连翘脂苷A通过调控TLR3-NF-κB信号通路抑制H9N2亚型禽流感病毒复制

doi:10.11843/j.issn.0366-6964.2025.10.045

摘要/Abstract

摘要：

本研究旨在探讨连翘脂苷A(forsythiaside A, FTA)对H9N2亚型禽流感病毒(H9N2 AIV)在肺微血管内皮细胞(pulmonary microvascular endothelial cells, PMVECs)中复制的抑制作用及其潜在机制。采用不同浓度的连翘脂苷A处理H9N2 AIV感染的PMVECs，使用MTT法检测细胞毒性，采用RT-qPCR分析病毒复制情况。通过Western blot检测TLR3、TRIF和NF-κB蛋白表达水平，采用ELISA检测炎性细胞因子(TNF-α、IL-6)及Ⅰ型干扰素(IFN-α、IFN-β)的分泌情况。研究结果表明，连翘脂苷A能显著抑制H9N2 AIV的复制(P < 0.05)，其抑制作用呈剂量依赖性；下调TLR3、TRIF及NF-κB蛋白的过表达(P < 0.05)，降低促炎性细胞因子分泌(P < 0.05)，同时增强Ⅰ型干扰素的生成(P < 0.05)。连翘脂苷A通过调控TLR3-NF-κB信号通路及细胞因子表达，展现出针对H9N2 AIV的显著抗病毒活性，具有作为新型宿主靶向抗病毒药物的潜力。

关键词: 连翘脂苷A, H9N2禽流感病毒, TLR3-NF-κB信号通路, 细胞因子, 肺微血管内皮细胞, 抗病毒治疗, 中药

Abstract:

This study aimed to investigate the inhibitory effects of Forsythiaside A (FTA) on the replication of H9N2 avian influenza virus (H9N2 AIV) in pulmonary microvascular endothelial cells (PMVECs) and explore its underlying mechanisms. PMVECs infected with H9N2 AIV were treated with varying concentrations of FTA. Cytotoxicity was assessed using the MTT assay, and viral replication was analyzed via RT-qPCR. Protein expression levels of TLR3, TRIF, and NF-κB were evaluated using Western blot, while cytokine secretion (TNF-α, IL-6) and type Ⅰ interferons (IFN-α, IFN-β) were measured through ELISA. The results showed that FTA significantly inhibited the replication of H9N2 AIV (P < 0.05) in a dose-dependent manner. It downregulated the overexpression of TLR3, TRIF, and NF-κB proteins (P < 0.05), suppressed the secretion of pro-inflammatory cytokines (P < 0.05), and enhanced the production of type Ⅰ interferons (P < 0.05). FTA exhibits significant antiviral activity against H9N2 AIV by modulating the TLR3-NF-κB signaling pathway and regulating cytokine expression, highlighting its potential as a novel host-targeted antiviral agent.

Key words: forsythiaside A, H9N2 avian influenza virus, TLR3-NF-κB signaling pathway, cytokines, pulmonary microvascular endothelial cells, antiviral therapy, traditional Chinese medicine

中图分类号:

S853.74

张泰铭, 李雅萱, 汪长寿, 胡格. 连翘脂苷A通过调控TLR3-NF-κB信号通路抑制H9N2亚型禽流感病毒复制[J]. 畜牧兽医学报, 2025, 56(10): 5266-5276.

ZHANG Taiming, LI Yaxuan, WANG Changshou, HU Ge. Forsythiaside A Inhibits Replication of H9N2 Avian Influenza Virus by Modulating the TLR3-NF-κB Signaling Pathway[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 5266-5276.

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基因名称 Gene name	序列(5′→3′)Sequence
基因名称 Gene name	正向引物Forward primer	反向引物Reverse primer
TLR3	AGCCATTTGAAGCTGACAC	CTGCATCCAAGAGAGCAAG
TRIF	CTGTCATTTCTTGAGCGTGA	ATCCAGGTTATTGCTTCTGTG
NF-κB	CCCCTGAGAAAGAAACACA	CCTGAAACCCCACATCC
GAPDH	ATGGCTACAGCAACAGGGT	TTATGGGGTCTGGGATGG

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