Fig 1: Inflammatory response in PAMs induced by PRRSV 5′UTR RNA and LPS. A, B PAMs were transfected with different doses of 5′UTR RNA (1, 2, and 4 μg/well) along with 1 μg/mL LPS. qRT-PCR and Western blot results showed that compared with PAMs in other groups, PAMs in the 4 μg 5′UTR RNA and 1 μg/mL LPS co-stimulation group produced higher levels of IL-1β (p < 0.05). C After co-stimulation, the relative expression level of IL-1β mRNA obtained was similar at the 12 and 24 h time-points. D 5′UTR RNA and LPS co-stimulation induced IL-1β expression in cells and supernatants. E, F 5′UTR RNA and LPS co-stimulation induced increased levels of IL-6, IL-8 and TNF-α mRNA and protein. Expression was normalized to that of GAPDH. Different letters (a, b, c, d, and e) on data indicate significant differences between groups (p < 0.05).
Fig 2: Proposed model illustrating the anti-IL-1β activity of matrine. 5′UTR RNA and LPS co-stimulation induces the recruitment of MyD88 to activate the NF-κB signalling pathway, which in turn initiates NF-κB-dependent pro-IL-1β, IL-6, IL-8 and TNF-α transcription and also induces the activation of the NLRP3 inflammasome. Matrine treatment downregulates MyD88, NLRP3 and caspase-1 expression, inhibits ASC speck formation, suppresses IκBα phosphorylation, and interferes with the translocation of NF-κB from the cytoplasm to the nucleus.
Fig 3: Matrine inhibited the secretion of inflammatory mediators. A Matrine inhibited the mRNA expression of IL-1β, IL-6, IL-8 and TNF-α. After 6 h of co-stimulation, different doses of matrine (0.4, 0.2 and 0.1 mg/mL) or 0.04 mg/mL dexamethasone were added to the cells for an additional 6 h of incubation. The expression levels of IL-1β, IL-6, IL-8 and TNF-α genes in PAMs from all treatment groups were measured using qRT-PCR. B, C Matrine reduced the protein levels of IL-1β, IL-6, IL-8 and TNF-α. Expression was normalized to GAPDH expression. #Indicates that the expression was significantly different compared to that of the group with 5′UTR RNA and LPS co-stimulation (p < 0.05).
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