目的建立环氧化酶Ⅱ活性的检测模型，提供环氧化酶Ⅱ抑制剂的筛选模型及非甾体类抗炎药(nonsteoroidal antiinflammatory drug,NSAID)筛选的方法。方法利用NIH 3T3细胞在血清存在情况下能诱导生成环氧化酶Ⅱ的特性，通过加入底物花生四烯酸，检测终产物前列腺素E-2(PGE-2)含量的变化，判断化合物对环氧化酶Ⅱ的作用。以rofecoxib(罗非昔布)为阳性药来判断该模型能否用于环氧化酶Ⅱ活性的检测。结果Rofecoxib能明显的抑制环氧化酶Ⅱ的活性。结论该方法可以用于环氧化酶Ⅱ抑制剂的筛选。
Abstract
OBJECTIVE To establish the assay model of cyclooxygenase-2 activity and provide the screening METHOD of NSAIDs.METHODS Observations were made that cyclooxygenase-2 was induced in the exist of FCS.The inhibitory potency of the compounds was evaluated by determining the concentration of PGE2.Rofecoxib employed as the reference drug was used to evaluate whether this method was available to determine the inhibitory potency of cyclooxygenase-2.RESULTS The results proved that rofecoxib can inhibit the activity of cyclooxygenase-2.CONCLUSION The in vitro method can be used to screen the cyclooxygenase-2 inhibitor.
关键词
环氧化酶 /
PGE2 /
NIH3T3 /
筛选模型
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Key words
cyclooxygenase 2 /
PGE2 /
NIH 3T3 /
screen model
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参考文献
［1］O'Bannion MK,Sadowshi HB,Winn V,et al.A serum and glucocorticoidregulated 4kilobase mRNA encodes a cycloocygenaserelated protein［J］.J Biol Chem,1991，266：23261.
［2］deWitt DL,Meade E,Smith WL.PGH sysnthase isoenzyme selectivity:the potential for safer nonsterodidal antiinflammatiory drugs［J］.Am J Med，1993，95：408.
［3］Shitashige M,Morita I,Murota S.Different substrate utilization between prostaglandin endoperoxide H synthase1 and 2 in NIH 3T3 fibroblasts［J］.Biochimica Biophysica Acta，1998,1389(1)：57.
［4］胡玉芳，程桂芳.环氧化酶选择性抑制剂筛选模型的建立［J］.药学学报，2000，35(5)：343.
［5］Leval XD,Delarge J,Devel P,et al.Evaluation of classical NSAIDs and COX2 selective inhibitors on purified ovine enzymes and human whole blood［J］.Prostaglandins Leukotrienes Essent Fatty Acids,2001，64：211.
［6］Brock T,McNish R,PetersGolden M.Arachidonic acid is preferentially metabolized by cyclooxygenase2 to prostacyclin and prostaglandin E2［J］.J Biol Chem,1999,274(17):11660.
［7］Penglis P,Cleland L,Demasi M,et al.Differential regulation of prostaglandin E2 and thromboxane A2 production in human monocytes:implications for the use of cyclooxygenase inhibitors［J］.J Immunol,2000,165：1605.
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脚注
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基金
南京医科大学应用毒理学重点实验室开放基金（K2034）和江苏省自然科学基金（BK99099）
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