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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,andtheeffectsoftheaboverecipesontheexpressionso
2、fsenescencerelatedgenes(p16,p21,PA,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。Lododecylsulfatepolyacrylamidegelelec
3、trophoresis,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) Fig2Agarosegelelectro
4、phoresisofcyclinD1,PAandcyclinE M:Marker;1:Youngcontrolgroup;2:Untreatedgroup;3:Qibenefitingrecipetreatedgroup;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) Tab1EffectsofqibenefitingandbloodactivatingrecipesonmRNAe
5、xpressionsofp16,p21,PA,cyclinD1andcyclinEgenesintheliverofrats(targetgene/βactin) *P0.05,**P0.01,vsyoungcontrolgroup;△△P0.01,vsuntreatedgroup 表2益气、活血方药对大鼠肝脏衰老相关基因蛋白表达的影响(目的基因/βactin)(略) Tab2Effectsofqibenefitingandbloodactivatingrecipesonproteinexpre
6、ssionsofp16,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.Agedependentmodificationsofgeneexpressioninhumanfibroblasts〔J〕.CritRevEu
7、karyotGeneExpr,1998,8(1):4380. 2McConnellBB,StarborgM,BrookesS,etal.Inhibitorsofcyclindependentkinasesinducefeaturesofreplicativesenescenceinearlypassagehumandiploidfibroblasts〔J〕.CurrBiol,1998,8(6):351354. 3SmithJR,PereiraSmithOM.Replicativesenescenc
8、e: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. 6