Abstract
Ageratina adenophora is an invasive plant known for its toxicity to livestock. Current research on this plant has shifted from toxicity prevention to the beneficial utilization of plant resources. This study was performed to investigate the effects and mechanisms of cryptochlorogenic acid (CCGA) isolated from Ageratina adenophora on the inflammatory responses induced by lipopolysaccharide (LPS) in RAW264.7 cells. RAW264.7 cells were pretreated with CCGA (200, 100, and 50 μg/mL) and subsequently stimulated with LPS (1 μg/mL) for 16 h. The cytotoxicity of CCGA was tested using the Cell Counting Kit (CCK8). The mechanism of action of CCGA in attenuating inflammation was also identified using enzyme-linked immunosorbent assay (ELISA), quantitative reverse transcription-polymerase chain reaction, and Western blot. The results showed that CCGA had a maximal safe concentration of 200 mg/mL. Moreover, CCGA reduced the level of nitric oxide (NO) and iNOS in LPS-induced RAW264.7 cells (p < 0.01). In addition, CCGA reduced the levels of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-8) and cyclooxygenase-2 (COX-2) in LPS-induced RAW264.7 cells at both the mRNA and protein levels (p < 0.01). CCGA prevented the activation of nuclear factor-kappa B (NF-kB) in LPS-induced RAW264.7 cells via the inhibition of IKK and IκB phosphorylation and the degradation of IκB proteins (p < 0.01). This finding indicated that CCGA isolated from A. adenophora may be a potential candidate for the treatment of inflammation-related diseases.
Highlights
Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, These authors contributed to this work
The imported test kit for fetal bovine serum (GIBCO), the nitric oxide (NO) box, and enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-1 β, IL-6, IL-8, IL-10, and COX-2 were purchased from Nanjing Jiancheng Company Ltd., Nanjing, China
It was preliminarily inferred that the extracted monomers werecryptochlorogenic cryptochlorogenic acid, eluent the retention same retention factors (RFs) monomers were acid, andand eluent with with the same factors (RFs)
Summary
Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, These authors contributed to this work. Ageratina adenophora is an invasive plant known for its toxicity to livestock. Current research on this plant has shifted from toxicity prevention to the beneficial utilization of plant resources. This study was performed to investigate the effects and mechanisms of cryptochlorogenic acid (CCGA). Isolated from Ageratina adenophora on the inflammatory responses induced by lipopolysaccharide (LPS) in RAW264.7 cells. RAW264.7 cells were pretreated with CCGA (200, 100, and 50 μg/mL). The cytotoxicity of CCGA was tested using the Cell Counting Kit (CCK8). CCGA reduced the level of nitric oxide (NO). INOS in LPS-induced RAW264.7 cells (p < 0.01). CCGA reduced the levels of proinflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-8) and cyclooxygenase-2 (COX-2) in LPS-induced
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