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. 2020 Nov-Dec;34(6):3301-3308.
doi: 10.21873/invivo.12168.

Additive Effects of Zinc Chloride on the Suppression of Hepatitis A Virus Replication by Interferon in Human Hepatoma Huh7 Cells

Tatsuo Kanda  1 , Reina Sasaki  2 , Ryota Masuzaki  2 , Hiroshi Takahashi  2 , Mariko Fujisawa  2 , Naoki Matsumoto  2 , Hiroaki Okamoto  3 , Mitsuhiko Moriyama  2
Affiliations

Additive Effects of Zinc Chloride on the Suppression of Hepatitis A Virus Replication by Interferon in Human Hepatoma Huh7 Cells

Tatsuo Kanda et al. In Vivo. 2020 Nov-Dec.

Abstract

Background/aim: Hepatitis A virus (HAV) infection is still one of the serious health problems worldwide, despite the existence of effective vaccines for HAV. Zinc compounds have antiviral activities against various DNA and RNA viruses. Therefore, we investigated the effects of zinc compounds on the antiviral activity of interferon against HAV.

Materials and methods: The effects of zinc compounds with or without interferon on HAV genotype IIIA HA11-1299 replication were examined in human hepatoma Huh7 cells. Cell viability was examined by the MTS assay. Inflammasome associated gene expression was examined by real-time reverse transcription-polymerase chain reaction.

Results: Both zinc sulfate and zinc chloride had an inhibitory effect on HAV replication. Zinc sulfate tended to enhance while zinc chloride significantly enhanced the anti-HAV effect induced by interferon-alpha-2a. Zinc chloride significantly up-regulated mitogen-activated protein kinase 12 (MAPK12) and down-regulated 6 related genes [baculoviral IAP repeat containing 3 (BIRC3), interleukin 1 beta (IL1B), proline-serine-threonine phosphatase interacting protein 1 (PSTPIP1), prostaglandin-endoperoxide synthase 2 (PTGS2), PYD and CARD domain containing (PYCARD), and tumor necrosis factor (TNF)].

Conclusion: Zinc chloride inhibits HAV replication and has additive effects on the anti-HAV activities of interferon.

Keywords: Antivirals; HAV; inflammasomes; interferon; zinc.

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Conflict of interest statement

No competing interests exist in relation to this study.

Figures

Figure 1
Figure 1. Illustration of the protocols for hepatitis A virus (HAV) infection, treatment with zinc compounds and cellular RNA extraction. PBS: Phosphate-buffered saline; FCS: fetal calf serum; DMEM: Dulbecco’s Modified Eagle Medium; IFN: interferon.
Figure 2
Figure 2. Effects of zinc compounds on the viability of human hepatocytes. The viability of HepG2 and Huh7 cells treated with zinc compounds for 72 h. (A) Zinc chloride (ZnCl2), (B) zinc sulfate (ZnSO4). Cell viability was measured by MTS assay (see Materials and Methods section). Data are expressed as means±standard deviations of triplicate determinations from 3 independent experiments.
Figure 3
Figure 3. Zinc compounds and interferon inhibit hepatitis A virus (HAV) replication. Drugs were added 24h post infection with 0.01 MOI HAV genotype IIIA HA11-1299. HAV RNA was measured by real-time reverse transcription-polymerase chain reaction. IFN: Interferon. Data are expressed as means±standard deviations of triplicate determinations from 3 independent experiments.
Figure 4
Figure 4. Zinc compounds enhanced the antiviral activity of interferon against hepatitis A virus (HAV) replication. (A) Zinc chloride (ZnCl2), (B) zinc sulfate (ZnSO4). Drugs were added 24h post infection with 0.01 MOI HAV genotype IIIA HA11-1299. HAV RNA was measured by real-time reverse transcription-polymerase chain reaction. IFN: Interferon. Data are expressed as means±standard deviations of triplicate determinations from 3 independent experiments.
Figure 5
Figure 5. Clustergram of gene expression analysis of inflammasome associated signaling pathways in human hepatoma Huh7 cells treated with or without zinc chloride (Zn Cl2). C: Untreated control.
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