Knodell Scoring System Research Articles

Peripheral immune cell profile and activation status in chronic HCV mono- and HCV/HIV co-infected patients

Background: Due to similar routes of viral transmission, chronic HCV and HIV co-infection is frequent. Since it has been reported that both viruses perturbs immune homeostasis, our aim was to evaluate the peripheral immune cell moiety in HCV mono- and HCV/HIV co-infected patients to determine if co-infection alters HCV immune profile. Methods & Materials: Liver biopsies (47 = HCV treatment naïve, 13 = HCV/HIV only HAART) and concomitant blood samples were analysed. In peripheral blood: 1) frequency of total T cells(CD3 + ), T helper(Th, CD3 + CD4 + ), cytotoxic T lymphocytes(CTL, CD3 + CD8 + ) and Natural Killer cells(NK, CD3-CD56 + CD16 + ); 2) CTL and Th differentiation status [naïve (CD27 + CD45RA + ), central-memory (CD27 + CD45RA-), effector-memory(CD27-CD45RA-), effector(CD27-CD45RA + ) and activated (DR + )]; 3) frequencies of Th17(CD4 + IL17A + ), Th1(CD4 + IFNg + ) and Treg(CD4 + CD25hiFoxp3 + ) and 4) NK and CTL degranulation activity (CD107a) were evaluated by FACS. Non-infected donors (n = 33) were included. Inflammatory activity and fibrosis were assessed in liver biopsies using the modified Knodell scoring system and METAVIR. Results: While HCV patients did not display differences in lymphocyte frequencies compared to donors, co-infected patients showed the lowest Th(donor vs HCV/HIV p < 0.001, HCV vs HCV/HIV p < 0.001), the highest CTLs(donor vs HCV/HIV p < 0.001, HCV vs HCV/HIV p < 0.001) and the lowest NK(donor vs HCV/HIV p < 0.01, HCV vs HCV/HIV p < 0.05) frequencies. Absolute numbers confirmed these results. Moreover, only Th17 absolute numbers were diminished in co-infection (donor vs HCV/HIV p < 0.05). Regarding T cell differentiation status in HCV patients compared to donors: Th cells showed low naïve (p < 0.01) but high effector memory (p < 0.05) and activated cells (p < 0.01) frequencies; CTL showed low naïve (p < 0.01) but high activated cell (p < 0.01) frequencies. Although, HCV/HIV displayed a similar profile to HCV, Th and CTL activation was higher in HCV/HIV than HCV (p < 0.001). Degranulation activity of CTL and NK cells showed no differences between groups. Concerning to liver damage, any of the evaluated immune cell parameters displayed association with fibrosis or hepatitis severity. Conclusion: Although in chronic HCV infection the lymphocyte frequency was not modified, an altered T cell differentiation profile was evident. The co-infected condition distorted lymphocyte frequencies and enhanced T cell activation. This deteriorated immune profile may condition T cell response against antigens, including vaccines.

The interplay between intrahepatic lymphocyte populations and Hepatitis B virus antigens related to liver damage in chronic hepatitis

