Circulating levels of cytochrome C, gamma-glutamyl transferase, triglycerides and unconjugated bilirubin in overweight/obese patients with non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease, characterized by hepatocyte apoptosis, is distinct in fatty liver and non-alcoholic steatohepatitis, the more severe form. Apoptotic cell death is caspase-dependent and associated with mitochondrial membrane depolarization and cytochrome c release. Adhering to the hypothesis that the exposure of hepatocytes to free fatty acids, resulting in increased ROS production and mitochondrial damage, is balanced by the presence of antioxidant substances, circulating levels of gamma-glutamyl transferase, cytochrome c, triglycerides and unconjugated bilirubin were explored in patients suffering from non-alcoholic fatty liver disease with different severity. One hundred and eighty-six consecutive patients who presented recent ultrasound feature of bright liver without any liver disease of known origin were enrolled, eighty-nine of whom underwent liver biopsy. Forty-five subjects were allocated on the basis of histology in fatty liver group while 44 patients formed the group with non-alcoholic steatohepatitis. A cohort of 27 young, lean, apparently healthy individuals was selected as control group. The levels of gamma-glutamyl transferase were normal or slightly increased, while unconjugated bilirubin concentrations were elevated in all the spectra of non-alcoholic fatty liver disease. Comparing the present results with relevant findings from other studies dealing with diseases characterized by apoptosis, we did not find high circulating levels of cytochrome c in non-alcoholic fatty liver disease. What is more, our patients, categorized as suffering from simple fatty liver or from the more severe non-alcoholic steatohepatitis, had similar levels of cytochrome c and gamma-glutamyl transferase, p=0.19 and 0.11. Serum triglycerides were higher in patients with non-alcoholic fatty liver disease than in the healthy group, p=0.001. These findings likely reflect a balance between oxidative stress and anti-oxidant response rather than a lack of reliability of cytochrome c as a reliable biomarker of mitochondrial damage.

Can serum TGF-beta 1 be used to evaluate the response to antiviral therapy of haemophilic patients with HCV-related chronic hepatitis?

Congenital coagulation disorders limit the use of liver biopsy, especially when repeated assessment is needed. TGF-beta 1 plays a pivotal role in inducing fibrosis and has been proposed as its surrogate marker. Aiming at validating the clinical utility of this cytokine, fifteen haemophilic patients suffering from HCV-related chronic hepatitis were treated with Peg-IFN alpha2beta plus Ribavirin. Serum TGFbeta 1, viral load and liver enzymes were analyzed at baseline and at six, twelve, and eighteen months. As expected, patients initially showed significantly higher TGF-beta 1 levels than age-matched controls (43.8 ng/mL, 28.7-46.4 vs. 26.9 ng/mL, 23.0-34.0, median and 95% CI; p=0.004). The end of therapy response rate was 67%. The main finding was a significant drop in TGF-beta 1 at six months compared to baseline values; this drop de facto predicted the levels reached in the following six months, which were fixed at lower concentrations (37.0 ng/mL, 21.9-43.8 and 27.0 ng/mL, 24.1-44.0 respectively; p<0.009), independently of treatment outcome (three patients were breakthrough, twelve were sustained virological responders (SVRs). During the treatment period none had clinical or biochemical signs of inflammation in other areas. Treatment was followed by a six-month follow-up, at the end of which TGF-beta 1 was increased compared to the previous values, reaching the initial levels in ten SVRs (45 ng/mL, 24.5-52.9). Interestingly, at a longer follow-up, two out of ten SVRs, who displayed the highest values of TGF-beta 1, relapsed. Serum TGF-beta 1 could be used to assess therapeutic outcome and short-term prognosis of HCV-related chronic hepatitis.

Serum concentrations of the tissue polypeptide specific antigen in patients suffering from non-alcoholic steatohepatitis.

