Xiaqing Wu,1,2,* Xin Li,2,* Ying Chai,1,2 Yushan Tian,1,2 Hongjuan Wang,1,2 Xiao Li,1,2 Jingzheng Zhang,2 Chunmei Guang,2 Enliang Hong,2 Haoping Cheng,2 Qingyuan Hu,1,2 Huan Chen,1,2 Hongwei Hou1,2 

1Key Laboratory of Tobacco Biological Effects, China National Tobacco Quality Supervision & Test Center, Zhengzhou, Henan, People’s Republic of China; 2Beijing Life Science Academy, Beijing, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Huan Chen; Hongwei Hou, Key Laboratory of Tobacco Biological Effects, China National Tobacco Quality Supervision & Test Center, New & High-tech Industry Development Zone, No.6 Cuizhu Street, Zhengzhou, Henan, 450001, People’s Republic of China, Email hunny_ch@163.com; qsfctc@163.com

Introduction: Cordyceps sinensis, an entomogenous fungus with unique biological properties, has demonstrated significant anti-inflammatory potential. However, its effects on inflammation regulation need to be further investigated in detail.
Methods: In this study, we aimed to analyze the Cordyceps sinensis extract (CSE) obtained via ethanol extraction and to assess its effects on inflammation regulation. The secretion of pro-inflammatory cytokines (IL-6, TNF-α, IL-8, and IL-1β) and the level of MMP9, Nrf2/HO-1 and ROS were evaluated. A transwell system with THP-1 and BEAS-2B cells was used to explore the inflammatory damage. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses were conducted on the differentially expressed genes.
Results: CSE exhibited no cytotoxicity to THP-1 cells at concentrations ≤ 1.6 mg/mL. Treatment of LPS-induced THP-1 cells with CSE significantly inhibited the secretion of pro-inflammatory cytokines. CSE reduced inflammation-related protein MMP9, while upregulating the anti-inflammatory Nrf2/HO-1 signaling pathway. Fluorescence assays using DCF and JC-1 further confirmed that CSE help mitigate oxidative stress-induced inflammation. CSE treatment protected BEAS-2B cells from inflammatory damage. Moreover, the immune system process was a shared GO term between LPS-only treatment and combined LPS and CSE treatment. KEGG enrichment analysis showed that CSE is capable of regulating genes associated with inflammatory and anti-inflammatory responses.
Conclusion: These findings highlight the potential of CSE as an immune-regulating agent in functional foods and health products.

Keywords: Cordyceps sinensis, anti-inflammatory, macrophage, transcriptomic analyses