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المستخلص Hepatitis C virus (HCV) infection is a major health problem worldwide that causes chronic hepatitis, liver fibrosis, cirrhosis, or even hepatocellular carcinoma (HCC). Because the liver is regarded as the body’s metabolic powerhouse, it is densely packed with mitochondria that support its high energy demands. Hepatitis viruses can modify mitochondrial morphology and function by regulating the number, quality, and dynamics of mitochondria. chronic HCV infection has been linked to hepatic mitochondrial injury, which includes swelling, loss of mitochondrial cristae, decreased mitochondrial number, calcium-mediated mitochondrial depolarization and malfunction, and the production of reactive oxygen species (ROS). The severity of HCV infection is represented by the extent of mitochondrial injury as it leads to overproduction of ROS which is now recognized as a major pro-carcinogenic cofactor in chronic HCV infection. The blood of those with persistent HCV infection and those with HCC had a low copy number of mitochondrial DNA (mtDNA). Because a decreased level of serum mtDNA content is linked to an increased HCC risk, serum mtDNA could be used as a noninvasive predictor of HCC risk. In the past two decades, patients with chronic hepatitis C (CHC) have been widely treated with interferon (IFN)-based therapy, however, this led to only low rates of sustained virologic response (SVR). Treatment of HCV has evolved with the development of direct-acting antivirals (DAAs) which allowed for simplified regimens with increased tolerability and wider therapeutic window and has radically improved the SVR rates even for patients with cirrhosis. IFN treatment in HCV patients significantly reduced the rate of mtDNA mutations in hepatocytes and increased mtDNA content in peripheral leukocytes. Furthermore, it’s still unknown if SVR after DAAs treatment results in reversal of mitochondrial dysfunction or not. Therefore, the present work was designed to identify the impact of DAAs on mtDNA copy number in patients with HCV-related chronic liver disease. Fifty patients with chronic HCV infection and 20 age- and sex-matched healthy subjects with no liver disease were enrolled in the present study. All patients tested positive for HCV antibody and had detectable HCV RNA in their blood. The diagnosis of HCV-related cirrhosis was based on the clinical evaluation, laboratory workup, and radiological features. Patients were excluded from the study if they had hepatitis B virus (HBV) or human immunodeficiency virus (HIV) co-infection, other causes of chronic liver disease, decompensated chronic liver disease, advanced cardiopulmonary disease, malignant disease or pregnancy. Eligible patients were offered DAAs for 12 weeks duration according to the clinical guidelines for management of hepatitis C. The study followed the provisions of the Declaration of Helsinki as well as the Good Clinical Practice recommendations. All of the participants in the study gave their informed consent. All patients with chronic HCV infection were evaluated clinically as regards the symptoms and signs of chronic liver disease (chronic fatigue, right hypochondrial pain) if present, and liver and spleen size. Abdominal ultrasonography was performed for assessment of liver size and echopattern, and the presence of cirrhosis, ascites and splenomegaly. Laboratory investigations were done in all patients and healthy subjects including complete blood picture, serum creatinine, serum glucose, liver test profile [serum alanine and aspartate aminotransferases (ALT and AST respectively), serum bilirubin, serum albumin and international normalized ratio (INR)]. Serum HCV RNA and serum mtDNA content were measured using quantitative real-time polymerase chain reaction (PCR) before the start of treatment as a baseline, and 12 weeks after the end of treatment. All patients were treated with DAAs-based treatment regimens including Sofosbuvir + Daclatasvir ± Ribavirin for 12 weeks duration. Treatment response was monitored by measurement of HCV RNA and serum mtDNA copy number at 12 weeks after the end of treatment. When HCV RNA was less than the lower limit of detection at week 12 after treatment, it was considered as sustained virological response (SVR12). All our patients achieved SVR12. Real-time quantitative PCR was used to assess the number of copies of the mitochondrial ND1 gene, and the human globulin gene was included as a reference gene. Statistical analysis of the obtained data showed the following results: • Serum HCV-RNA level ranged between 1.55– 6140.0 x103 IU/ml in patients (mean ± SD: 969.3 ± 1374.4 IU/ml). • Serum levels of AST and ALT ranged between 20–127 U/L and 15–139 U/L respectively in patients and between 12–26 U/L and 5–27 U/L respectively in healthy subjects. The mean serum AST and ALT levels in patients were significantly greater than in healthy volunteers (P < 0.001 each). • Serum albumin concentration ranged between 2.8–4.8 g/dl in patients while it ranged between 3.9–4.5 g/dl in healthy subjects. The mean albumin concentration showed a significant decrease in patients compared with that in healthy subjects (3.72 ± 0.32 g/dl vs. 4.23 ± 0.19 g/dl, P < 0.001). • Serum bilirubin level ranged between 0.1–3.5 mg/dl in patients while it ranged between 0.3–0.9 mg/dl in healthy subjects. There was no statistically significant discrepancy between patients and healthy subjects (0.53 ± 0.47 mg/dl vs. 0.55 ± 0.19 mg/dl, P = 0.200). • INR ranged between 0.9–1.51 in patients while it ranged between 0.88–1.12 in healthy subjects. Between patients and healthy volunteers, there was no significant disparity. (1.03 ± 0.10 vs. 1.0 ± 0.07, P = 0.106). • Serum alpha fetoprotein (AFP) level ranged between 0.1– 39.5 ng/ml in patients, while it was ranged between 1–3.4 in healthy subjects. There was no statistically significant disparity between patients and healthy persons (6.07 ± 7.03 ng/ml vs. 2.37 ± 0.7 ng/ml, P = 0.096) • FIB-4 ranged between 0.27 –3.58 in patients and the mean was 1.55 ± 0.73. FIB-4 of ≥ 1.45 was found in 27 patients (54%) and <1.45 in 23 patients (46%). Meanwhile, APRI ranged between 0.14-1.81 in patients and the mean was 0.56 ± 0.30. • The liver echopattern was normal in 40 patients (80%), and coarse in 10 patients (20%). • As regard the treatment regimen, 46 of the patients (92%) received Sofosbuvir + Dacltasvir for 12 weeks, while 4 of the patients (8%) received Sofosbuvir + Daclatasvir + Ribavirin for 12 weeks. • Serum mtDNA copy number ranged between 9.38–102.5 in the patients before treatment of HCV infection and changed to 12.99–104.0 after treatment, while it ranged between 24.02–116.7 in healthy subjects. The mean serum mtDNA copy number was significantly lower in HCV-infected patients before treatment compared with healthy subjects (25.30 ± 15.28 vs. 45.05 ± 22.03, P < 0.001). Also, there was significant increase in the mean serum mtDNA copy number in patients after treatment compared to the mean levels before treatment (38.07 ± 21.46 vs. 25.30 ± 15.28, P < 0.001) while there was no statistically significant difference between patients after treatment and healthy subjects (P = 0.059). • The statistical correlations between serum mtDNA copy number (2-ΔCt) in patients with chronic HCV infection on one hand and other studied parameters on the other hand showed the following results: Serum mtDNA copy number (2-ΔCt) was inversely correlated with serum bilirubin level (r = -0.348, P 0.013). There were no statistically significant correlations between mtDNA copy number (2-ΔCt) on one hand and age, hemoglobin concentration, platelet count, blood glucose, serum creatinine, ALT, AST, serum albumin, INR, HCV RNA, FIB-4, APRI and AFP on the other hand (P > 0.05). |