The enhancing impact of amino termini of Hepatitis C virus core protein on activation of Hepatic Stellate Cells
Gastroenterology and Hepatology from Bed to Bench,
Vol. 13 No. 1 (2020),
22 January 2020
,
Page 57-63
https://doi.org/10.22037/ghfbb.v13i1.1668
Abstract
Background: The core protein is recognized as the most important fibrosis inducer of Hepatitis C virus (HCV). While the exogenous fibrotic effect of HCV core protein has been reported previously, the endogenous effect and role of two termini is yet to be investigated.
Aim: To study the potential effect of carboxyl and amino termini of HCV core protein on HSCs activation.
Methods: Plasmids expressing full length, carboxyl-truncated(T1) or amino-truncated(T3) versions of the core were transfected into LX?2 cells. MTT assay was performed to evaluate the cytotoxicity of the endogenous expression of different regions of core protein on these cells. Afterwards, the total RNA was reverse transcribed and introduced into quantitative polymerase chain reaction(qPCR) to measure the expression level of collagen type I (COL1A1), ?-smooth muscle actin (a-SMA), tissue metalloproteinase inhibitor?1 (TIMP-1) and transforming growth factor-?1 (TGF-?1). In addition, TGF-?1 as a fibrotic factor was also assayed in the supernatant of LX-2 cells using ELISA method.
Results: The full and T1 versions of the core exhibited a measurable proliferative effect on LX?2 cells (P<0.05). Analysis of the gene expression was also showed that in spite of amino-truncated version these constructs represented a significant activation impact when compared to the empty plasmid. Moreover, the result of TGF?? assay was also in agreement with the results of mRNA expression analysis.
Conclusion: The endogenous expression of the full and carboxyl-truncated versions of the core exhibited a significant activator effect on HSCs. Therefore, it can be concluded that amino domain of HCV core protein performs a stellate cell activation role.
- Hepatitis C virus
- hepatic stellate cell
- core protein
- liver fibrosis
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