BACKGROUND: Liver histology is the gold standard for diagnosis of non-alcoholic fatty liver disease. Ethical considerations and patient choice often preclude performing a liver biopsy, especially considering the rare but potential risk. Searching for a good serological marker substitute for the invasive procedure was the aim of our study. Keratins, mainly 8 and 18, play not only a mere structural role providing mechanical stability to hepatocytes, but also represent a target via toxic stress ultimately inducing apoptosis/necrosis. Tissue polypeptide-specific antigen (TPS), a serological mirror of keratin 18, is widely used as a marker for various cancers. This antigen was assessed in three different groups who were overweight or obese. MATERIALS AND METHODS: In this cross-sectional case-control study, 48 cancer-free patients with non-alcoholic steatohepatitis (NASH, Group 1), 48 patients with pure fatty liver (FL, Group 2), and 47 volunteers (Group 3) were studied. All of them were referred to our metabolic unit for routine evaluation. RESULTS: The median (range) TPS levels were 123 (56-286) ng mL(-1) in NASH patients. FL patients and volunteers had significantly lower TPS levels, 76 (38-98) ng mL(-1) and 64 (28-87) ng mL(-1), respectively (P = 0.0001). A value of 88 ng mL(-1) in patients with underlying bright liver was associated with a high probability of NASH (sensitivity and specificity = 92% and 96%, respectively). One patient (2.1%) with FL had a TPS value > 88 ng mL(-1), but in the same group, 29 FL patients (60.4%) had an alanine aminotransferase value > 40 U L(-1). Based on a recent classification of liver fibrosis, the median (range) TPS values were significantly different among the stages: F1 (n = 23) = 100 (76-264) ng mL(-1); F2 (n = 21) = 134 (56-276) ng mL(-1); and F3 (n = 4) = 199.5 (123-286) ng mL(-1), respectively (P = 0.014). CONCLUSIONS: Our study shows that TPS is a better marker than alanine aminotransferase activity, ultrasonography or the combination of both parameters in differentiating NASH from FL.

Hepatic and peripheral T-lymphocyte patterns in patients with chronic hepatitis C infection: what correlation with histological activity?

The immunopathological processes involved in hepatic damage during chronic hepatitis C infection are not fully understood. Several works suggest the role of T helper 1 (Th1) immune response in both injury and fibrinogenesis. in this study, we have analyzed peripheral and intrahepatic T-lymphocyte subsets in liver biopsy specimens of 13 patients with definite chronic hepatitis C (CHC) to explore the possible direct role of these patterns in the evolution of necrotic inflammation and fibrogenesis scored according to the Knodell histological activity index. in particular, we have studied CD4+/CD7+ T-lymphocytes, as phenotypic marker of Th1 immune response, CD4+/CD7- as Th2 marker, and CD8+/CD38 as activated CD8+ lymphocytes. on statistical analyses we found a significant negative correlation in liver CD8+/CD38+ T-cells grading (r= -0.607; p<0.05) and staging index (r= -0.650; p<0.05) and between CD4+/CD7+ and grading (r= -0.626; p<0.05) index. in addition, we found a positive strong correlation between CD38+/CD8+ and CD4+/CD7+ T cells (r= 0.783; p<0.05) in liver tissue and between peripheral and liver resident CD8+/CD38+ (r= 683; p<0.05). moreover, the hepatic CD4+/CD7+ T-cells showed a positive correlation with peripheral CD 8+/CD38+ T-cells (r= 0.676; p<0.05). A strong positive correlation was also observed between grading and staging index (r= 0.921; p<0.01). we found no statistical correlation between the above variables and CD4+/CD7- T cells. our data could suggest that a preferential hepatic CD4+/CD7+ OR CD8+/CD38+ T cell subset was not directly associated with hepatic damage but, on the contrary, it could have been able to block liver injury. Concerning the peripheral subsets, the only CD8+/CD 38+ T-cells result reflect the CTL activity in the liver tissue. further studies are required to better understand the possible correlation between peripheral and liver resident T-helper, subset and other hepatic resident immunocompetent cells.

CD8+/CD38+: immune activity and clinical significance in HCV patients with and without interferon therapy.

At the light of the importance of cytotoxic T lymphocyte (CTL) response during chronic hepatitis C, we carried out a study in order to evaluate the CD8+/CD38+T-cells, immunophenotypic marker of CD8+ activated cells in a selected cohort of 22 patients for four months. The patients were subdivided in two groups: A (with IFN therapy), B (without IFN therapy). The results show that in IFN-treated subjects there is a significant reduction of ALT (sign test, z = .424;p < or = .05) and that the CD8+/CD38+ present a positive correlation with HCV-RNA (r = .894; p < .05). We hypothesize that during IFN therapy the CD8+/CD38+ activity is able to oppose HCV, probably by increasing MHC I expression on the infected cells due to the IFN modulatory action, that could strengthen the immune response of CD8+ activated T-lymphocytes. These events confer the capacity to specifically respond to any viral replication and probably take part in the reduction of ALT levels by decreasing the chronic inflammation present during a defective immune response. These data show think CD8+/CD38+ marker could be a good parameter to evaluate both the viral activity and immunological status in HCV+ patients undergoing IFN treatment.