Background: Chronic hepatitis B virus (HBV) infection is still a global health problem. Even though it is well known that adaptative immune response plays an important role in the pathogenesis of chronic infection, the impact of each immune cell population in liver damage and viral surveillance is still a matter of debate. Our aim was to elucidate the interplay between the immune response and viral activity in the context of liver damage. Methods & Materials: Immunostaining was performed in 23 liver biopsies from CHB treatment naïve patients (40% HBeAg-positive) to:1) characterize liver infiltrate [Th (CD4+), Th1 (Tbet+), Th17 (IL-17A+), Treg (Foxp3+), and CTL (CD8+)] [portal quantification: immunostained/total lymphocytes; lobular quantification: immunostained lymphocytes in 10 fields; (400x)], 2) evaluate the Hepatitis B surface antigen (HBsAg) expression (presence was determined). Inflammatory activity and fibrosis were assessed using the modified Knodell scoring system (Histological Activity Index, HAI) and METAVIR. Results: All studied populations were observed in portal/periportal infiltrates with predominance of Th {Th [0.65 (0.33-0.77)] > CTL [0.54 (0.16-0.74)] > Treg [0.09 (0.00-0.25)] > Th1 [0.07 (0-0.20)] > Th17 [0.05 (0-0.20)]}. However, only CTL were present in the intralobular area [2.77 (0.01-0.25)]. Concerning liver damage, Th17 frequency was increased in cases with more severe fibrosis (p = 0.04), while Th in cases with severe hepatitis (p = 0.02) mostly due to Treg (p = 0.02). The frequency of intralobular CTL also depicted association with severe hepatitis (p = 0.02). Regarding HBV antigen profile, only 62% of cases showed presence of HBsAg, being its absence associated with a higher number of Treg (p = 0.02) and severe hepatitis (p = 0.01). Moreover, HBe Ag-positive cases exhibited higher Treg and Th frequencies (p = 0.0002 and p = 0.037; respectively). Conclusion: Although Treg population was highly represented in the infiltrate, they were not related to liver damage. In contrast, intrahepatic Th17 mediated fibrosis severity. The presence of CTL in the intralobular area may indicate a relationship between them and hepatocytes denoting their contribution to hepatitis severity. Finally, our results suggest that the presence of a regulatory microenvironment enhances viral replication. This hypothesis is supported by the fact that a higher Treg frequency was observed in the context of absence HBsAg as well as among HBeAg-positive cases.

Acoustic radiation force impulse quantification of spleen elasticity for assessing liver fibrosis.

The purpose of this study is to assess the correlation between liver fibrosis and spleen stiffness measured by ARFI in patients with chronic viral hepatitis (B or C) and to explore the possible complementary role of spleen and liver ARFI in grading liver fibrosis. 84 subjects (51 patients, 33 healthy volunteers) were enrolled. ARFI of the spleen and the liver was performed. Patients subsequently underwent liver biopsy for grading liver fibrosis according to Knodell scoring system. Multivariate logistic regression and decision tree analysis were adopted to test the relationship between spleen and liver stiffness (independent variables) and liver fibrosis (F1< vs. ≥F3). Leave-One-Out Cross-Validation was used for validating the predictive classification models. Area under the ROC curve (AUROCC) was used as accuracy metric. Spleen ARFI was able to discriminate early (F1) from severe (≥F3) liver fibrosis with an optimal cut-off of 3.05 m/s: AUROCC 0.807, cross-validated AUROCC 0.614. Liver ARFI was superior to spleen ARFI, using a cut-off of 2.11 m/s: AUROCC 0.879, cross-validated AUROCC 0.672. Neither spleen nor liver ARFI was able to differentiate healthy volunteers from F1 patients. Odds ratios derived from logistic regression were 23.1 and 9.9 for liver and spleen ARFI, respectively; resulting AUROCC was 0.905 (cross-validated 0.848). A decision tree considering the sequential use of liver and spleen ARFI with cut-off of 2.14 and 3.39 m/s, respectively, resulted in AUROCC of 0.903 (cross-validated 0.7). Spleen ARFI has the potential to discriminate early from severe liver fibrosis. Spleen and liver ARFI, when combined, show a better discriminative power than liver ARFI alone.

The Clinical Significance of Serum Apoptotic Cytokeratin 18 Neoepitope M30 (CK-18 M30) and Matrix Metalloproteinase 2 (MMP-2) Levels in Chronic Hepatitis B Patients with Cirrhosis

