Hepatitis C virus NS5A and core proteins induce oxidative stress-mediated calcium signalling alterations in hepatocytes

Modulation of PI3K-LXRα-dependent lipogenesis mediated by oxidative/nitrosative stress contributes to inhibition of HCV replication by quercetin

“…[4][5][6][7] In the present research, quercetin was able to significantly inhibit HCV-induced ROS and RNS formation in HCV-replicating cells through its antioxidant activity. It has been also described that HCV replication induces lipid peroxidation that in turn reduces the amount of HCV RNA.…”

“…Replication studies were carried out using a SYBR Green kit (Roche Diagnostics GmbH) and two specific primer sets (5 0 -CCTGTGAGGAA CTACTGTCT-3 0 and 5 0 -CTATCAGGCAGTACCACAAG-3 0 for HCV, spanning 255 nucleotides of the 5 0 non-translated region; 5 0 -AAAGCCGCTCGCAAGAGTGCG-3 0 and 5 0 -ACTT GCCTCCTGCAAAGCAC-3 0 for histone H3). 6 The number of HCV RNA copies was determined by crossing point interpolation into standard curves, which were generated by reverse transcription of serially diluted, in vitro synthesized viral RNA (genomic) followed by quantitative PCR. The total amount of RNA per reaction was kept constant (1 mg) by the addition of Huh7 RNA.…”

“…3 Although the molecular mechanisms of HCV pathogenesis remain unclear, oxidative/nitrosative stress associated with reactive oxygen and nitrogen species (ROS/RNS) generation is emerging as a key step and a major initiator in the development and the progression of liver damage. [4][5][6][7] Hepatic steatosis is often observed in patients with chronic hepatitis C and has been reported to be associated with insulin resistance and progression of fibrosis in the liver. 8,9 1 The molecular mechanisms of HCV-associated steatosis are poorly understood, 10 but they are mediated in large part by changes in lipogenic and oxidative stress-mediated proinflammatory cytokines gene expression associated with HCV proteins.…”

Liver X receptor α-mediated regulation of lipogenesis by core and NS5A proteins contributes to HCV-induced liver steatosis and HCV replication

“…15 We have previously described that NS5A and core proteins induce oxidative stress-mediated Ca 2 þ homeostasis alterations in CHL, which might underlie the effects of both proteins on the PI3K/AKT pathway. 31 It has been proposed that the PI3K/AKT pathway activation is required for HCV core-mediated SREBP-1 induction, 46 process synergistically supported by HCV NS4B protein. 15,49 Thus, AKT has been shown to be able to increase endoplasmic reticulum to Golgi transport of SREBP and mature SREBP levels and SREBP phosphorylation, whereas the effects on SREBP transcription has not been observed.…”

“…Replication studies were carried out using a SYBR Green kit (Roche Diagnostics GmbH) and two specific primer sets (5 0 -CCTGTGAGGAAC TACTGTCT-3 0 and 5 0 -CTATCAGGCAGTACCACAAG-3 0 for Hepatitis C virus-associated steatosis MV García-Mediavilla et al HCV, spanning 255 nucleotides of the 5 0 nontranslated region; 5 0 -AAAGCCGCTCGCAAGAGTGCG-3 0 and 5 0 -ACTTGCCT CCTGCAAAGCAC-3 0 for histone H3). 31 The number of HCV RNA copies was determined by crossing point interpolation into standard curves, which were generated by reverse transcription of serially diluted, in vitro synthesized viral RNA (genomic) followed by quantitative PCR. The total amount of RNA per reaction was kept constant (1 mg) by the addition of Huh7 RNA.…”