Liu Shen Wan regulates the SPHK1/S1P axis to ameliorate influenza-induced inflammation via integrated network pharmacology and lipidomics.
Liu Shen Wan (LSW) can modulate sphingolipid metabolism, which is a key pathway in inflammatory regulation, yet the precise mechanistic actions remain elusive. This study aimed to elucidate the mechanism by which LSW regulates sphingolipid metabolism to mitigate influenza-induced inflammatory responses.
The potential mechanisms of LSW were initially predicted and validated via network pharmacology and lipidomics. A549 cells were infected with influenza A/Puerto Rico/8/34 (H1N1) (PR8) or transfected to overexpress sphingosine kinase-1 (SPHK1), then treated with LSW. In vivo, mice were infected with PR8 or challenged with rAAV9-SPHK1 and administered LSW for 5 days. Inflammatory factors and sphingolipid pathway-associated proteins were evaluated.
Network pharmacology identified sphingolipid signaling as a primary target of LSW. Lipidomics revealed LSW significantly reduced the levels of sphingomyelin (SM), ceramide, CerG2GNAc1, CerG3GNAc1, Ceramide phosphate and GM1 in lungs. In PR8-infected A549 cells, LSW significantly reduced sphingomyelinase (ASMase) and Ceramide (Cer) secretion. It also inhibited the expression of SPHK1 and sphingosine-1-phosphate (S1P) in A549 cells and in mice. Pharmacological inhibition of SPHK1 mirrored these anti-inflammatory effects. In SPHK1-overexpressing or TNF-α-stimulated A549 cells, LSW significantly attenuated the expression of SPHK1, CXCL10, and MCP-1. In the rAAV9-SPHK1 overexpression mouse model, LSW ameliorated lung pathological changes and reduced the expression of SPHK1, IFN-γ, and TNF-α.
LSW alleviates influenza virus-induced inflammation by inhibiting the overactivation of the sphingolipid signaling pathway, specifically through targeting the SPHK1-S1P axis and ceramide-derived lipid mediators.
The potential mechanisms of LSW were initially predicted and validated via network pharmacology and lipidomics. A549 cells were infected with influenza A/Puerto Rico/8/34 (H1N1) (PR8) or transfected to overexpress sphingosine kinase-1 (SPHK1), then treated with LSW. In vivo, mice were infected with PR8 or challenged with rAAV9-SPHK1 and administered LSW for 5 days. Inflammatory factors and sphingolipid pathway-associated proteins were evaluated.
Network pharmacology identified sphingolipid signaling as a primary target of LSW. Lipidomics revealed LSW significantly reduced the levels of sphingomyelin (SM), ceramide, CerG2GNAc1, CerG3GNAc1, Ceramide phosphate and GM1 in lungs. In PR8-infected A549 cells, LSW significantly reduced sphingomyelinase (ASMase) and Ceramide (Cer) secretion. It also inhibited the expression of SPHK1 and sphingosine-1-phosphate (S1P) in A549 cells and in mice. Pharmacological inhibition of SPHK1 mirrored these anti-inflammatory effects. In SPHK1-overexpressing or TNF-α-stimulated A549 cells, LSW significantly attenuated the expression of SPHK1, CXCL10, and MCP-1. In the rAAV9-SPHK1 overexpression mouse model, LSW ameliorated lung pathological changes and reduced the expression of SPHK1, IFN-γ, and TNF-α.
LSW alleviates influenza virus-induced inflammation by inhibiting the overactivation of the sphingolipid signaling pathway, specifically through targeting the SPHK1-S1P axis and ceramide-derived lipid mediators.
Authors
Lei Lei, Liu Liu, Xie Xie, Li Li, Cui Cui, Chen Chen, Liu Liu, Chen Chen, Li Li, Liang Liang, Liang Liang, Li Li, Zheng Zheng, Yang Yang, Ma Ma
View on Pubmed