BackgroundSerum apoptotic cytokeratine 18 neoepitope M30 (CK-18 M30) and matrix metalloproteinase 2 (MMP-2) have been popular markers for detecting liver fibrosis in recent years. CK-18 is a major intermediate filament protein in liver cells and one of the most prominent substrates of caspases during hepatocyte apoptosis. MMP-2 plays an important role in tissue remodeling and repairing processes during physiological and pathological states.ObjectivesThe objective of this study was to investigate the significance of CK-18 M30 and MMP-2 levels for clinical use in patients with chronic hepatitis B (CHB), as well as their sensitivity in determining cirrhotic patients.Patients and MethodsThis study included 189 CHB patients and 51 healthy controls. A modified Knodell scoring system was used to determine the fibrosis level in chronic hepatitis B patients. CK-18 M30 levels were determined with an M30-Apoptosense ELISA assay. MMP-2 levels were determined with the ELISA assay.ResultsThe study group consisted of 132 (69.8%) males and 57 (30.2%) females, and the control group consisted of 25 males (49.0%) and 26 females (51%). Patients’ CK-18 M30 levels were higher than values of the control group (308 [1–762] vs. 168 [67–287], P=0.001). Serum MMP-2 levels were found to be statistically higher in the patient group with respect to the controls (3.0 [1.1–6.8] vs. 2.0 [1.2–3.4], P=0.001). The highest serum CK-18 M30 and MMP-2 levels were measured in patients with cirrhosis. Serum apoptotic CK-18 M30 levels positively correlated with advanced age, fibrosis stage, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels (P= 0.001, 0.033, 0.001, and 0.001, respectively). Serum MMP-2 levels positively correlated with fibrosis stage, serum ALT, and AST levels (P= 0.001, 0.001, and 0.001, respectively).ConclusionsOur study indicated that CK-18 M30 and MMP-2 levels were higher in CHB patients compared to healthy controls and they were in association with significant hepatic fibrosis, especially cirrhosis.

Evaluation of the spontaneous reversibility of carbon tetrachloride-induced liver cirrhosis in rabbits

There is a general consensus that liver fibrosis in humans is potentially reversible, while scepticism prevails on the concept that cirrhosis can be truly reversed. The availability of suitable experimental models is fundamental for disease research. The experimental murine model of liver cirrhosis induced by carbon tetrachloride (CCl(4)) reproduces both the histological picture of the postnecrotic cirrhosis and its biochemical and clinical parameters. Normal hepatic structure is modified by formation of regeneration nodules. Fibrosis represents a morphological element of disease and an effect of hepatocyte necrosis. However, the relevance for research of this well-established model of liver cirrhosis is hampered by some spontaneous cirrhosis regression reported in mice and rats. It has been reported that CCl(4) also induces experimental liver cirrhosis in rabbits, but it is not known whether the process is reversible in this species. The aim of our study was to investigate this question. Male New Zealand White rabbits were treated intragastrically with CCl(4) or the vehicle only for 19 weeks and groups were sacrificed three and five months after treatment interruption. Cirrhotic and control livers were processed for routine light microscopy and for morphometric study of fibrosis by semiquantitative evaluation. The degree of fibrosis was based on the Knodell's scoring system.

Serum complement C4 in chronic hepatitis C: Correlation with histopathologic findings and disease activity

Hepatitis C virus leads to chronic liver disease, cirrhosis and hepatocellular cancer. Viral markers and other laboratory tests used in the diagnosis and follow-up of chronic hepatitis C do not correlate well with disease activity and liver histopathology. Therefore, alternative tests that indicate disease activity and relate with liver biopsy findings are needed. We aimed to investigate the relationship between serum complement levels and biopsy findings in patients with chronic hepatitis C. One hundred cases (70 patients, 30 healthy controls) were included in the study. Patients were divided into two groups: chronic hepatitis C patients with high transaminase levels were evaluated as the first group and patients with normal transaminase levels as the second group. Patients with a high transaminase level were biopsied and activity scores were evaluated against complement C3c and C4 levels. In addition, demographic data and laboratory tests were evaluated. Patients with chronic hepatitis C without proteinuria, acute phase response, cirrhosis, or coinfection with another hepatitis virus were included in the prospective study. Serum complement C3c (p<0.01) and C4 (p<0.01) levels were significantly lower in the first group than the second group. Serum complement C3c levels did not correlate with laboratory tests, hepatitis C virus-RNA levels, histological activity index, or fibrosis scores in patients with high transaminase levels, whereas complement C4 levels showed significant correlation with alanine aminotransferase (r: -0.368, p: 0.001) and histological activity index (r: -0.639, p: 0.001). We could not find any relation between serum complement C4 level and fibrosis. Serum complement C4 levels correlate with the histological activity index of the Knodell scoring system.

