Silencing of α-complex protein-2 reverses alcohol- and cytokine-induced fibrogenesis in hepatic stellate cells

Abstract

Background and aim α-complex protein-2 (αCP2) encoded by the poly (rC) binding protein 2(PCBP2) gene is responsible for the accumulation of type I collagen in fibrotic livers. In this study, we silenced the PCBP2 gene using a small interfering RNA (siRNA) to reverse alcohol-and cytokine-induced profibrogenic effects on hepatic stellate cells (HSCs). Methods Primary rat HSCs and the HSC-T6 cell line were used as fibrogenic models to mimic the initiation and perpetuation stages of fibrogenesis, respectively. We previously found that a PCBP2 siRNA, which efficiently silences expression of αCP2, reduces the stability of type I collagen mRNA. We investigated the effects of the PCBP2 siRNA on cell proliferation and migration. Expression of type I collagen in HSCs was analyzed by quantitative real-time PCR and western blotting. In addition, we evaluated the effects of the PCBP2 siRNA on apoptosis and the cell cycle. Results PCBP2 siRNA reversed multiple alcohol- and cytokine-induced profibrogenic effects on primary rat HSCs and HSC-T6 cells. The PCBP2 siRNA also reversed alcohol- and cytokine-induced accumulation of type I collagen as well as cell proliferation and migration. Moreover, the combination of LY2109761, a transforming growth factor-β1 inhibitor, and the PCBP2 siRNA exerted a synergistic inhibitive effect on the accumulation of type I collagen in HSCs. Conclusions Silencing of PCBP2 using siRNA could be a potential therapeutic strategy for alcoholic liver fibrosis.

Discovery of low-molecular weight anti-PD-L1 peptides for cancer immunotherapy

“…After incubation with the anti-PD-L1 peptides (5 μM) or anti-PD-L1 antibody (1 μM) for 24 h, Jurkat T cells were harvested from the supernatant. Proliferation of Jurkat cells was determined using the CellTiter-Glo luminescent cell viability assay (Promega, WI), and apoptosis of the cells were determined using the Dead Cell Apoptosis Kit with Annexin V Alexa Fluor® 488 and Propidium Iodide (Thermo-Fisher Scientific, Pittsburgh, PA) as we described before [21].…”

Development of a peptide-modified siRNA nanocomplex for hepatic stellate cells

“…Cytotoxicity of the sRNA nanocomplex in HSC-T6 cells was evaluated using MTT assay as described. 22,23 HSC-T6 cells seeded in 96-well plate (2500 cells/well) were incubated with SNCP, SNVP and SNPP nanocomplexes at 37 °C for 24 and 48h, followed by adding MTT to measure cytotoxicity.…”

Targeted Drug Delivery to Hepatic Stellate Cells for the Treatment of Liver Fibrosis

“…A wide range of therapeutic agents including small molecules, proteins, small interfering RNA (siRNAs), microRNA (miRNAs), and plasmids have been investigated for the treatment of liver fibrosis (Tu et al, 2014; Wang et al, 2015; Yoon et al, 2016; Liu et al, 2017; Middleton et al, 2018). However, antifibrotic agents cannot easily reach HSCs in fibrotic liver because HSCs only account for 5% to 8% of total liver cells (Geerts, 2001).…”