Monocyte infiltration in the brain of ICR suckling mice that received i.c. injection of various doses of ZIKV.

Six-day-old ICR suckling mice received 102 to 105 PFU ZIKV by i.c. or i.p. injection. Mice were sacrificed, and the brain tissues of mice that received ZIKV by (A) i.c. or (B) i.p. were collected and subjected to hematoxylin and eosin staining. The biopsy specimens were examined by microscope. (C, D) The quantitation of monocyte infiltration by IHC assay.</p

Survival rate, body weight, and clinical score of ICR suckling mice infected with ZIKV by i.p. and i.c. injection.

Six-day-old ICR suckling mice received 102 to 105 PFU ZIKV by i.c. (n = 5) or i.p. (n = 5) injection. (A–D) Survival rate, (E–H) clinical score, and (I–L) body weight recorded every day. Mice were sacrificed at 8 dpi. Disease severity was scored as follows: 0: healthy, 1: body weight loss and ruffled fur, 2: lethargy and unsteady gait, 3: kinetic tremors and severe ataxia, 4: paralysis, and 5: death.</p

ZIKV replication and ZIKV-induced monocyte infiltration were suppressed by Az treatment in ZIKV-infected ICR suckling mice.

Six-day-old ICR suckling mice were divided into four groups. Group 1 received heat inactivated ZIKV; group 2 received 10 mg/kg of Az but without ZIKV infection (Az 10 mg/kg); group 3 received activated ZIKV and saline treatment; group 4 received activated ZIKV and 1 mg/kg Az; and group 5 received activated ZIKV and 10 mg/kg Az. Mice were sacrificed at 7 dpi, and the brain tissues were collected for analysis of (A) ZIKV RNA replication, (B) viral titer, and (C) monocyte infiltration. (D) The quantitation of monocyte infiltration. *P < 0.05.</p

Survival rate, body weight, and clinical score in ZIKV-infected ICR suckling mice treated with Az.

Six-day-old ICR suckling mice were divided into four groups. Group 1 received heat inactivated ZIKV; group 2 received 10 mg/kg of Az but without ZIKV infection (Az 10 mg/kg); group 3 received activated ZIKV and saline treatment; group 4 received activated ZIKV and 1 mg/kg Az; and group 5 received activated ZIKV and 10 mg/kg Az. (A) Survival rate, (B) clinical score, and (C) body weight were recorded every day. Mice were sacrificed at 7 dpi. Disease severity was scored as follows: 0: healthy, 1: body weight loss and ruffled fur, 2: lethargy and unsteady gait, 3: kinetic tremors and severe ataxia, 4: paralysis, and 5: death. *P < 0.05.</p

ZIKV replication in different organs of mice that underwent i.p. and i.c. injection of various doses of ZIKV.

Six-day-old ICR suckling mice received 102 to 105 PFU ZIKV by i.c. or i.p. injection. Mice were sacrificed, and the brain, liver, spleen, and kidney tissues were collected to analyze ZIKV RNA replication and protein synthesis. (A and B) ZIKV protein synthesis in the brain tissues of mice that received ZIKV by (A) i.c. or (B) i.p. as detected by Western blot analysis with anti-ZIKV NS2B antibody. GAPDH served as equal loading control. (C and D) ZIKV RNA levels in the brain tissue of mice that received ZIKV by (C) i.c. or (D) i.p. as determined by qRT-PCR analysis. The ZIKV RNA level was normalized by the cellular gapdh mRNA level. (E and F) ZIKV viral titer in the brain tissue of mice that received ZIKV by (E) i.c. or (F) i.p. as determined by plaque forming assay. (G and H) ZIKV RNA levels in the liver tissue of mice that received ZIKV by (G) i.c. or (H) i.p. as determined by qRT-PCR analysis. (I and J) ZIKV RNA levels in the spleen tissue of mice that received ZIKV by (I) i.c. or (J) i.p. as determined by qRT-PCR analysis. (K and L) ZIKV RNA levels in the kidney tissue of mice that received ZIKV by (K) i.c. or (L) i.p. as determined by qRT-PCR analysis.</p

ZIKV replication in the brain of mice i.p. injected with 10⁴ PFU ZIKV.

Six-day-old ICR suckling mice received 104 PFU ZIKV by i.p. injection, and brain tissues were collected every day (n = 3) for analysis of ZIKV (A) protein synthesis, (B) RNA replication, (C) viral titer, and (D) monocyte infiltration. (E) The quantitation of monocyte infiltration by IHC assay. *P P0.01.</p

Sulforaphane Suppresses Hepatitis C Virus Replication by Up-Regulating Heme Oxygenase-1 Expression through PI3K/Nrf2 Pathway

<div><p>Hepatitis C virus (HCV) infection-induced oxidative stress is a major risk factor for the development of HCV-associated liver disease. Sulforaphane (SFN) is an antioxidant phytocompound that acts against cellular oxidative stress and tumorigenesis. However, there is little known about its anti-viral activity. In this study, we demonstrated that SFN significantly suppressed HCV protein and RNA levels in HCV replicon cells and infectious system, with an IC₅₀ value of 5.7 ± 0.2 μM. Moreover, combination of SFN with anti-viral drugs displayed synergistic effects in the suppression of HCV replication. In addition, we found nuclear factor erythroid 2-related factor 2 (Nrf2)/HO-1 induction in response to SFN and determined the signaling pathways involved in this process, including inhibition of NS3 protease activity and induction of IFN response. In contrast, the anti-viral activities were attenuated by knockdown of HO-1 with specific inhibitor (SnPP) and shRNA, suggesting that anti-HCV activity of SFN is dependent on HO-1 expression. Otherwise, SFN stimulated the phosphorylation of phosphoinositide 3-kinase (PI3K) leading Nrf2-mediated HO-1 expression against HCV replication. Overall, our results indicated that HO-1 is essential in SFN-mediated anti-HCV activity and provide new insights in the molecular mechanism of SFN in HCV replication.</p></div

Anti-HCV activity and cytotoxicity of SFN and its analogs.

Anti-HCV activity and cytotoxicity of SFN and its analogs.</p

HCV NS5A Up-Regulates COX-2 Expression via IL-8-Mediated Activation of the ERK/JNK MAPK Pathway

<div><p>Chronic hepatitis C virus (HCV) infection leads to intrahepatic inflammation and liver cell injury, which are considered a risk factor for virus-associated hepatitis, cirrhosis, and hepatocellular carcinoma worldwide. Inflammatory cytokines are critical components of the immune system and influence cellular signaling, and genetic imbalances. In this study, we found that cyclooxygenase-2 (COX-2) and interleukin-8 (IL-8) were significantly induced by HCV infection and HCV NS5A expression, and induction of COX-2 correlated with HCV-induced IL-8 production. We also found that the ERK and JNK signaling pathways were involved in the regulation of IL-8-mediated COX-2 induction in response to HCV infection. Using a promoter-linked reporter assay, we identified that the C/EBP regulatory element within the COX-2 promoter was the dominant factor responsible for the induction of COX-2 by HCV. Silencing C/EBP attenuated HCV-induced COX-2 expression. Our results revealed that HCV-induced inflammation promotes viral replication, providing new insights into the involvement of IL-8-mediated COX-2 induction in HCV replication.</p></div

Anti-HCV activity of SFN in combination with antiviral drugs in Ava5 cells.

<p>Anti-HCV activity of SFN in combination with antiviral drugs in Ava5 cells.</p