Lipid Profiles and Hepatitis C Viral Markers in HCV-Infected Thalassemic Patients

Background/AimsThe distribution of blood lipids, glucose and their determinants in thalassemic patients with chronic hepatitis C virus (HCV) infection has rarely been investigated. Thus, we aimed to investigate the relationship between both liver histologic findings and viral markers and serum lipids in thalassemic patients chronically infected with HCV.MethodsWe enrolled 280 polytransfused thalassemic patients with chronic hepatitis C. HCV viral load was determined using the Amplicor test. Genotyping was performed using genotype specific primers. Fasting serum lipid, glucose, ferritin and liver function enzyme concentrations were measured. A modified Knodell scoring system was used to stage liver fibrosis and to grade necroinflammatory activity. Perls' staining was used to assess hepatic siderosis.ResultsJust one subject had total cholesterol >200 mg/dL, and 7% had triglycerides >150 mg/dL. The mean high-density lipoprotein cholesterol (HDL-C) and glucose levels were 37 and 104 (97-111) mg/dL, respectively. Viral markers, liver histological findings and aminotransferase activity were not associated with serum lipid levels. Serum triglycerides, total cholesterol and ferritin were independent risk factors for impaired glucose tolerance or diabetes in these patients.ConclusionsThe majority of the patients had blood lipid levels (with the exception of HDL) within the defined normal range; viral and liver histological factors do not appear to play a significant role in changing the levels of serum lipids or glucose in these patients.

Amniotic Membrane Application Reduces Liver Fibrosis in a Bile Duct Ligation Rat Model

Biliary fibrosis and resultant cirrhosis are among the most common outcomes of chronic liver diseases. Currently, liver transplantation remains the only effective treatment. In seeking alternative therapeutic approaches, we focused on the potential use of the human amniotic membrane (AM). Indeed, AM has gained increasing importance for its antiscarring, anti-inflammatory, and wound-healing properties, as well as for the multipotent differentiation ability and immunomodulatory features of AM-derived cells. Intriguingly, we have recently demonstrated that placenta-derived cells reduce lung fibrosis in bleomycin-treated mice, and that AM patches reduce postischemic cardiac injury in rats. Hence, we have now investigated the effects of human AM on biliary fibrosis induced in rats through the bile duct ligation (BDL) procedure. A fragment of human AM was applied onto the liver surface after BDL and the effects on fibrosis establishment and progression were evaluated at different time points in comparison with fibrosis progression in control BDL rats. The degree of liver fibrosis was first assessed by the semiquantitative Knodell scoring system and, thereafter, by digital image morphometric analysis to quantify the area occupied by ductular reaction, activated myofibroblasts, and collagen deposition. We demonstrated a significant reduction in the severity of BDL-induced fibrosis in AM-treated rats. Indeed, while fibrosis progressed rapidly in control BDL rats, leading to cirrhosis within 6 weeks, AM-treated rats showed confined fibrosis at the portal/periportal area with no signs of cirrhosis, and a reduction in collagen deposition to about 50% of levels observed in control BDL rats. In addition, the AM was able to significantly slow the gradual progression of the ductular reaction and reduce, at all time points, the area occupied by activated myofibroblasts. These findings suggest that human AM, when applied as a patch onto the liver surface, might inhibit fibrosis progression in BDL-injured livers, and could protect against hepatic damage associated with fibrotic degeneration.

Prospective validation of a noninvasive index for predicting liver fibrosis in hepatitis C virus-infected liver transplant recipients

We previously developed a mathematical model, the Hospital Universitario La Fe (HULF) index, as an alternative to protocol liver biopsy (PLB) to estimate significant fibrosis (SF) in patients who underwent liver transplantation (LT) for liver damage caused by chronic HCV infection. In the present study, we sought to validate this noninvasive index. The commonly derived clinical and laboratory data for calculating the HULF index were prospectively collected over 2.7 years from patients undergoing LT and PLB. The sensitivity, specificity, positive and negative predictive values, and diagnostic capacity were evaluated with receiver operating characteristic curve analysis. Biopsy was performed 93 times in 86 LT patients. The prevalence of SF (F3-F4 on the Knodell scoring system) was 32%. The intraobserver and interobserver concordance was high (kappa = 0.94 and kappa = 0.75, respectively) in identifying SF in PLB. For low scores, the HULF index discarded an SF diagnosis with a sensitivity of 90% and a negative predictive value of 89%. The area under the receiver operating characteristic curve was 0.68. The precision of the HULF index did not improve with the incorporation of donor age and body mass index into the multivariate analysis. Applying the index would have prevented 24% of the biopsy procedures performed. In conclusion, the HULF index was prospectively validated with data commonly obtained in standard clinical practice. Because the index distinguishes a subgroup of HCV LT patients with a low probability of having SF, PLB would be avoided in those patients.

