Levistilide A Ameliorates NLRP3 Expression Involving the Syk-p38/JNK Pathway and Peripheral Obliterans in Rats

Figure 2

LA inhibited Syk and p38/JNK activity. HUVECs were pretreated in the presence of various doses of LA for 1 h and stimulated with LPS for 30 min. A Western blotting analysis was performed to evaluate the levels of phosphorylated Syk and total Syk (a). HUVECs were treated with 50 μM of LA for 1 h before LPS stimulation for 15 min, and then the total proteins of cells were extracted. The activation of the MAPK (b) pathways was detected using Western blotting with specific antibodies. Syk activity was analyzed using an in vitro kinase assay through recombinant Syk obtained from Promega. The assay (ADP-Glo kinase assay) was performed to measure luminescent signals by using Ultra-Glo Luciferase; its ADP from a kinase reaction is converted into ATP converted into light. Kinase assays were conducted in the presence of various doses of LA for 30 min. The results were evaluated using Syk activity (c). Data represent the of at least three independent experiments, and each experiment was performed in triplicate. and versus control alone; versus LPS alone.

(a)

(b)

(c)