Conciliatory Anti-Allergic Decoction Attenuates Pyroptosis in RSV-Infected Asthmatic Mice and Lipopolysaccharide (LPS)-Induced 16HBE Cells by Inhibiting TLR3/NLRP3/NF-κB/IRF3 Signaling Pathway

Abstract

Respiratory syncytial virus (RSV) infection can deteriorate asthma by inducing persistent airway inflammation. Increasing evidence elucidated that pyroptosis plays a pivotal role in asthma. Conciliatory anti-allergic decoction (CAD) exhibits an anti-inflammatory effect in ovalbumin (OVA)-induced asthma; however, the effects and mechanisms of CAD in RSV-infected asthmatic mice have not yet been elucidated. The RSV-infected asthmatic mice model and lipopolysaccharide (LPS)-induced 16HBE cell pyroptosis model were established, respectively. Pulmonary function, ELISA, and histopathologic analysis were performed to assess the airway inflammation and remodeling in mice with CAD treatment. Furthermore, ultra-performance liquid chromatography-quadrupole time-of-flight/mass spectrometry (UPLC-Q-TOF/MS) was conducted to identify the chemical compounds of high-dose CAD (30 g/kg). Cell viability and apoptosis of 16HBE cells were assessed by CCK-8 and flow cytometry assays, respectively. Finally, the expression levels of apoptosis-, pyroptosis-, and TLR3/NLRP3/NF-κB/IRF3 signaling-related genes were measured with qRT-PCR or western blotting, respectively. Pulmonary function tests showed that CAD significantly ameliorated respiratory dysfunction, airway hyperresponsiveness, inflammation cell recruitment in BALF, pulmonary inflammation, collagen deposition, and cell death in lung tissues. CAD significantly decreased the content of TNF-α, IL-13, IL-4, IL-1β and IL-5 in the bronchoalveolar lavage fluid (BALF), IL-17, IL-6, and OVA-specific IgE in serum and increased serum IFN-γ in asthma mice. The results of UPLC-Q-TOF/MS showed that high-dose CAD had 88 kinds of chemical components. In vitro, CAD-contained serum significantly suppressed LPS-induced 16HBE cell apoptosis. Additionally, CAD and CAD-contained serum attenuated the up-regulated expressions of Bax, Cleaved caspase-3, NLRP3, ASC, Cleaved caspase-1, GSDMD-N, IL-18, IL-1β, TLR3, p-P65, p-IκBα, and IRF3 but increased Bcl-1 and GSDMD levels in the asthma mice and LPS-induced 16HBE cells, respectively. These results illustrated that CAD may have a potential role in improving airway inflammation and pyroptosis through inhibition of the TLR3/NLRP3/NF-κB/IRF3 signaling pathway.

Myeloid-specific knockout of Notch-1 inhibits MyD88- and TRIF-mediated TLR signaling pathways by regulating oxidative stress-SHP2 axis, thus restraining aneurysm progression

“…Then phagocytosis assay with fluorescent microspheres was designed, and it was found that myeloid-specific knockout of Notch-1 promoted the phagocytic capacity of macrophages by promoting the SHP2 signaling pathway. Other research suggests that ISL exerts its anti-inflammatory effects by inhibiting the activation of the Notch-1/NF-κB and MAPK signaling pathways, which will be the direction of our future research [ 25 – 27 ]. In conclusion, myeloid-specific knockout of Notch-1 can down-regulate the expression of TLR signaling pathway-related proteins, inhibit the oxidative stress level and increase the protein expression of SHP2, thereby relieving the development and formation of aneurysms.…”

Oxidative stress and ROS-mediated cellular events in RSV infection: potential protective roles of antioxidants

“…Many lines of evidence suggest that RSV can activate the NLRP3 inflammasome and induce inflammasome-related airway inflammation, even pyroptosis [ 70 , 76 , 77 ], similar to other RNA viruses [ 73 ]. As we mentioned before, RSV can induce excessive production of ROS [ 15 , 54 ], which contributes to the activation of the NLRP3 inflammasome in RSV infection (Fig.…”

“…Although the NLRP3 inflammasome is a key player in antiviral responses [ 65 ], it also leads to an excessive innate immune response, which is implicated in RSV-induced lung inflammation and damage. Multiple interventions and treatments have been reported to attenuate and inhibit the activation of the NLRP3 inflammasome during RSV infection, which abrogates lung inflammatory injury and long-term airway disease development [ 76 , 77 , 80 , 86 ]. Thus, it is plausible to infer that inhibiting ROS production may serve as a potential measure to prevent ROS-mediated NLRP3 inflammasome activation and lytic cell death, which may ameliorate RSV-induced lung inflammatory damage.…”

Molecular mechanisms of neuronal death in brain injury after subarachnoid hemorrhage

“…B. et al, 2019 ). In the classic Caspase-1-induced cell pyroptosis process, inflammasomes (NLRP1, NLRP3, NLRC4 and AIM2) sense the danger signal and induce the connector molecule ASC to activate Caspase-1, thus cleaving GSDMD ( Swanson et al, 2019 ; Chen et al, 2022 ). Subsequently, cellular pores are formed on the cell membrane, which results in water and ion influx and ultimately cell swelling and death ( Shi et al, 2015 ; Broz and Dixit, 2016 ).…”