Effect of pegylated interferon beta-1a therapy on the liver fibrosis in chronic hepatitis C: a semi-quantitative analysis

To access the effect of pegylated interferon (PEGIFN) beta-1a on the reduction of liver fibrosis in chronic hepatitis C (HVC). Twenty-one patients with chronic HVC were divided into two groups randomly and treated with recombinant human PEGIHN-beta-1a (n = 13) or PEGIHN-beta-1a plus ribavirin (RBV) (n = 8) for 24 weeks, and then followed up for another 24 weeks. The clinical manifestations were observed and the clinical effects were evaluated. Biopsy was conducted before and after the treatment to analyze the histological activity index (HAI) and staging of fibrosis according the modified Knodell scoring system. Immunohistochemical analysis was used to examine the levels of alpha-smooth muscle actin (alpha-SMA), marker of activation of hepatic stellate cells (HSCs), and collagen type III in the HSCs. Sustained viral response (SVR) was achieved in 7 patients, and end-of-treatment virologic response (ETVR) was achieved in 9 patients. The HAI after treatment was 4.3 +/- 2.2, significantly lower than that before treatment (6.6 +/- 2.2, t = 4.77, P < 0.01). The fibrosis score after treatment was 1.1 +/- 1.1, significantly lower than that before treatment (1.7 +/- 1.2, t = 1.92, P < 0.05). The alpha-SMA score after treatment was 14 +/- 8, significantly lower than that before treatment (20 +/- 11, t = 2.15, P < 0.05). The integrated light density of collagen type III after treatment was 10 +/- 10, significantly lower than that before treatment (16 +/- 12, t = 1.83, P < 0.05). The improvement levels of fibrosis, alpha-SMA score, and integrated light density of collagen type III of the patients with SVR were all better than those of the patients without SVR; however, the differences were all not significant. The patients with combination therapy, female patients, and the patients with the HCV RNA < 2 x 10(6) copies/ml before treatment all showed higher levels in histology response. HAI, alpha-SMA level, and collagen type III values were all significantly correlated with the values of the semiquantitative indexes of fibrosis (all P < 0.01). Resisting hepatitis virus C and inhibiting and reversing the fibrotic progress, IFN-beta-1a therapy improves the liver histology of chronic HVC regardless of the viral response.

Effect of Angiotensin-converting Enzyme Inhibition on Experimental Hepatic Fibrogenesis

The renin-angiotensin system is suggested to be important in liver fibrogenesis. It induces hepatic stellate cell proliferation and up-regulates transforming growth factor beta-1 (TGF-beta1) expression. Matrix metalloproteinase-2 (MMP-2) is involved in extracellular matrix remodelling. Fibrosis, a consequence of most chronic liver diseases, may be the result of a disturbed balance between fibrogenesis and fibrolysis. The aim of this study was to investigate the effect of enalapril on liver fibrogenesis induced in rats by bile-duct ligation. Forty-seven rats were divided into two groups: bile-duct ligated (BDL) (n = 24) and BDL + enalapril (n = 23). Fibrosis was evaluated by the Knodell scoring system, and TGF-beta1 and MMP-2 were assessed with immunohistochemistry at the second, fourth and sixth weeks after bile-duct ligation. In the BDL group, TGF-beta1 increased by the second week and this increase continued through weeks 4 and 6. In the BDL + enalapril group, TGF-beta1 was significantly lower than the other group (P < 0.05). MMP-2 progressively decreased after week 2 in the BDL group. In the BDL + enalapril group, MMP-2 was significantly higher than the BDL group at the fourth and sixth weeks. These results suggest that enalapril reduces the liver tissue TGF-beta1 and has an ameliorating effect on the fibrosis markers TGF-beta1 and MMP-2.

