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Curcumin pyrazole blocks lipopolysaccharide-induced inflammation via suppression of JNK activation in RAW 264.7 macrophages.
Asian Pacific Journal of Allergy and Immunology 2018 September
BACKGROUND: Targeting inflammatory macrophages and their products is an effective method for controlling inflammation. The pyrazole analog of curcumin (curcumin pyrazole, PYR) has been reported to possess superior anti-inflammatory activity to curcumin (CUR). However, the role of PYR anti-inflammatory activity in macrophages has not yet been elucidated.
OBJECTIVE: To examine the anti-inflammatory effects of PYR and CUR in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages and determine the role of mitogen-activated protein kinases (MAPK) in their activity.
METHODS: Nitrite level was investigated by the Griess assay. The expression of inducible nitric oxide (NO) synthase, cyclooxygenase-2 (COX-2), and MAPK proteins were analyzed by western blot analysis. The pro-inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay.
RESULTS: LPS-induced NO secretion in RAW 264.7 macrophages was potently inhibited by PYR (IC50 = 3.7 ± 0.16 μM), at a higher efficacy than CUR (IC50 = 11.0 ± 0.59 μM). Treatment with identical concentrations of PYR and CUR demonstrated that PYR drastically inhibited iNOS and COX-2 expression, whereas CUR only blocked COX-2. PYR reduced the LPS-induced secretion of TNF-α to a greater extent than CUR and both similarly reduced IL-1β and IL-6 levels. Activation of c-Jun N-terminal kinase (JNK) MAPK was significantly decreased in LPS-activated RAW 264.7 macrophages upon PYR but not CUR treatment.
CONCLUSION: PYR exhibited a more potent anti-inflammatory activity than CUR. This activity is partly mediated by PYR-depended inhibition of the JNK signaling pathway and underscores the utility of PYR as an anti-inflammatory agent in macrophages.
OBJECTIVE: To examine the anti-inflammatory effects of PYR and CUR in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages and determine the role of mitogen-activated protein kinases (MAPK) in their activity.
METHODS: Nitrite level was investigated by the Griess assay. The expression of inducible nitric oxide (NO) synthase, cyclooxygenase-2 (COX-2), and MAPK proteins were analyzed by western blot analysis. The pro-inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay.
RESULTS: LPS-induced NO secretion in RAW 264.7 macrophages was potently inhibited by PYR (IC50 = 3.7 ± 0.16 μM), at a higher efficacy than CUR (IC50 = 11.0 ± 0.59 μM). Treatment with identical concentrations of PYR and CUR demonstrated that PYR drastically inhibited iNOS and COX-2 expression, whereas CUR only blocked COX-2. PYR reduced the LPS-induced secretion of TNF-α to a greater extent than CUR and both similarly reduced IL-1β and IL-6 levels. Activation of c-Jun N-terminal kinase (JNK) MAPK was significantly decreased in LPS-activated RAW 264.7 macrophages upon PYR but not CUR treatment.
CONCLUSION: PYR exhibited a more potent anti-inflammatory activity than CUR. This activity is partly mediated by PYR-depended inhibition of the JNK signaling pathway and underscores the utility of PYR as an anti-inflammatory agent in macrophages.
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