Objective:To evaluate the effects of Capsosiphon fulvescens(C.fulvescens)ethanolic extract on inflammation in lipopolysaccharide(LPS)-induced RAW296.7 macrophages.Methods:The protective effects of C.fulvescens ethanol...Objective:To evaluate the effects of Capsosiphon fulvescens(C.fulvescens)ethanolic extract on inflammation in lipopolysaccharide(LPS)-induced RAW296.7 macrophages.Methods:The protective effects of C.fulvescens ethanolic extract on LPS-induced inflammation in RAW264.7 macrophages were assessed using biochemical analysis,including enzyme-linked immunosorbent assay,quantitative reverse transcription-polymerase chain reaction,and Western blot analysis.To examine reactive oxygen species(ROS)production,flow cytometry analysis,and immunofluorescence staining were used.Furthermore,the modulatory effect of C.fulvescens ethanolic extract on NF-κB activation was investigated.Results:C.fulvescens ethanolic extract significantly attenuated LPS-induced levels of pro-inflammatory cytokines and notably reduced the secretion and mRNA levels of LPS-mediated matrix metalloproteinases.In addition,C.fulvescens ethanolic extract decreased ROS production and suppressed the TLR4/NF-κB signaling pathway.Conclusions:C.fulvescens ethanolic extract alleviates inflammation as well as oxidative stress by modulating the TLR4/NF-κB signaling in LPS-induced RAW264.7 macrophages.C.fulvescens can be used as a potential therapeutic agent to suppress inflammation and oxidative stress-associated diseases.展开更多
Objective:To study the protective effect of paeonol on LPS-induced inflammatory injury of macrophage RAW264.7 cells and its mechanism.Methods:Macrophage RAW264.7 cells were cultured and divided into blank group,LPS(1...Objective:To study the protective effect of paeonol on LPS-induced inflammatory injury of macrophage RAW264.7 cells and its mechanism.Methods:Macrophage RAW264.7 cells were cultured and divided into blank group,LPS(1μg/mL)group,paeonol(240μmol/mL)group and TAK242(10μmol/mL)group.The cell activity was detected by CCK8 method,the cell morphology was observed by inverted microscope,the contents of GSH and MDA in cell culture medium were determined by colorimetry,the mitochondrial membrane potential was detected by JC-1 method,the expression distribution of F4/80 and p-NF-κB protein was detected by immunofluorescence method,and the expression of TLR4/MAPK/NF-κB related pathway protein was detected by Western blotting.Results:Compared with the blank group,the cell viability induced by 1μg/mL LPS was 0.4972±0.061(P<0.01),which was close to the half inhibition rate.Compared with LPS group,the expression of p-NF-κB protein in 240μmol/mL paeonol pretreated cell group was down-regulated most significantly(P<0.01),and the expression of TLR4 protein was inhibited most significantly in 10μmol/mL TAK242 pretreated cell group.Compared with LPS group(P<0.01),the cell morphology of paeonol group recovered.Decrease MDA content and increase GSH content in cell culture medium(P<0.01),In the results of mitochondrial membrane potential,the red light of paeonol group was significantly enhanced and the green light was significantly weakened(P<0.001).The expression distribution of F4/80 and p-NF-κB protein in paeonol group decreased significantly(P<0.01),and the expressions of TLR4,p-IκB,p-p38,p-JNK and p-NF-κB protein were down-regulated(P<0.05).Conclusion:Paeonol can improve the inflammatory injury of RAW264.7 cells induced by LPS,and its mechanism may be related to TLR4/MAPK/NF-κB pathway.展开更多
多种真菌多糖近年来因其免疫调节活性成为保健食品领域研究的热点。本实验以食药用真菌蛹虫草提取物蛹虫草多糖(Cordyceps militaris polysaccharides,CMP)为研究材料,初步探究CMP对小鼠巨噬细胞RAW264.7免疫活性的调节机制。噻唑蓝实...多种真菌多糖近年来因其免疫调节活性成为保健食品领域研究的热点。本实验以食药用真菌蛹虫草提取物蛹虫草多糖(Cordyceps militaris polysaccharides,CMP)为研究材料,初步探究CMP对小鼠巨噬细胞RAW264.7免疫活性的调节机制。噻唑蓝实验结果显示CMP无细胞毒性,且100、200μg/mL CMP可以明显增强RAW264.7细胞活性;中性红法、Griess法和酶联免疫吸附检测结果表明25～200μg/mL的CMP以剂量依赖方式增强RAW264.7细胞吞噬活性并增加一氧化氮(nitric oxide,NO)和白介素-1β(interleukin-1β,IL-1β)分泌,肿瘤坏死因子-α(tumor necrosisfactor-α,TNF-α)的分泌呈先升高后降低的趋势,在100μg/mL时达到最高值;抑制剂中和实验结果显示当Toll样受体4(Toll-like receptors 4,TLR4)和甘露糖受体(mannose receptor,MR)受到抑制时,CMP诱导的RAW264.7免疫反应显著降低,这表明TLR4和MR均为CMP激活巨噬细胞的受体;此外,Western blot结果显示丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)信号通路也参与CMP诱导的RAW264.7细胞分泌NO、TNF-α和IL-1β。表明TLR4和MR/MAPK信号传导途径在CMP诱导巨噬细胞RAW264.7产生免疫应答时发挥了重要作用。展开更多
基金funded by Korea Institute of Marine Science&Technology Promotion(KIMST)funded by the Ministry of Oceans and Fisheries,Korea(20220488).
