School of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.
College of Basic Medicine, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Biomed Res Int. 2022 Feb 15;2022:1040129. doi: 10.1155/2022/1040129. eCollection 2022.
Traditional Chinese medicine Yinhuapinggan granule (YHPG) has been used for treating upper respiratory tract infection like influenza, cough, and viral pneumonia. However, its active ingredients that really exert the main efficacy have not been well elucidated. This study is aimed at screening its antiviral components and investigating the potential therapeutic mechanisms of YHPG against the influenza A/PR8/34 (H1N1) virus in Madin Darby canine kidney (MDCK).
MDCK cells were infected with the influenza virus and then treated with ribavirin, YHPG, and main active ingredients in YHPG. Based on the maximum nontoxic concentration (TC), half-maximal toxic concentration (TC), half-maximal inhibitory concentration (IC), and therapeutic index (TI), interferon- (IFN-) and interleukin-6 (IL-6) levels were measured using enzyme-linked immunosorbent assay (ELISA), and the gene expression of TLR7, MyD88, tumor necrosis factor receptor-associated factor 6 (TRAF6), c-Jun amino terminal kinase (JNK), p38 mitogen-activated protein kinase (p38 MAPK), and p65 nuclear transcription factor-kappa B (p65 NF-B) was quantified using reverse transcription-polymerase chain reaction (RT-PCR).
The results indicated that the components of YHPG, such as ephedrine hydrochloride, pseudoephedrine hydrochloride, chlorogenic acid, and emodin, had significant antiviral effects. High and medium doses of YHPG effectively reduced the cytopathic effect (CPE) and significantly decreased IFN- and IL-6 levels in the supernatant. Simultaneously, the transcript levels of TLR7, MyD88, TRAF6, JNK, p38 MAPK, and p65 NF-B decreased in infected MDCK cells. Moreover, a certain dose-dependent relationship among different groups of YHPG was observed.
These results indicated that YHPG and the components of YHPG had a significant inhibitory function on the proliferation of the H1N1 virus. The mechanism might be associated with suppressing the activation of the TLR7/MyD88 signaling pathway, a decrease in the mRNA expression of key target genes, and inhibition of IFN- and IL-6 secretion.
中药银花平感颗粒（YHPG）已用于治疗上呼吸道感染，如流感、咳嗽和病毒性肺炎。然而，其发挥主要疗效的活性成分尚未得到很好的阐明。本研究旨在筛选其抗病毒成分，并探讨 YHPG 对甲型流感病毒/PR8/34（H1N1）在 Madin Darby 犬肾（MDCK）中的潜在治疗机制。
用流感病毒感染 MDCK 细胞，然后用利巴韦林、YHPG 和 YHPG 中的主要活性成分进行处理。根据最大无毒浓度（TC）、半最大毒性浓度（TC）、半最大抑制浓度（IC）和治疗指数（TI），采用酶联免疫吸附试验（ELISA）测定干扰素-（IFN-）和白细胞介素-6（IL-6）水平，并用逆转录-聚合酶链反应（RT-PCR）定量测定 TLR7、MyD88、肿瘤坏死因子受体相关因子 6（TRAF6）、c-Jun 氨基末端激酶（JNK）、p38 丝裂原活化蛋白激酶（p38 MAPK）和 p65 核转录因子-κB（p65 NF-B）的基因表达。
结果表明，YHPG 的成分，如盐酸麻黄碱、盐酸伪麻黄碱、绿原酸和大黄素，具有显著的抗病毒作用。高、中剂量 YHPG 能有效降低细胞病变效应（CPE），明显降低上清液中 IFN-和 IL-6 水平。同时，感染的 MDCK 细胞中 TLR7、MyD88、TRAF6、JNK、p38 MAPK 和 p65 NF-B 的转录水平降低。此外，不同剂量 YHPG 组之间存在一定的剂量依赖性关系。
这些结果表明，YHPG 及其成分对 H1N1 病毒的增殖具有显著的抑制作用。其机制可能与抑制 TLR7/MyD88 信号通路的激活、关键靶基因 mRNA 表达的降低以及 IFN-和 IL-6 分泌的抑制有关。