What is the reason of elevated alanine aminotransferase level in HBeAg negative patients with low viremia: NAFLD or chronic hepatitis?

Background and study aims: Increased alanine aminotransferase (ALT) levels with negative hepatitis B virus (HBV) DNA by hybridization is a common problem in Turkey where is a mild endemic region. We aimed to evaluate the causes of elevated ALT levels in patients who are negative for hepatitis B e antigen (HBeAg) and HBV DNA (by hybridization) for at least 6 months.Patients-methods: Forty-nine patients were enrolled in this study. Histological changes [histological activity index (HAI), and the extent of fibrosis] were assessed according to the Knodell scoring system and steatosis were graded by Brunt’s classification for NAFLD in all patients.Results: A mean age of the patients was 34.9 ± 12.1 years (16-70). 43 (87.8%) of them were male. Mean ALT level was 95 ± 39.7 IU/L (50258). Hyperglycemia (>100 mg/dL) and hyperlipidemia were found in 12 and 24 patients, respectively. Hepatic steatosis (7 patients grade 1; 5 patients grade 2; and 7 patients grade 3), ground-glass hepatocyte, chronic hepatitis, and Wilson disease were found in liver biopsy in 38.8%, 32.6%, 26.6%, 2%, respectively. Mean HAI was 6.5 ± 3.6 (4-12) in chronic hepatitis. Seven patients (53.9%) were in stage 1 and 2 while 6 patients (46.1%) were in stage 3 and 4.Conclusions: Nonalcoholic fatty liver disease is the most common cause of elevated ALT levels in HBeAg negative/HBV DNA negative patients. Chronic hepatitis B was found in 26.6% of these patients.

Apoptosis in chronic viral hepatitis parallels histological activity: An immunohistochemical investigation using anti‐activated caspase‐3 and M30 Cytodeath antibody

Apoptosis is implicated as a major pathogenic mechanism in chronic hepatitis B and C. Previous studies of the relationship between apoptotic rates and histological necroinflammatory activity have produced conflicting results. Hepatocyte apoptosis was assessed in liver tissue from 32 cases of chronic viral hepatitis, seven cases of hepatocellular carcinoma (HCC) and six cases of steatohepatitis as non-viral disease controls and eight cases of control liver. Apoptotic rates were measured using H&E morphological assessment and immunohistochemical staining with antibodies to activated caspase-3 and M30. Histological necroinflammatory activity of viral hepatitis cases was scored using the Knodell scoring system, and the cases were divided according to their score into group 1 (mean 2.43 +/- 0.48) and group 2 (mean 7.80 +/- 0.49). Apoptotic indices were significantly higher in group 2 than group 1 using H&E (11.53 +/- 2.70 vs. 0 +/- 0, P=0.015) and activated caspase-3 (22.01 +/- 5.27 vs. 1.79 +/- 1.79, P=0.03) methods but were not significantly higher with M30 (3.80 +/- 1.74 vs. 0 +/- 0, P=0.207). Apoptotic scores using an antibody to activated caspase-3 are significantly higher in cases of chronic viral hepatitis with greater histological necroinflammatory scores, supporting a central role for apoptosis in disease pathogenesis. This method offers an alternative to routine histological assessment for measuring disease activity.

Histological spectrum of chronic hepatitis in precore mutants and wild-type hepatitis B virus infection.

This study compares the histology of liver biopsies in precore mutant (PCM) and wild-type hepatitis B virus (HBV) infection. Thirty cases of PCM and 60 cases of wild-type HBV infection were included. Patients were diagnosed on the basis of serological profile and HBV DNA assessment. Liver biopsies in each case were assessed for histological activity and stage of fibrosis using a modification of Knodell's scoring system. There was no statistically significant (P=0.14) difference in histological activity in the two groups. The difference in stage of fibrosis in the two groups was statistically significant (P=0.001). Quantitative estimation of viral load did not show any correlation with liver histology. PCM HBV showed greater stage of fibrosis compared with wild-type infection, and this may be related to disease progression and lack of response to therapy.

