| Summary: | 碩士 === 國立臺灣海洋大學 === 食品科學系 === 99 === This study investigated the hot water extration of Chlorella
pyrenoidosa (Chlorella pyrenoidosa extract, CpE) for inhibiting influenza
A virus infection on in vitro and in vivo model. In vitro cell aaay, the
sequence of CpE treatment is according to the adsorption step of virus
infection; then divide into before virus adsorption (pre-treatment) and
after virus adsorption (post-treatment). We use cell viability assay (MTS
assay), cytopathic effect (CPE) assay, plaque assay, and viral RNA
expression (real-time PCR) to study the protective effects of CpE. The
beeter inhibition effect of CpE on influenza virus infection is during
pre-treatment stage, and CpE-2 has the best protective effect. The
concentration of CpE-1 and CpE-2 at more than 125 g/ml, the
anti-hemagglutination ability of CpE has 2 HAU (hemagglutinating unit).
CpE directly reacted the virus particles (virucidal) can effectively inhibit
the plaque formation, but CpE directly reacted cell (cell-treatment) has no
significant inhibitory effect. In virucidal treatment, 5 and 10 mg/ml of
CpE-2 show more than 95% of inhibition effect. Above all, the inhibition
mechanism of influenza virus infection of CpE is directly affecting on the
adsorption of influenza virus to host cell. In BALB/c mice animal model,
the 4 weeks-old mice infecte with lethal doses influenza virus and the
mortality rate is 100%. The mice feeded CpE-2 for a week and CpE-2
direct treated with virus (virucide) before infection, the mortality rate of
these mice are effectively reduced.
RNA expression of mouse lung were detected by real-time PCR,
influenza virus H1N1-NP, and inflammational cytokine: TNF-a, IL-1b,
2
IL-6, and antiviral cytokine: IFN-g, IL-12 in CpE-2 treated group were
lower than virus alone group, histological also showed less infiltration of
nertuophils, indicating CpE-2 treated can reduced virus infection and
inflammation of lung.
In summary, the Chlorella pyrenoidosa extract show a protective
effect against influenza virus infection on both in vitro cell and animal
experiment.
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