Inhibitive Effect of Proanthocyanidins on Cyclooxygenase-2 Expression in A549 Cells Induced by Cytok

来源 :Journal of Shanghai Jiaotong University(Science) | 被引量 : 0次 | 上传用户:wangnayangyang
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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. 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 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 subunits p65 (NF-κB / p65) and the inhibitor-κB (I-κB) were measured by the Western blotting and immunocytochemistry. The results showed 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 prot ein and the degradation of I-κB protein.
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