Assessment of histological features of changes in lipid metabolism in chronic hepatitis C

Introduction: Several data suggest interactions between HCV and lipid metabolism. Nevertheless steatosis is a frequent histological lesion in CH-C, this phenomenon was not analyze as an integral part of the grade in the modified Knodell scoring system.

Expression of Fas antigen in liver tissue of patients with chronic hepatitis B and C.

Cells infected with the hepatitis B or C virus can be eliminated by an immune response mediated by cytotoxic T cells. Recently, the Fas Ligand has been detected on the surface of cytotoxic T cells, and is thought to induce cells to apoptosis by adhering to the Fas antigen. To evaluate the role of the Fas antigen and apoptosis in chronic hepatitis B or C, we used immunohistochemistry to study Fas antigen expression in liver samples obtained from 30 patients infected with chronic hepatitis B and 32 patients with chronic hepatitis C. In samples from patients with chronic hepatitis C and B, Fas antigen was mainly expressed in the cytoplasm (partly at the membrane) of hepatocytes, and these positive cells were detected especially at the periportal region near 'piecemeal necrosis'. According to Knodell's scoring system for the histological activity index, the scores of periportal inflammation and necrosis were higher in cases that were positive for Fas antigen than in cases that were negative for the antigen (n = 62, P < 0.001) and there was a positive correlation between these scores and the degree of Fas antigen expression (r = 0.621). There was also a positive correlation between the scores of intralobular inflammation and necrosis and the degree of Fas antigen expression (n = 62, r = 0.522, P = 0.001). Fas antigen was not detected in patients without hepatitis infection. These findings suggest that apoptosis of hepatocytes is induced via the Fas antigen and contributes to the elimination of infected cells.

Effects of PPARg agonist pioglitazone on rat hepatic fibrosis.

To investigate effects of pioglitazone on rat hepatic fibrosis and to explore its mechanism. Rat hepatic fibrosis was induced by carbon tetrachloride (CCl(4)). Forty Sprague-Dawley rats were divided randomly into 4 groups: control, model, and two treatment (PI, PII) groups. Except for rats in control group, all rats were given subcutaneous injection of 400 mL/L CCl(4), twice a wk for 8 wk. Rats in PI and PII groups were also treated with pioglitazone of 3 mg/kg, daily via gastrogavage beginning on the 1(st) day and at the end of the 2(nd) week, administration of CCl(4) respectively. Liver functions (ALT, AST), serum fibrotic markers (HA, LN, PCIII) and hepatic hydroxyproline (HP) concentration were determined respectively. Histochemical staining of formalin-fixed liver sections with HE, Masson-Trichrome, and immunohistochemical staining for alpha-smooth muscle actin (alpha-SMA) were performed. Modified Knodell and Chevallier semi-quantitative scoring system (SSS) was used to evaluate necroinflammatory activity and fibrosis degree. Compared with model group, pioglitazone significantly reduced the serum levels of ALT, AST, HA, LN and PCIII (P<0.05 or <0.01). The HP concentrations in PI (210.90+/-24.07 microg/g), and PII (257.36+/-30.55 microg/g) groups were also lower than those in model group (317.80+/-36.44 microg/g) (P<0.01). Histologic examination showed that PI and PII groups had milder hepatocellular degeneration, necrosis and infiltration of inflammatory cells, and thinner or less fibrotic septa than did model group. The scores for necroinflammation in PI (2.80+/-1.03), and PII (3.00+/-1.05) groups were significantly reduced as compared with model group (4.88+/-2.30) (P<0.05 or <0.01); the fibrosis scores in PI (3.40+/-1.65), and PII (4.60+/-1.35) groups were also markedly lower than those in model group (7.00+/-3.21) (P<0.05 or <0.01). Immunohistochemical staining showed that expression of alpha-SMA in PI and PII groups was ameliorated dramatically compared with model group. PPARgamma agonist pioglitazone greatly retards the progression of rat hepatic fibrosis induced by CCl(4) through inhibition of HSC activation and amelioration of hepatocyte necroinflammation in rats.

