Antiviral activity and mechanism of pinecone extract of pinus yunnanensis against Lassa virus infection
10.3969/j.issn.1002-2694.2024.00.145
- VernacularTitle:云南松松塔粗提物体外抗拉沙病毒活性及机制的研究
- Author:
Zhen-Lin LIU
1
;
Man-Lin ZHANG
;
Qi LIU
;
Cong WANG
Author Information
1. 大理大学基础医学院病原生物学综合实验室,大理 671000
- Keywords:
pine cone extract of pinus yun nanensis;
Lassa virus;
antiviral activity;
antiviral mechanism
- From:
Chinese Journal of Zoonoses
2024;40(10):905-912
- CountryChina
- Language:Chinese
-
Abstract:
A Lassa pseudovirus system and Lassa cell-cell fusion were used to detect the anti-LASV activity of Pineta crude extract.The Lassa pseudovirus system was constructed with the lentivirus skeleton plasmid PNL-4-3.Luc.R-E.After transfec-tion into 293T cells with Lassa-coated protein granules,a fake Lassa virus containing luciferase reporter gene was generated.The anti-LASV activity of pine tar crude extract was detected in a BSL-2 biosafety laboratory.Second,a Lassa cell-cell fusion model was constructed by enzymatically cutting,linking,and identifying existing plasmids to obtain recombinant plasmids,which were transfected into 293T cells to obtain fluorescent cells with Lassa envelope protein on the surface.These cells were mixed with Huh-7 target cells in 96-well plates,and membrane fusion occurred at 37 ℃ under acidic conditions.Subsequently,a membrane fusion system was used to detect the anti-LASV activity of pine tar crude extract.The specific period of action of pine tar crude extract on LASV pseudovirus infection was analyzed,and the antiviral target of pine tar crude extract was an-alyzed with time of addition and time of removal tests.The protein content of p24 was detected by western blotting to determine the viral cleavage effect of pine tar crude extract.The crude ex-tract of pine tar effectively inhibited LASV infection.Four lineages of LASV pseudoviruses(LASV lineages Ⅰ,Ⅱ,Ⅲ,and Ⅳ)were used to infect target cells,and the IC50 values were as follows:42.06 μg/mL,22.26 μg/mL,80.57 μg/mL,and 19.32 μg/mL inhibited the cell-cell fusion activity mediated by LASV envelope protein(GPC),and the IC50 values were as fol-lows:122.9 μg/mL,64.43 μg/mL,90.05 μg/mL and 195.6 μg/mL.The crude extract of Songta had broad-spectrum antiviral effects,and effectively inhibited infection with arenavirus,including lymphocytic choriomeningitis virus,Luho virus,and Junin virus pseudovirus.The IC50 values were 2.72 μg/mL,12.02 μg/mL,and 29.95 μg/mL,respectively.The crude extract of pine tar acted in early stages of LASV infection.The inhibitory rate after was significantly higher with addition of crude extract within 2 h rather than 4 h after infection,and the adsorption and fusion of LASV pseudovirus were largely blocked.The crude extract of pine tar had no cleavage effect on LASV pseudovirus,and no significant difference in p24 protein content was ob-served between the treated group and the untreated group.This study confirmed that crude Yunnan pine tree extract effectively inhibits LASV infection in vitro,and has broad-spectrum antiviral effects.The experimental results showed that crude pine tar extract acts mainly in early stages of viral infection,by blocking viral adsorption and fusion.These findings provide a reference for further research on the inhibitory effects of pinecone extract toward LASV.
