TMT-Based Quantitative Proteomic Analysis Reveals Downregulation of ITGAL and Syk by the Effects of Cycloastragenol in OVA-Induced Asthmatic Mice

Abstract

Background. Cycloastragenol (CAG) has been reported to alleviate airway inflammation in ovalbumin- (OVA-) induced asthmatic mice. However, its specific mechanisms remain unclear. Objective. This study is aimed at investigating the effects of CAG on asthma, comparing its efficacy with dexamethasone (DEX), and elucidating the mechanism of CAG’s regulation. Methods. The asthma mouse model was induced by OVA. CAG at the optimal dose of 125 mg/kg was given every day from day 0 for 20-day prevention or from day 14 for a 7-day treatment. We observed the preventive and therapeutic effects of CAG in asthmatic mice by evaluating the airway inflammation, AHR, and mucus secretion. Lung proteins were used for TMT-based quantitative proteomic analysis to enunciate its regulatory mechanisms. Results. The early administration of 125 mg/kg CAG before asthma happened prevented asthmatic mice from AHR, airway inflammation, and mucus hypersecretion, returning to nearly the original baseline. Alternatively, the administration of CAG during asthma also had the same therapeutic effects as DEX. The proteomic analysis revealed that the therapeutical effects of CAG were associated with 248 differentially expressed proteins and 3 enriched KEGG pathways. We then focused on 3 differentially expressed proteins (ITGAL, Syk, and Vav1) and demonstrated that CAG treatment downregulated ITGAL, Syk, and Vav1 by quantitative real-time PCR, western blot analysis, and immunohistochemical staining. Conclusion. These findings suggest that CAG exerts preventive and protective effects on asthma by inhibiting ITGAL, Syk, and the downstream target Vav1.

Icariin attenuates asthmatic airway inflammation via modulating alveolar macrophage activation based on network pharmacology and in vivo experiments

“…SYK is a kinase that regulates the signal transformation of adaptive immune receptors, increasing ligand affinity for integrin receptors. 27 Prior research has noted the importance of Syk because it not only acts as an inflammatory mediator, but also participates in airway wall remodeling in asthma. In addition, Syk inhibition diminishes pulmonary airway inflammation and M1 polarization.…”

“…After tracheostomized, mice were intubated and connected to the Fine Pointe Resistance and Compliance system (DSI Buxco Electronics, Saint Paul, MN, USA). To detect lung function, mice were exposed to Mch at increasing doses of 0, 6.25, 12.5 and 25 mg/ml after stable breathing 27 . Airway resistance ( R L ) and lung dynamic compliance (Cdyn) were obtained and analyzed by FinePointe data acquisition and analysis software (DSI Buxco Electronics).…”

“…To detect lung function, mice were exposed to Mch at increasing doses of 0, 6.25, 12.5 and 25 mg/ml after stable breathing. 27 Airway resistance (R L ) and lung dynamic compliance (Cdyn) were obtained and analyzed by Fine-Pointe data acquisition and analysis software (DSI Buxco Electronics).…”

“…[23][24][25] Thus, plant-derived natural products, which derive from edible herbs, have received increased attention. 26,27 Icariin (ICA) is a flavonoid glucoside derived from Epimedium, an herbal medicine and dietary supplement that has been used to treat clinical respiratory diseases. [28][29][30][31][32] Our previous clinical study has demonstrated that ICA-rich Chinese medicine prescriptions can significantly alleviate airway inflammation in patients with asthma.…”

Effects of increasing sensitizing doses of ovalbumin on airway hyperresponsiveness in asthmatic mice

“…[7][8][9][10] The previous study has demonstrated that intraperitoneal injection of OVA and alum once a week for 2 weeks is the most straightforward and effective approach to induce an asthmatic reaction in mice. 11 However, the inconsistent use of sensitizing OVA doses in current studies [12][13][14] and the lack of research on their effect on AHR necessitate further investigation. Therefore, we conducted a study using increasing doses (0, 10, 20, 50, and 100 μg) of sensitizing OVA in a BALB/c murine model of asthma to evaluate their impact on AHR to methacholine (MCh), as well as histological changes, eosinophilic infiltration, and epithelial damage.…”