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1、益气活血方药对老年大鼠肝脏衰老相关基因表达的影响论文黎志萍，赵伟康，徐品初，陈方敏，金国琴，姚明忠【关键词】肝细胞［摘要］目的：观察益气、活血方药对老年大鼠肝脏衰老相关基因表达的影响。方法：以自然衰老大鼠为衰老模型，采用RTPCR、ethods:Arecipeforreplenishingqiandarecipeforactivatingbloodinisteredtoagingratsrespectively,andtheeffectsoftheaboverecipesontheexpressionsofsenescencerelatedgenes(p16,p21,P
2、A,cyclinD1andcyclinE)inedbyRTPCRandSF)和40μl混合蛋白酶抑制plete液(pleteEDTAfreeproteaseinhibitorcocktailtablet,1片加1ml水），4℃10000r/min离心10min，取上清液于－70℃冷冻保存。24h后10000r/min离心10min，取上清液分装，－70℃冷冻保存。Lododecylsulfatepolyacrylamidegelelectrophoresis,SDSPAGE)和免疫印迹反应，用SupersignalECL发光剂荧光显色X光胶片曝光，观察蛋白条带。1.5图
3、像处理及统计分析RTPCR、:Marker;1:Youngcontrolgroup;2:Untreatedgroup;3:Qibenefitingrecipetreatedgroup;4:Bloodactivatingrecipetreatedgroup图2cyclinD1、PA和cyclinE琼脂糖凝胶电泳（略）Fig2AgarosegelelectrophoresisofcyclinD1,PAandcyclinEM:Marker;1:Youngcontrolgroup;2:Untreatedgroup;3:Qibenefitingrecipetreatedgroup
4、;4:Bloodactivatingrecipetreatedgroup表1益气、活血方药对大鼠肝脏衰老相关基因mRNA表达的影响（目的基因/βactin）（略）Tab1EffectsofqibenefitingandbloodactivatingrecipesonmRNAexpressionsofp16,p21,PA,cyclinD1andcyclinEgenesintheliverofrats(targetgene/βactin)*P0.05,**P0.01,vsyoungcontrolgroup;△△P0.01,vsuntreatedgroup表2益气、活血方药对
5、大鼠肝脏衰老相关基因蛋白表达的影响（目的基因/βactin）（略）Tab2Effectsofqibenefitingandbloodactivatingrecipesonproteinexpressionsofp16，p21，PA，cyclinD1andcyclinEgenesintheliverofrats(targetgene/βactin)**P0.01,vsyoungcontrolgroup;△△P0.01,vsuntreatedgroup图3p16、p21、PA、cyclinD1和cyclinE蛋白印迹杂交结果（略）Fig3K.Agedependentmodi
6、ficationsofgeneexpressioninhumanfibroblasts［J］.CritRevEukaryotGeneExpr,1998,8(1):4380.2McConnellBB,StarborgM,BrookesS,etal.Inhibitorsofcyclindependentkinasesinducefeaturesofreplicativesenescenceinearlypassagehumandiploidfibroblasts［J］.CurrBiol,1998,8(6):351354.3SmithJR,PereiraSmithOM.Rep
7、licativesenescence:implicationsforinvivoagingandtumorsuppression［J］.Science,1996,273(5271):6367.4杨美玲,周吉燕,徐维蓉.益气、补肾、活血、健脾四组方药对老年大鼠肝脏形态学变化影响的比较研究［J］.中国中医药科技,1996,3(6):2728,34.5姚明忠,顾文聪.固真方对老年大鼠肝脏原癌基因和抗癌基因表达的影响［J］.中国中西医结合杂志,1997,17(基础理论特辑):67.6NakajimalijimaS,HamadaH,ReddyP,e
