Inhibitive Effect of Proanthocyanidins on Cyclooxygenase-2 Expression in A549 Cells Induced by Cytokine Interleukin-1 Beta

【摘要】 Cyclooxygenase-2(COX-2),an important enzyme,plays a pathological role in diseases,which can be inhibited by proanthocyanidins(PCs) effectively.In this paper,we investigated the inhibitive mechanism of COX-2 performed by PCs.Reverse transcription-polymerase chain reaction(RT-PCR) was performed to identify the mRNA expression level of COX-2 in A549 cell,which was induced by interleukin-1 beta(IL-1β).The pGL3 luciferase reporter vector containing the COX-2 gene promoter fragment(pGL3/COX-2p) was transfected into A549 cell induced by IL-1β,the interference on the COX-2 promoter activity from PCs was analyzed using a dualluciferase reporter assay,and the expressions of the nuclear factor κB composed of subunit p65(NF-κB/p65) and the inhibitor-κB(I-κB) were measured by the Western blotting and immunocytochemistry.The results exhibited that PCs not only inhibited the transcript of COX-2 mRNA and the COX-2 promoter activity,but also suppressed the nuclear translocation of NF-κB/p65 protein and the degradation of I-κB protein.更多还原

【Abstract】 Cyclooxygenase-2(COX-2),an important enzyme,plays a pathological role in diseases,which can be inhibited by proanthocyanidins(PCs) effectively.In this paper,we investigated the inhibitive mechanism of COX-2 performed by PCs.Reverse transcription-polymerase chain reaction(RT-PCR) was performed to identify the mRNA expression level of COX-2 in A549 cell,which was induced by interleukin-1 beta(IL-1β).The pGL3 luciferase reporter vector containing the COX-2 gene promoter fragment(pGL3/COX-2p) was transfected into A549 cell induced by IL-1β,the interference on the COX-2 promoter activity from PCs was analyzed using a dualluciferase reporter assay,and the expressions of the nuclear factor κB composed of subunit p65(NF-κB/p65) and the inhibitor-κB(I-κB) were measured by the Western blotting and immunocytochemistry.The results exhibited that PCs not only inhibited the transcript of COX-2 mRNA and the COX-2 promoter activity,but also suppressed the nuclear translocation of NF-κB/p65 protein and the degradation of I-κB protein.更多还原