الفهرس 
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Abstract
The current study was carried out at Microbiology and Immunology department, Faculty of Medicine, Minia University, in the period from February 2017 to December 2019. It was conducted on 200 subjects divided into 2 groups. group І (HCV patients group), group ІI (control group). group I were 150 chronic HCV patients with positive anti-HCV antibodies by ELISA and HCV RNA by reverse transcriptase polymerase chain reaction (RT-PCR) and group II were 50 age and sex-matched healthy subjects as a control group. All patients followed the new direct-acting antiviral regimen (Sofosbuvir (400 mg) + Daclatasvir (60 mg) + Ribavirin (600 mg)) for 12 weeks.
All patients were subjected to:
Complete clinical history, thorough clinical examination and routine laboratory investigations including: Complete Blood Count (CBC), Liver function tests (AST, ALT and albumin). Hepatitis C virus antibody (anti-HCV) was detected in serum by ELISA, Hepatitis C virus RNA (HCV-RNA) was quantified by quantitative real-time polymerase chain reaction (PCR) before and 6, 12, 24 weeks after the end of treatment (EOT).
Liver stiffness measurement (LSM) by FibroScan was performed to assess the state of liver fibrosis. In addition, different serum non-invasive biomarkers of fibrosis were detected including, AST/ALT ratio (AAR), Fib-4 index and AST/platelet ratio index (APRI score).
Relative quantification of serum miR-122, miR-155, miR-196 and miR-29 was performed using quantitative real-time reverse transcription polymerase chain reaction (RT- qPCR) after microRNA extraction. A house keeping gene SNORD68 was used as internal control for relative quantification of miRNAs expression.
Interleukin 10 (IL-10) rs1800896 and rs1800872 SNP analysis was carried out by polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) assay.
Out of 150 HCV-4 patients, 141 (94%) achieved SVR12 to (SOF/DAV+RIB) regimen. Only 9 patients were resistant to the treatment (6%).
Regarding fibrosis state, 13.5 percent of sustained virological responders (SVR) belonged to stage 0, 59.6 percent to stage 1, 24.1 percent to stage 2, and 2.8 percent to stage 3. Among non-responders, 100% of the patients belonged to stages 2 and 3, with 44.4 percent in stage 2 and 55.6 percent in stage 3.
A significant difference was reported between patients and controls regarding miRNAs level where the levels of serum miR-122 and miR-155 were significantly higher in the chronic hepatitis C (CHC) patients than in healthy controls, On the other hand, the levels of serum miR-196 and miR-29 were significantly lower in the chronic hepatitis C patients than in healthy controls.
Analyzing serum miRNAs profiles in HCV-patients regarding treatment response showed that the expression levels of miR-122 and miR-155 were significantly higher in non-responders to DAAs than in responders. In contrast, the expression levels of miR-29 and miR-196 were significantly higher in responders to DAAs than in non-responders. Receiver Operating characteristic (ROC) curve analysis revealed that all the studied miRNAs could serve as valuable biomarkers for prediction of response to DAAs.
We investigated whether there is any correlation between levels of our four-studied miRNAs with baseline HCV RNA levels; Bivariate correlation analysis revealed positive significant strong correlation between miRNAs 122, 155 and HCV RNA levels (r= 0.856, p <0.001), (r= 0.798, p <0.001), respectively. A negative significant strong and moderate correlation between miRNAs 29, 196 and HCV RNA levels was observed (r= -0.808, p <0.001), (r= -0.725, p <0.001) respectively.
Receiver Operating characteristic (ROC) curve analysis was performed to determine the diagnostic accuracy of miRNAs 122, 155, 196 and 29 as non-invasive biomarker for assessment of degree of liver fibrosis versus other serum indices (APRI, FIB-4 and AAR); it revealed that APRI score had the highest diagnostic accuracy for assessment of liver fibrosis followed by FIB-4 index, miR-155, then both miR-122 and miR-29 and finally miR-196. AAR gave non-significant results with poor diagnostic accuracy in our findings.
Our study analyzed IL-10 promoter SNPs: -1082 G/A and - 592 C/A and observed that “C- allele” of IL- 10 (-592 C/A) is significantly higher in HCV infected patients compared to controls. However, IL-10 (-1082 G/A) genotypes showed no significant difference between patients and controls.
Regarding the response to DAAs, the current study stated that no significant difference between IL-10 (-1082 G/A) or IL 10 (-592 C/A) among responders and non-responders patients.