Effect of angiotensin II type 1 receptor blockade on experimental hepatic fibrogenesis

Background/Aims : The aim of this study was to investigate whether in the liver, as in other tissues, there is evidence that angiotensin II, acting via the angiotensin II type 1 receptor (AT1-R), plays a role in fibrogenesis. Methods : Sprague–Dawley rats were divided into three groups; sham, bile duct ligated (BDL) and BDL+AT1-R antagonist, irbesartan. Real time RT-PCR was utilised to assess gene expression of the AT1 receptor, TGF-β1 and α1 (I) collagen in the liver. TGF-β1 and α1 (I) collagen mRNA expression and localisation were also assessed by in situ hybridisation. TGF-β1 activity was assessed by using the TGF-β inducible gene product βig-h3. Fibrosis was assessed by the Knodell scoring system, tissue hydroxyproline content and picro-sirius red staining. Results : Real time RT-PCR revealed that there was a 6-fold up-regulation in AT1 receptor expression in BDL animals compared with shams. This was associated with marked increases in TGF-β1, βig-h3 and α1 (I) collagen gene expression which were attenuated by AT1-RA treatment. However, AT1-RA therapy produced no significant change in liver histology or hydroxyproline content. Conclusions : These results suggest that in the liver angiotensin II may play an important role in the fibrogenic response to injury. However, whether treatment with an AT1-RA will be of therapeutic benefit remains to be determined.

Influence of Liver Fibrosis on Hepatic Artery Doppler Resistance Index in Chronic Hepatitis of Viral Origin

Hepatic arterial Doppler sonography is increasingly being used in liver diagnostics. The determinants of the elevation of hepatic artery impedance indexes in chronic liver disease, however, have still not been fully clarified. The aim of the present study was to investigate the relationship between histological alterations and liver circulation in chronic hepatitis. Hepatic artery resistance index and portal flow velocity were measured using Doppler sonography in 47 patients with chronic hepatitis of viral origin diagnosed at histopathology. The patients were divided into two groups, those with mild and those with severe alterations, in accordance with the various histological parameters of the Knodell scoring system. Hepatic artery resistance index and age were higher in patients with more severe liver fibrosis (respectively 0.638 +/- 0.084 and 39.0 +/- 10.9 (years) in mild fibrosis versus 0.687 +/- 0.060 and 49.4 +/- 14.4 (years) in severe fibrosis; P < 0.05 for both), whereas no difference between the two groups was found for the other histological features (degeneration, inflammation and necrosis), nor for portal flow velocity. The increase in hepatic artery resistance index appears to be influenced by the extent of fibrous tissue deposition in the liver, determined by chronic inflammation and repair and, secondly, by ageing.

High levels of hepatitis C virus RNA in native livers correlate with the development of cholestatic hepatitis in liver allografts and a poor outcome.

A subset of hepatitis C virus (HCV)-positive liver transplant recipients develop cholestatic hepatitis (CH). We investigated the role of pretransplantation disease activity (estimated by Knodell score and HCV RNA quantitation) in the native liver explant on the development of CH and graft and patient outcome. Eight patients with CH were identified among HCV-positive liver transplants and were compared with 20 consecutive patients with recurrent HCV hepatitis of noncholestatic type in liver transplants. We evaluated all 28 explanted native livers histologically using the Knodell scoring system. HCV viral load was measured in the native explant and 5 allograft explants from the CH group using Amplicor HCV RNA Monitor test. Six of 8 patients with CH had HCV RNA levels of 5,000 copies/microg of DNA or greater in the native liver explant, whereas only 1 of the control group had viral loads greater than this level. Greater HCV RNA levels correlated with worse graft and patient survival (P <.001). The 3-year survival rate in the CH group was 18% compared with 77% in the control group (P <.001). There was no difference in the primary immunosuppressive regimens used in the 2 groups. We conclude that (1) CH has a uniformly poor prognosis, (2) type of immunosuppressive therapy appears to have little influence on the development of CH, (3) high pretransplantation HCV RNA levels in the native explant may predict the development of CH, and (4) patients with high HCV RNA levels in the explanted native liver may be appropriate candidates for antiviral therapy to prevent the development of CH.