两种细叶勾儿茶单体化合物的抗炎活性相关研究OA

Objective:To screen anti-inflammation of No 1 naphthyl-α-pyranone monomer compounds and No 2 chromenone monomer compounds separated and extracted from berchemia lineata,and to investigate its anti-inflammatory mechanism.Methods:0.5 μg/mL lipopolysaccharide(LPS)was added to mouse macrophages RAW264.7 to active the cells and trigger the inflammatory response after LPS extracellular stimulus for 12 h;CCK-8 method was applied to detect the effects of monomer compounds No 1 and 2 on the survival rates of RAW264.7 cells when the concentrations were 50 and 75 μM;when concentration gradient(0,6.25,12.5,25,50 and 75 μM)was set for monomer compounds No 1 and 2,the contents of NO contained in RAW264.7 cellular supernatant were detected by Griess method,and NO inhibition rate was calculated,so as to measure anti-inflammatory activity gradient;ELISA method was adopted to measure the influence of monomer compounds No 1 after co-incubating with LPS for 12 on the levels of TNF-α,IL-6 and NF-κB;and to measure the effects of monomer compounds No 2 after co-incubating with LPS for 12 on the levels of IL-6,TNF-α,and NF-κB,iNOS and COX-2;PCR was used to detect the influence of monomer compounds No 1 after co-incubating with LPS for 12 on the expressions of IL-1β,IL-6 and iNOS mRNA in RAW264.7 cells;the effects of monomer compounds No 2 on the expressions of TNF-α,COX-2 and iNOS mRNA in LPS-induced RAW264.7 cells were measured via PCR method.Results:Two compounds separated from the herb showed no significant impacts on the survival rate of RAW264.7 cells at the concentrations of 50 and 75 μΜ;the detection results of the contents of NO contained in cellular supernatant displayed that the concentration of NO was reduced markedly as the concentrations of the compounds increased;monomer compounds No 1 could lower the levels of IL-6,TNF-α,and NF-κB in cellular supernatant of LPS-induced RAW264.7 cells,monomer compounds No 2 could reduce the expressions of TNF-α,IL-6,and NF-κB,iNOS and COX-2.Monomer compounds No 1 could inhibit the expressions of IL-6,IL-1β and iNOS mRNA in LPS-induced RAW264.7 cells;monomer compounds No 2 could restrain the expressions of TNF-α,iNOS and COX-2 mRNA.Conclusion:Two compounds extracted from the herb have no obvious drug toxicity,and certain anti-inflammatory activity,the anti-inflammatory activity was increased evidently with the increase of compound concentration.Two compounds could exert anti-inflammatory effects possibly through regulating NF-κB signaling pathway.