-regulated regions in the CYP2J2 gene. Cytochrome P450 2J2 was down-regulated by SNP while the AP-1 proteins c-jun and c-fos were up-regulated; inhibition of p38 and ERK MAP kinases normalized their expression. The gene elements at -105/-95 and -56/-63 were required for the down-regulation of CYP2J2 induced by nitrative stress. p38 and ERK MAP kinases transduce stress stimuli that down-regulate CYP2J2. Targeting these kinases may prevent the loss of CYP2J2 and epoxy-fatty acids that protect cells against deleterious stresses.

Effects of Qushi Huayu Decoction in prevention and treatment of fatty liver in rats based on adiponection-free fatty acid pathway

To explore the effects of Qushi Huayu Decoction (QSHYD), a compound traditional Chinese herbal medicine, in prevention and treatment of non-alcoholic fatty liver disease (NAFLD) in rats. Forty Wistar male rats were used to establish the NAFLD model by subcutaneous injection of carbon tetrachloride (CCl(4)) for 4 weeks (twice weekly) along with high-fat and low-protein diet for 2 weeks. After two-week administration, the rats were randomly divided into four groups: untreated group, high-dose QSHYD group, medium-dose QSHYD group and low-dose QSHYD group. Another six rats were used as normal control. After 2-week treatment, the following indexes were detected: (1) liver pathology; (2) contents of serum adiponectin (ADP) and liver triglyceride (TG); (3) concentrations of liver FFA, adiponectin receptor 2 (AdipoR2), malonyl-coenzyme A (malony1-CoA), AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACCase), fatty acid synthase (FAS) and carnitine palmitoyl transferase-1 (CPT-1). Compared with the normal group, there were physiological changes associated with hepatic steatosis and inflammation in liver tissues in the untreated group as observed by oil red O staining and HE staining. The TG, FFA, malony1-CoA, FAS, and ACCase concentrations in liver tissues in the untreated group were elevated significantly. While the contents of ADP in serum and AdipoR2, CPT-1 and AMPK in liver tissues in the untreated group were decreased markedly. The pathological damages in each QSHYD-treated group were significantly less than those in the untreated group. The TG and FFA contents in liver tissues in each QSHYD-treated group were significantly decreased. The FAS, ACCase and malonyl-CoA concentrations in liver tissues of the high QSHYD-treated group were reduced markedly as compared with the untreated group. High- and medium-dose of QSHYD could significantly increase ADP content in serum and AMPK, CPT-1 and AdipoR2 contents in liver tissues. QSHYD can affect the ADP-FFA pathway by increasing the content of serum ADP, which may be one of its important mechanisms in preventing and treating NAFLD in rats.

Level and clinical significance of soluble CD40 in patients with chronic hepatitis B

To understand the level and clinical significance of soluble CD40 in patients with chronic hepatitis B. Detecting the concentration of sCD40 from 176 cases with chronic hepatitis B by ELISA and analyzing its relationship with different grades of inflammation and necrosis in liver tissue. sCD40 from patients with chronic hepatitis B was significantly higher than those from healthy. And that the concentration of sCD40 was positive correlation with severe clinical disease and liver inflammation and necrosis. In patients whose ALT lower than 80 IU/L and sCD40 higher than 80 pg/ml, it showed that 65.85% cases have high grade of liver inflammation and necrosis, which was significantly higher than patients with sCD40 lower than 80 pg/ ml. The concentration of sCD40 is positively related with the grade of liver inflammation and necrosis. This information could help us to evaluate the status of chronic hepatitis B as an immunological index.