摘要
目的探讨刺槐素(Acacetin)对肿瘤坏死因子-α(TNF-α)诱导的人气道平滑肌细胞(HASMCs)分泌、表达单核细胞趋化蛋白-1(monocyte chemotactic protein1,MCP-1)的影响及可能的机制。方法 CCK-8法测HASMCs增殖活性;酶联免疫吸附法(ELISA)测HASMCs上清液中MCP-1含量;实时荧光定量PCR(RT-PCR)检测MCP-1mRNA表达量;蛋白印迹法(Western blot)检测HASMCs中磷酸化P65(P-P65)蛋白的表达情况。结果①Acacetin及NF-κB抑制剂JSH-23均能显著降低TNF-α诱导的HASMCsMCP-1生成,且Acacetin的抑制作用具有浓度依赖性(P<0.05);②同时,Acacetin及JSH-23均能显著减少TNF-α诱导的MCP-1mRNA的表达(P<0.05);③Acacetin能显著降低TNF-α引起的HASMCsP65蛋白磷酸化(P<0.05)。结论降低NF-κB通路的活化水平可能是Acacetin抑制TNF-α诱导HASMCs合成、分泌MCP-1及其mRNA,从而发挥抗炎作用的机制之一。
Objective To investigate whether Acacetin can regulate TNF-α induced monocyte chemotactic protein1 (MCP-1) expression on human smooth muscle cells and its mechanism involved. Methods CCK-8 assay measured HASMCs proliferation; ELISA detected the levels of MCP-1 in the supernatants of HASMCs; Reverse transcriptase polymerase chain reaction (RT-PCR) detected the expression of MCP-1 mRNA in HASMCs; Western Blotting detectedP65 phosphorylation of HASMCs after TNF-α and/or Acacetin treatment.Results Our data showed that Acacetin and NF-κB activation inhibitor (JSH-23) could signiflcantly suppress TNF-α-induced MCP-1 protein and mRNA expression; Acacetin markedly suppressed TNF-α-induced P65 phosphorylation.Conclusions These results support that Acacetin exerts its anti-inflammatory activity by suppressing TNF-α-induced expression of MCP-1 via blockage of activation of NF-κB.
出处
《中华哮喘杂志（电子版）》
CAS
2013年第3期6-9,共4页
Chinese Journal of Asthma(Electronic Version)
基金
科技部科技重大专项"重大新药创制"
基金编号:2011zx09302-003-02
江苏高校优势学科建设工程资助项目
