In Egypt, about 9 million people are infected with HCV in 2010. The complex relationship of INFs, miRNAs and HCV infection was undetermined and needs to be clarified. The current work was aimed to study the effect of combination therapy (pegylated interferon alpha-2b plus ribavirin) on microRNA-128 and microRNA-296 in patients with chronic hepatitis C. Also to verify that; these 2 types of microRNAs might be useful to differentiate between responders and non responders either in the pre- treatment or post- treatment group.
16 male subjects were selected from Hepatic Center, Benha Fever Hospital, in addition to 10 healthy volunteers who served a as control group. Their ages ranged from 23-56 years (y) with mean value 35.65 ± 9.67 y. The patients received doses of combination therapy including peginterferon alfa-2b (1.5µg /kg/week) plus weight-based ribavirin (10.6 mg/kg per day). The subjects were categorized into 2 groups:
1) Pre-treatment group: Comprised 16 patients with chronic HCV infection.
2) Post-treatment group: patients were subdivided into two subgroups:
• Subgroup A: included 10 responders.
• Subgroup B: included 6 non-responders .
All subjects were subjected to the followings:
(A) Non-laboratory investigations:
1) Full history taking.
2) Complete physical examination.
3) Abdominal ultrasonography scanning.
4) ECG.
5) Fundus examination.
(B) Laboratory investigations:
Ι- Biochemical investigations:
1. Complete Blood Count (CBC) (Smith and Lipworth, 1995).
2. Liver function tests :
* Serum AST, ALT (Reitman and Frankel, 1957).
* Serum alkaline phosphatase (Teitz and Shuey, 1986).
* Serum albumin (Dumas et al., 1971)
* Serum Total bilirubin (Malloy and Evelyn, 1937)
* Plasma prothrombine time (Hirsh et al., 1992).
* Serum Alpha feto-protein (Stuart et al., 1996).
3. Fasting serum glucose (Trinder, 1969).
4. Serum TSH level (Burger and Patel, 1977).
5. Serum creatinine (Henry, 1974).
6. Serum ANA titre (Notman et al., 1975).
7. Bilharzial antibodies by IHA (Azab and El Zayat, 1996).
8. HCV antibodies by enzyme immunoassay (Arash et al., 1993).
9. Hepatitis B markers as hepatitis B surface antigen (HBsAg) by enzyme immunoassay (Blumberg, 1971).
10. Quantitative hepatitis C virus RNA PCR (before and 12 weeks after combination therapy) (Livak and Schittgen, 2001).
11. Extraction and molecular analysis of circulating miR-128 and miR-296-5p (before and 12 weeks after combination therapy) by real time PCR (Livak and Schittgen, 2001).
ΙΙ- Histopathological investigations: biopsy was done only for HCV patients.
The results revealed that miR-128 and miR-296 were expressed in whole blood collected from healthy individuals (Table: 8& fig. 29, 30).
Furthermore, miR-128 showed significant decrease while mir-296 showed non significant decrease in the pre- treatment group compared to the control group (Table: 21 & fig. 29, 30).
Moreover, Our results showed non significant increase in expression of miR-128 while miR-296 showed significant decrease in the post treatment compared either with pre-treatment group (Table: 23 & fig. 29, 30) or with control group (Table: 22 & fig. 29, 30)
Comparative studies of responders versus non responders showed non significant differences either in the pre-treatment (Table: 25& fig.31, 32) or in the post-treatment groups (Table: 26 & fig. 33, 34).
CONCLUSION
It could be concluded that the levels of expression of miR-128 and miR-296 were affected by chronic hepatitis C as well as the combination therapy. Until now, miRNAs studied were not useful indicators to differentiate between responders and non responders to combination therapy either in the pre-treatment or post-treatment group.
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