Anti-inflammatory effect of a sesquiterpene dimer compound(+)-chlorahupetenes B from Chloranthus henryi var.hupehensis(Pamp.)K.F.Wu on lipopolysaccharides-stimulated RAW264.7 Cells
Objective To evaluate the anti-inflammatory activity and mechanism of(+)-chlorahupetenes B,a natural eudesmane-type sesquiterpenoid dimer compound obtained from Chloranthus henryi var.hupehensis,in lipopolysaccharides(LPS)-induced RAW264.7 cell.Methods RAW264.7 cells were divided into control group(given equal volume of DMSO),model group(given equal volume of DMSO),dexamethasone sodium phosphate injection(Dex,positive control,1 μmol·L-1)group,and(+)-CHB low,medium,and high concentrations(5,10,20 μmol·L-1)groups.Each group was incubated with corresponding drugs for 1 h.Except for the control group, the other groups were induced with LPS (1 μg·mL-1) for 24 hours to create an inflammatory response model.Cell proliferation detection method was used to evaluate cell viability. Griess reaction was employed for determining nitric oxide(NO) concentration. Enzyme-linked immunosorbent assay was utilized for detecting inflammatory cytokines including tumor necrosis factor α (TNF- α), interleukin (IL)-1β, and IL-6. Real time fluorescence quantitative PCR (qRT PCR) was performed to analyze IL-1β, NOD like receptor heat protein domain associated protein 3 (NLRP3), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), IL-6, and TNF-α mRNA expression. Western blotting detection was used to detect the expression levels of Toll like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), nuclear factor κB (NF-κB) p65, p-NF-κB p65, NLRP3, and purinergic receptor (P2X7) proteins. Results Compared with the model group, (+)-CHB reduced the number of spindle cells and restored most of the cells to their normal morphology, significantly inhibited NO and TNF-α, IL-6, IL-1β generation (P ＜ 0.01),significantly reduced COX-2, iNOS, IL-6, TNF-α, NLRP3, and IL-1β mRNA levels (P ＜ 0.05, 0.01), significantly reduced the expression levels of TLR4, MyD88, NF-κB p65, p-NF-κB p65, NLRP3, and P2X7 proteins (P ＜ 0.05, 0.01). Conclusion These findings suggested that ( + ) -CHB was an anti-inflammatory compound which could relieve LPS-stimulated macrophage inflammatory response partly by targeting TLR4/MyD88/NF-κB signal pathway and P2X7/NLRP3/IL-1β inflammasome axis.
Chloranthus henryi var. hupehensis (Pamp.) K. F. Wusesquiterpene dimer compound(+)-chlorahupetenes Banti-inflammatorymacrophagesToll like receptor 4 (TLR4)nuclear factor κB (NF- κB)purinergic receptors (P2X7)NOD like receptor heat protein domain as
胡蕴然、庄鹏宇、柳航、钱铭、杨慧、葛卫红
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中国药科大学基础医学与临床药学学院,南京鼓楼医院药学部,江苏南京 210008
南京大学医学院附属鼓楼医院药学部,江苏南京 210008
华北理工大学药学院,河北唐山 063210
金粟兰 倍半萜二聚体化合物 (+)-Chlorahupetenes B 抗炎 巨噬细胞 Toll样受体4(TLR4) 核因子κB(NF-κB) 嘌呤能受体(P2X7) NOD样受体热蛋白结构域相关蛋白3(NLRP3)
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