27- Monto A, Alonzo J, Watson JJ, Grunfeld C, Wright TL. Steatosis in chronic hepatitis ... 34- Potter JJ, Womack L, Mezey E, Anania FA. Transdifferentiation of rat.
AAMJ, Vol. 10, N. 3, Sep, 2012, Suppl. ــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ
ESTIMATION OF SERUM LEPTIN LEVELS IN PATIENTS WITH CHRONIC HEPATITIS C Khaled A. Eid1, and Mona M. Abdel Maguid 2 Hepatology and Gastroenterology Department, .Faculty of Medicine – AlAzhar University – Assiut1 and Clinical Pathology Department, Qena Faculty of Medicine – South Valley University2. ــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ ABSTRACT Objectives: The adipose tissue can play active proinflammatory and profibrogenic role in pathogenesis of chronic hepatitis C (CHC). Leptin seems to play important, although not fully elucidated, role in pathogenesis of liver damage. Aim of the work: to investigate the relationship between serum leptin concentrations and the degree of hepatic fibrosis in patients infected with hepatitis C virus and to assess whether leptin level is affected by the combined antiviral treatment treatment when estimated 12 weeks after the start of treatment. Patients and methods: 40 patients with CHC together with 10 healthy control subjects were enrolled in the study. Leptin level was estimated in these subjects at baseline and after 12 weeks from the start of combined antiviral therapy. Patients had liver biopsy taken at baseline to determine the fibrosis and inflammation grades according to the METAVIR study, 1994 [41]. Results: Leptin concentration at baseline showed no significant difference between patients and control group (P> 0.05). Leptin level was much higher in female patients (36.1 ± 23.6) than in males (10.4 ± 9.9) (P< 0.05) at baseline group but not in the treated group. A high significant positive correlation was found between serum leptin level, inflammation grades and fibrosis stages (r= 0.810, p < 0.001 & r= 0.791, p < 0.001 respectively). Serum leptin concentrations in patients treated by 157
Khaled A. Eid and Mona M. Abdel Maguid
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combined antiviral therapy for 12 weeks is decreased more than baseline group with a mean ± SD of (8.8 ± 11.1 & 14.9 ± 16.3 respectively) although not reaching a statistically significant levels (P=0.054). Conclusion: Leptin has a profibrogenic and/or proinflammatory role as higher leptin levels were correlated with fibrosis stages and inflammation grades in CHC patients. Interferon-alpha antiviral therapy can decrease and suppress leptin levels in CHC patients. INTRODUCTION In the last two decades a great progress was made both in our understanding of viral pathogenesis and clinical course of chronic hepatitis C virus infection (HCV) as well as in antiviral therapies of this disease [39]. HCV infection frequently leads to chronic hepatitis, which may progress to cirrhosis and even to hepatocellular carcinoma (HCC) [29]. Some factors associated with rapid progression of liver injury and fibrosis were identified and may be divided into non-modifiable (age, gender, virus genotype) and potentially modifiable ones (metabolic factors, like steatosis and insulin resistance) [6]. Leptin, the adipocyte-derived protein product of the ob gene, is involved in appetite regulation and obesity through central effects at the hypothalamus [17]. Leptin is related to amount of body fat [9].
The
importance of leptin in the regulation of energy balance, body composition, and food intake has been demonstrated in both animal and human studies [12,18,47,]. Leptin has been implicated liver fibrogenesis. Serum leptin levels have been found higher in patients with chronic hepatitis C (CHC) and particular in those with more severe fibrosis or cirrhosis [5]; however the results are conflicting [40,4,8,15,16,30,40,44]. Non-alcoholic fatty liver disease (NAFLD) represents a spectrum of liver diseases characterized mainly by macrovesicular steatosis that occurs in the 158
AAMJ, Vol. 10, N. 3, Sep, 2012, Suppl. ــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ absence of alcoholic consumption. The hepatic histology aries from isolated hepatic steatosis alone “first hit” to fatty liver accompanied by hepatocellular damage plus inflammation known as steatohepatitis “second hit” which is followed by the development of fibrosis [27]. In this study, serum leptin levels in CHC patients at baseline were investigated and their relationships with histopathological fibrosis and inflammation grades were assessed. Leptin level was estimated also 12 weeks after the start of antiviral treatment in the same patients to assess whether it is decreased by the combined antiviral therapy or not to elucidate the role of leptin in CHC patients and its potential clinical significance.
