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KMID : 0364320190440010029
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Journal of East-West Medicine
2019 Volume.44 No. 1 p.29 ~ p.42
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Chungsanggyuntong-tang inhibits lipopolysaccharide-activated inflammation in RAW264.7 cells
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Seo Ho-Seok
Jee Seon-Young
Park Jong-Hyun
Hwangbo Min
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Abstract
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Backgrounds : Chungsanggyuntong-tang (CGT) is an herbal prescription for treating headache and migraine in Traditional Korean Medicine. Present study investigated in vitro anti-inflammatory activity of CGT extract in lipopolysaccharide (LPS)-activated RAW264.7 cells and elucidated signaling molecules involved.
Methods : To examine the anti-inflammatory activity of CGT, RAW264.7 cells were pretreated with 0.01-0.3 ㎎/㎖ of CGT for 1 h, and then exposed to 1 ㎍/㎖ of LPS for indicated times. After treatment, the viability of cells was detected by adding MTT reagent. Productions of nitric oxide and proinflammatory mediators were measured in culture media. Protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were determined by immunoblot analysis. Nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs) phosphorylation and inhibitory-κBα (I-κBα) expression were investigated to explore signaling molecules involved.
Results : CGT treatment up to 0.3 ㎎/㎖ had no cytotoxicity in RAW264.7 cells. Nitric oxide production by LPS was significantly and concentration-dependently inhibited by pretreatment with 0.1-0.3 ㎎/㎖ of CGT. Release of prostaglandin E2, interleukin-1β, interleukin-6, tumor necrosis factor-α, and monocyte chemoattractant protein-1 by LPS were significantly decreased by GCT pretreatment (0.1-0.3 ㎎/㎖). In addition, CGT attenuated LPS-mediated iNOS and COX-2 expression. Moreover, CGT significantly inhibited I-κBα expression and NF-κB phosphorylation, while CGT did not change the phosphorylation level of MAPKs by LPS.
Conclusions : CGT would be useful herbal prescription to reduce acute inflammation of macrophage, which mediated by inhibition of NF-κB.
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KEYWORD
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Acute inflammation, Chungsanggyuntong-tang (CGT), Lipopolysaccharide (LPS), Nuclear factor-κB (NF-κB), RAW264.7 cells
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