文摘Objective:To evaluate the effects of Capsosiphon fulvescens(C.fulvescens)ethanolic extract on inflammation in lipopolysaccharide(LPS)-induced RAW296.7 macrophages.Methods:The protective effects of C.fulvescens ethanolic extract on LPS-induced inflammation in RAW264.7 macrophages were assessed using biochemical analysis,including enzyme-linked immunosorbent assay,quantitative reverse transcription-polymerase chain reaction,and Western blot analysis.To examine reactive oxygen species(ROS)production,flow cytometry analysis,and immunofluorescence staining were used.Furthermore,the modulatory effect of C.fulvescens ethanolic extract on NF-κB activation was investigated.Results:C.fulvescens ethanolic extract significantly attenuated LPS-induced levels of pro-inflammatory cytokines and notably reduced the secretion and mRNA levels of LPS-mediated matrix metalloproteinases.In addition,C.fulvescens ethanolic extract decreased ROS production and suppressed the TLR4/NF-κB signaling pathway.Conclusions:C.fulvescens ethanolic extract alleviates inflammation as well as oxidative stress by modulating the TLR4/NF-κB signaling in LPS-induced RAW264.7 macrophages.C.fulvescens can be used as a potential therapeutic agent to suppress inflammation and oxidative stress-associated diseases.
文摘Objective:To study the protective effect of paeonol on LPS-induced inflammatory injury of macrophage RAW264.7 cells and its mechanism.Methods:Macrophage RAW264.7 cells were cultured and divided into blank group,LPS(1μg/mL)group,paeonol(240μmol/mL)group and TAK242(10μmol/mL)group.The cell activity was detected by CCK8 method,the cell morphology was observed by inverted microscope,the contents of GSH and MDA in cell culture medium were determined by colorimetry,the mitochondrial membrane potential was detected by JC-1 method,the expression distribution of F4/80 and p-NF-κB protein was detected by immunofluorescence method,and the expression of TLR4/MAPK/NF-κB related pathway protein was detected by Western blotting.Results:Compared with the blank group,the cell viability induced by 1μg/mL LPS was 0.4972±0.061(P<0.01),which was close to the half inhibition rate.Compared with LPS group,the expression of p-NF-κB protein in 240μmol/mL paeonol pretreated cell group was down-regulated most significantly(P<0.01),and the expression of TLR4 protein was inhibited most significantly in 10μmol/mL TAK242 pretreated cell group.Compared with LPS group(P<0.01),the cell morphology of paeonol group recovered.Decrease MDA content and increase GSH content in cell culture medium(P<0.01),In the results of mitochondrial membrane potential,the red light of paeonol group was significantly enhanced and the green light was significantly weakened(P<0.001).The expression distribution of F4/80 and p-NF-κB protein in paeonol group decreased significantly(P<0.01),and the expressions of TLR4,p-IκB,p-p38,p-JNK and p-NF-κB protein were down-regulated(P<0.05).Conclusion:Paeonol can improve the inflammatory injury of RAW264.7 cells induced by LPS,and its mechanism may be related to TLR4/MAPK/NF-κB pathway.