PATIENTS AND METHODS Forty patients suffering from CHC infection who were subjected for combined antiviral treatment were enrolled in this study; they were evaluated to Qena center for CHC treatment with interferon and ribavirin. Also, 10 normal healthy control subjects were included in the study matched for age and sex. The patients were divided into 2 groups: patients at baseline (having CHC before the start of treatment) and the same patients 12 weeks after the start of combined antiviral treatment. Exclusion criteria included patients with other causes of chronic liver disease, decompensated liver disease, and history of heart failure, diabetes, thyroid diseases, abnormal renal function, cancer, previous treatment with antiviral agents, and the use of drugs known to induce liver steatosis within the last 6 months. The study protocol had been approved by the Ethics Committee and patients gave oral consent to participate in the present study. All patients and controls were subjected to clinical assessment and abdominal ultrasonography.
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Khaled A. Eid and Mona M. Abdel Maguid
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Baseline characteristics collected at the time of liver biopsy included sex, age, height, and weight. Body mass index is calculated. Serological tests: Anti-HCV, HBsAg & Anti-HIV Abs were determined by ELIZA. Complete blood count, prothrombin time and INR & liver enzymes (ALT, AST) and total bilirubin were estimated. Liver biopsy was done for all patients. Liver specimens were formalin-fixed and paraffin-embedded for histological evaluation. Stained sections were evaluated according to a scoring system that includes the semi-quantitative assessment of liver disease grading and staging (METAVIR study, 1994) [41]. Determination of Leptin level in serum: in patients it was done before and after 12 weeks from the completion of treatment. Leptin estimation was done by ELISA kit from Labor Diagnostika Nord GmbH & Co. KG (LDN). Five ml of venous blood were drawn from every individual after an overnight fast in glass collecting tubes with no anticoagulants added. Blood was centrifuged for 4 minutes and serum was aliquoted, separated and stored at -80oC till serum Leptin was estimated. Figures > 11.1 ng/ml in females and > 5.6 ng/ml in males were considered high. Statistical Methods: Analysis of data was done by IBM computer using SPSS (statistical program for social science version 12). All results were expressed as range (mean ± SD). RESULTS The study included 40 patients (33males and 7 females). BMI was (25.9 ± 3.3) kg/m2 (table 1). Serum leptin concentrations in all patients was increased (14.9±16.3) with no significant difference between patients and control group (P> 0.05) and is decreased in the second group after 12 weeks of treatment more than baseline group (8.8 ± 11.1 vs 14.9 ± 16.3) although not reaching a 160
AAMJ, Vol. 10, N. 3, Sep, 2012, Suppl. ــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ statistically significant levels (P=0.054) (table 2). Leptin level was much higher in female patients at baseline (36.1 ± 23.6) than in males (10.4 ± 9.9) with a statistical significant difference (P< 0.05) but not in the second group (table 3). Serum leptin level in the baseline group was positively correlated with BMI (r= 0.719, p < 0.001) (figure 1) and is a negatively correlated with AST & ALT and T. bilirubin (r- value = 0.029, 0.097 & 0.162 respectively and P-value = 0.839, 0.501 & 0.261 respectively) (table 4) whereas, in the after 12 ws group it was positively correlated with AST & ALT and T. bilirubin (r- value = 0.473, 0.398 & 0.308 respectively and P-value = 0.001, 0.001 & 0.030 respectively) (table 4). Liver histopathological examination revealed: 5 (12.5%) patients with no activity (A0), 19 (47.5%) with mild activity (A1), 15 (37.5%) with moderate activity (A2) and only 1 (2.5%) with severe activity (A3). Twinty (50%) of patients without fibrosis (F0), 16 (40%) with mild fibrosis (F1) and 3 (7.5%) with moderate fibrosis (F2) and only 1 (2.5%) with advanced fibrosis (F3) (table 5). There was a significant correlation between serum leptin level and histopathological inflammation grades & fibrosis stages (r= 0.810, P
