Combining Network Pharmacology and Metabolomics to Reveal the Mechanism of Liu Shen Capsules Against Inflammatory and Influenza H3N2 Virus

Abstract

Ethnopharmacological relevance: Liu Shen capsule (LS) is a traditional Chinese medicine (TCM) with certain medicinal value. It has various pharmacological effects include: anti-inflammatory and antiviral activities. However, current research on the H3N2 virus and the mechanism is limited.Aim of the studyIn this research, we employed metabolomics and network pharmacology to identify the effective targets and signal pathways of LS, confirmed its activities of anti-viral and anti-inflammatory induced by H3N2 of LS and explore its probable molecular processes.

Methods: By identifying cytopathic effects (CPE) in Madin-Darby canine kidney cell (MDCK) cells, the antiviral properties of LS were assessed. Mice were used to study morbidity, lung index, viral titer, and pathological alterations. Metabolomics and network pharmacology were used to conduct the initial investigation into the antiviral mechanism of LS. The pertinent metabolite pathways were investigated, and network analysis was used to further uncover the potential targets of LS for the therapy of H3N2. With the help of immunohistochemistry and western blotting, the NF-B pathway was further confirmed.

Results: The findings demonstrated that LS inhibited progeny viral proliferation following LS treatment for 0, 1, 2, and 4 hours in vitro. Moreover, 80 and 50 mg/kg of LS significantly increased mouse survival time, decreased virus titer and lung index, and improved lung tissue lesions. Moreover, LS can drastically lower the expression of the IL-1β, TNF-α, IL-6, and NP proteins. 33 signaling pathways and 27 distinct metabolites relevant to LS were investigated using a metabolomic method. 320 potential targets and 118 active components were identified by the network pharmacology analysis. In addition, 10 important targets, 3 important metabolic pathways, and 5 important pathways linked with LS against the influenza A virus were examined by metabolomics and network pharmacology. By preventing the NF-κB pathway from being activated, LS and the NF-κB inhibitor BAY11-7082 both lowered the levels of NP and inflammatory cytokines.

Conclusions: In conclusion, this study demonstrated that LS controlled the biological network and metabolic state by a combination of network pharmacology and metabolomics in multi-target and multi-pathway systems. LS can reduce lung damage brought on by H3N2 and decrease viral growth by altering important NF-κB pathway targets.

Note:
Funding Information: This work was supported by National Key R&D Program of China (2021YFC1712904), National Natural Science Foundation of China (82004033, 82174053, 81903957), Natural Science Foundation of Guangdong Province (2020A1515111155), Macao Science and Technology Development Fund (0172/2019/A3), Guangdong Provincial Department of Natural Resources project (GDNRC [2021]51), Guangzhou Science and Technology Bureau project (202102100003, 202201020449), Traditional Chinese Medicine Bureau of Guangdong Province (20211178), The Open Project of State Key Laboratory of Component Traditional Chinese Medicine (CBCM2020101, CBCM2021102), General funded scientific research project of Hunan Provincial Health Commission (B202319018491), Guangdong Basic and Applied Basic Research Foundation(2020B1515120045, 2020A1515110151).

Conflict of Interests: The authors declared that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Ethical Approval: The Ethics Committee of Guangzhou Medical University approved the care and use of the animals in this research (2019-645).

Keywords: Liu Shen capsule, metabolomics, Network pharmacology, influenza A/Aichi/2/68 (H3N2) virus, NF-кB