目的 通过脂多糖(LPS)诱导活化小鼠单核巨噬细胞系RAW264.7建立炎症模型,探讨小钻木脂素类化合物戈米辛R对炎症细胞增殖及炎症因子分泌的影响,评价其抗炎活性及机制。方法 采用MTT比色法检测经LPS诱导的RAW264.7细胞存活率并计算半数抑制浓度(IC50);采用ELISA法检测细胞上清液TNF-α、IL-1β、IL-6等炎症因子含量;利用RT-PCR法检测细胞TNF-α、IL-1β、IL-6、IκB-α、NF-κB p65的mRNA表达量;Western blot法检测细胞IκB-α及NF-κB p65蛋白表达量。结果 与LPS模型组比较,戈米辛R可显著抑制LPS诱导的RAW264.7细胞增殖;降低细胞分泌TNF-α、IL-1β及IL-6的含量;升高LPS所致降低的IκB-α mRNA的表达量同时降低LPS引起升高的TNF-α、IL-1β、IL-6、NF-κB p65的mRNA表达量;升高IκB-α并降低NF-κB p65的蛋白表达量,显示出良好的剂量依赖性,具有统计学差异(P<0.05)。结论 戈米辛R可通过抑制TNF-α、IL-1β、IL-6、NF-κB p65的mRNA和NF-κB p65蛋白的表达,增加IκB-α mRNA和IκB-α蛋白的表达,减少致炎因子TNF-α、IL-1β、IL-6的释放,从而发挥显著抗炎活性。
Abstract
OBJECTIVE To explore the effects of gomisin R(the lignans from Kadsura longipedunculata Finet et Gagnep.) on inflammatory cell proliferation and inflammatory factor secretion, and evaluate its anti-inflammatory activity and mechanism.METHODS Inflammation model was built by inducing and activating RAW264.7, the macrophage-monocyte lineage of rats, with lipopolysaccharide(LPS), then the cell survival rate of RAW264.7 was examined via MTT, and the half inhibition concentration(IC50) was calculated. The levels of inflammatory factors, such as TNF-α, IL-1β, IL-6 in cell supernatant were tested via ELISA. The mRNA expression levels of cellular TNF-α, IL-1β, IL-6, IκB-α and NF-κB p65, were measured via RT-PCR. And the protein expression levels of cellular IκB-α and NF-κB p65, were measured via Western blot.RESULTS Compared with the LPS model set, gomisin R could notably inhibit the cell proliferation of RAW264.7 induced by LPS, decrease the TNF-α, IL-1β and IL-6 secretion of cells, enhance the mRNA expression levels of IκB-α, which were decreased by LPS, and in the meantime, lower the mRNA expressions of TNF-α, IL-1β, IL-6 and NF-κB p65, which was increased by LPS, and increase the protein expression of IκB-α while reduce that of NF-κB p65, the result of which was highly dependent on dose and the difference was statistically significant(P<0.05).CONCLUSION Gomisin R can inhibit the mRNA and NF-κB p65 protein expressions of TNF-α, IL-1β, IL-6 and NF-κB p65, and meanwhile increase the protein expressions of IκB-α mRNA and IκB-α, thus reducing the release of inflammatory factors such as TNF-α, IL-1β and IL-6, and bringing remarkable anti-inflammatory activity into full play.
关键词
戈米辛R /
抗炎 /
RAW264.7 /
细胞因子 /
NF-κB
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Key words
gomisin R /
anti-inflammatory /
RAW264.7 /
cell factor /
NF-κB
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中图分类号：
R965
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脚注
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基金
国家自然科学基金项目资助(81660721);2017年广西科技重大专项资助(桂科AA18242040);2020年广西高校中青年教师科研基础能力提升项目资助(2020KY43007)
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