Tanreqing Injection Inhibits Activation of NLRP3 Inflammasome in Macrophages Infected with Influenza A Virus by Promoting Mitophagy

Tian-Yi Liu; Yu Hao; Qin Mao; Na Zhou; Meng-Hua Liu; Jun Wu; Yi Wang; Ming-Rui Yang

doi:10.1007/s11655-024-3905-3

Tanreqing Injection Inhibits Activation of NLRP3 Inflammasome in Macrophages Infected with Influenza A Virus by Promoting Mitophagy

Chin J Integr Med. 2024 Jun 24. doi: 10.1007/s11655-024-3905-3. Online ahead of print.

Authors

Tian-Yi Liu¹, Yu Hao¹, Qin Mao¹, Na Zhou¹, Meng-Hua Liu¹, Jun Wu¹, Yi Wang², Ming-Rui Yang³

Affiliations

¹ School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 102488, China.
² Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
³ School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 102488, China. mingruiyang@126.com.

PMID: 38910190
DOI: 10.1007/s11655-024-3905-3

Abstract

Objective: To investigate the inhibitory effect of Tanreqing Injection (TRQ) on the activation of nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3 (NLRP3) inflammasome in macrophages infected with influenza A virus and the underlying mechanism based on mitophagy pathway.

Methods: The inflammatory model of murine macrophage J774A.1 induced by influenza A virus [strain A/Puerto Rico/8/1934 (H1N1), PR8] was constructed and treated by TRQ, while the mitochondria-targeted antioxidant Mito-TEMPO and autophagy specific inhibitor 3-methyladenine (3-MA) were used as controls to intensively study the anti-inflammatory mechanism of TRQ based on mitophagy-mitochondrial reactive oxygen species (mtROS)-NLRP3 inflammasome pathway. The levels of NLRP3, Caspase-1 p20, microtubule-associated protein 1 light chain 3 II (LC3II) and P62 proteins were measured by Western blot. The release of interleukin-1β (IL-1β) was tested by enzyme linked immunosorbent assay, the mtROS level was detected by flow cytometry, and the immunofluorescence and co-localization of LC3 and mitochondria were observed under confocal laser scanning microscopy.

Results: Similar to the effect of Mito-TEMPO and contrary to the results of 3-MA treatment, TRQ could significantly reduce the expressions of NLRP3, Caspase-1 p20, and autophagy adaptor P62, promote the expression of autophagy marker LC3II, enhance the mitochondrial fluorescence intensity, and inhibit the release of mtROS and IL-1β (all P<0.01). Moreover, LC3 was co-localized with mitochondria, confirming the type of mitophagy.

Conclusion: TRQ could reduce the level of mtROS by promoting mitophagy in macrophages infected with influenza A virus, thus inhibiting the activation of NLRP3 inflammasome and the release of IL-1β, and attenuating the inflammatory response.

Keywords: Chinese medicine; Tanreqing Injection; anti-inflammation; influenza virus; mitophagy; nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3 inflammasome.