Antiviral Activity of Chlorophyll Extracts from Tetraselmis sp., a Marine Microalga, Against Zika Virus Infection
Abstract
:1. Introduction
2. Results and Discussion
2.1. Inhibitory Effects of Tetraselmis sp. Acetone Extracts on ZIKV
2.2. Effect of T90A on Interferon Response in ZIKV-Infected Cells
2.3. Isolation and Purification of Main Compounds from T90A
2.4. Characterization of Four Main Compounds from T90A
2.5. Inhibitory Effects of Chlorophylls from T90A on ZIKV Infection
2.6. Molecular Docking Analysis of Chlorophylls to ZIKV Proteins
3. Materials and Methods
3.1. Mass Production of Tetraselmis sp.
3.2. Materials and Reagents
3.3. Preparation of Extracts and Purification of Main Compounds
3.4. Structural Identification of Compounds
3.5. Cell Culture
3.6. Measurement of Cytotoxicity and Calculation of SI
3.7. Measurement of Viral Plaque Formation Rate
3.8. Measurement of mRNA Expression Levels
3.9. Measurement of Virucidal Effect
3.10. Molecular Docking Analysis between Chlorophylls and ZIKV Proteins
3.11. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antiviral Activity | Virucidal activity | Cytotoxicity | ||||
---|---|---|---|---|---|---|
Pre-Treatment | Post-Treatment | |||||
EC50 (μg/mL) | SI | EC50 (μg/mL) | SI | EC50 (μg/mL) | SI | CC50 (μg/mL) |
36.4 ± 0.5 | 21.1 | 31.3 ± 0.8 | 24.6 | 34.1 ± 0.2 | 22.6 | 769.6 ± 6.9 |
Chlorophyll | Protein | Binding energy (kcal/mol) |
---|---|---|
Chlorophyll a | Envelope protein | −276.459 |
NS2B/NS3 | −767.837 | |
RdRp | −692.708 | |
Chlorin e6-131-152-dimethyl-173-phytyl ester | Envelope protein | −374.144 |
NS2B/NS3 | −779.602 | |
RdRp | −727.470 |
Chlorophyll | Protein | Non-Bond Interaction in Active Site |
---|---|---|
Chlorophyll a (C1) | Envelope protein | Mg2+-TRP101, Mg2+-GLY102, C1-LEU107 (2), C1-TRP101 (2) |
NS2B/NS3 | Mg2+-ASN1152, Mg2+- GLY1153, Mg2+-VAL1154, Mg2+-VAL1162, C1-VAL1036, C1-HIS1051 (3), C1-ALA1132 (3), C1-SER1135 (2), C1-TYR1150, C1-GLY1151, C1-ASN1152 (2), C1-TYR1161 (4) | |
RdRp | Mg2+-GLU707 (2), Mg2+-TRP848, C1-VAL708, C1-HIS713, C1-HIS732 (2), C1-CYS849, | |
Chlorin e6-131-152-dimethyl-173-phytyl ester (C2) | Envelope protein | Mg2+-GLY104, Mg2+-Cys105, Mg2+-GLY106 C2-TRP101(5), C2-GLY104, C2-GLY106, C2-LEU107 |
NS2B/NS3 | Mg2+-ASP1129 (2), Mg2+-ASN1152, Mg2+-GLY1153, Mg2+-VAL1162, C2-HIS1051, C2-ALA1132, C2-VAL1154, C2-TYR1161 (2) | |
RdRp | Mg2+-GLU707, C2-TRP702 (2), C2-VAL708 (2), C2-TRP848 (2) |
Gene | Sequence | Primer |
---|---|---|
ZIKV NS1 | 5’-CRA CTA CTG CAA GYG GAA GG-3’ | F |
5’-GCC TTA TCT CCA TTC CAT ACC-3’ | R | |
Monkey GAPDH | 5’-GCA AAT TCC ATG GCA CCG T-3’ | F |
5’-TCG CCC CAC TTG ATT TTG G-3’ | R | |
Monkey IFNγ | 5’-CGA ATG TCC AAC GCA AAG CAG TAC-3’ | F |
5’-TGC TCT TCG ACC TCG AAA CAT CTG-3’ | R | |
Monkey IFIT1 | 5’- GGA TTC TGT ACA ATA CAC TAG AAA CCA-3’ | F |
5’- CTT TTG GTT ACT TTT CCC CTA TCC-3 ‘ | R | |
Monkey IFIT2 | 5’- ATC CCC CAT CGC TTA TCT CT-3’ | F |
5’- CCACCTCAATTAATCAGGCACT-3’ | R |
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Kang, N.; Kim, E.-A.; Park, A.; Heo, S.-Y.; Heo, J.-H.; Lee, W.-K.; Ryu, Y.-K.; Heo, S.-J. Antiviral Activity of Chlorophyll Extracts from Tetraselmis sp., a Marine Microalga, Against Zika Virus Infection. Mar. Drugs 2024, 22, 397. https://doi.org/10.3390/md22090397
Kang N, Kim E-A, Park A, Heo S-Y, Heo J-H, Lee W-K, Ryu Y-K, Heo S-J. Antiviral Activity of Chlorophyll Extracts from Tetraselmis sp., a Marine Microalga, Against Zika Virus Infection. Marine Drugs. 2024; 22(9):397. https://doi.org/10.3390/md22090397
Chicago/Turabian StyleKang, Nalae, Eun-A Kim, Areumi Park, Seong-Yeong Heo, Jun-Ho Heo, Won-Kyu Lee, Yong-Kyun Ryu, and Soo-Jin Heo. 2024. "Antiviral Activity of Chlorophyll Extracts from Tetraselmis sp., a Marine Microalga, Against Zika Virus Infection" Marine Drugs 22, no. 9: 397. https://doi.org/10.3390/md22090397