冠突散囊菌来源的2-6-H-5-B化合物的抗炎作用及机制研究

Study on Anti-inflammatory Activity and Mechanism of Compound 2-6-H-5-B from Eurotium Cristatum

目的研究冠突散囊菌来源的苯甲醛衍生物(2-6-H-5-B)对脂多糖诱导的RAW264.7细胞的抗炎作用及机制。方法RAW264.7细胞经不同浓度的苯甲醛衍生物(2-6-H-5-B)(25,50和100μmol·L^(-1))及阳性对照药地塞米松(0.5μmol·L^(-1))预处理1 h后,除对照组外,其余各组给予100 ng·mL^(-1)脂多糖刺激诱导24 h。采用Griess试剂法检测NO释放水平,ELISA法分析TNF-α、IL-6和MCP-1的表达水平,以及Western blotting检测AMPK、NF-κB和MAPK信号通路的蛋白表达情况。结果化合物2-6-H-5-B可以显著抑制活化的巨噬细胞NO(P<0.01或P<0.05)、TNF-α(P<0.01)、IL-6(P<0.01)和MCP-1(P<0.01)的产生,表现出较好的抗炎活性。研究表明,2-6-H-5-B未能明显阻断NF-κB和MAPK信号通路的活化,但可以显著增加p-AMPK的表达。结论2-6-H-5-B是一个较有潜力的抗炎药物先导物,其抗炎活性可能与激活AMPK通路相关。

OBJECTIVE To study the effects and mechanisms of anti-inflammatory of benzaldehyde derivatives(2-6-H-5-B)derived from Eurotium cristatum in lipopolysaccharide stimulated RAW264.7 cells.METHODS RAW264.7 cells were pretreated with benzaldehyde derivative(compound 2-6-H-5-B)(25,50,100μmol·L^(-1))and postive control drug dexamethasone(0.5μmol·L^(-1))for 1 h,then cells were treated with 100 ng·mL^(-1)lipopolysaccharide for 24 h except control group.The nitrite concentration in the culture medium was measured as an indicator of NO production by the Griess reagent according to the manufacturer’s protocol.The expression levels of TNF-α,IL-6 and MCP-1 in the cultured supernatant were detected by ELISA kits,and the protein expressions of AMPK,NF-κB and MAPK signaling pathways were analyzed by Western blotting.RESULTS The expression of pro-inflammatory cytokines,including TNF-α(P<0.01),IL-6(P<0.01)and MCP-1(P<0.01),as well as the production of NO(P<0.01 or P<0.05)were inhibited by compound 2-6-H-5-B.Further mechanism studies showed that 2-6-H-5-B failed to inhibit the activation of NF-κB and MAPK signaling pathways,but increased the expression of p-AMPK significantly.CONCLUSION The 2-6-H-5-B is a potential anti-inflammatory drug,and its anti-inflammatory activity may be related to the activation of AMPK pathway.