灌胃“益气养阴方”大鼠的血清诱导白血病K562细胞凋亡的实验研究

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1、山东中医药大学硕士学位论文灌胃“益气养阴方”大鼠的血清诱导白血病K562细胞凋亡的实验研究姓名：臧运华申请学位级别：硕士专业：中西医结合基础指导教师：李震2003.4.20提要'占01163目的：探讨“益气养阴方”治疗白血病的作用机理。方法：运用血清药理学方。●__----_-。一法孵育人白血病细胞系K562细胞，通过荧光光度测定法、流式细胞术、免疫组化法分别检测灌胃大、中、小剂量的“益气养阴方”大鼠的血清对膜流动性、细胞周期和细胞凋亡及bcl一2、突变型p53基因表达的影响。／结果：小剂量组对细胞膜流动L性无明显影响。而大剂量组使K562细胞膜流动性增高，表

2、现出与注射砒霜大鼠的血清和灌胃羟媾脲大鼠的血清相同的趋势：灌胃三利哜4量“益气养阴方”大鼠的血清均能使K562细胞s期细胞百分数增加，Go—GI期细胞百分数减少，与注射砒霜大鼠的血清影响结果相似。三种剂量的血清均能使K562细胞bcl-2和突变型p53基因表达减少，它们都能诱导K562细胞调亡。厂结论：灌胃大、-I-、小剂量“益气养阴方”大鼠的血清均能使K562细胞阻滞予s期．并且诱导K562细胞调亡。这种作用可能与其使K562细胞膜流动性增高及bcl_2和突变型p53基冈表达减少有关。关键词益气养阴方Experimentstudyonserumofthera

3、tsthatAreGivenViQiYangyinformulainducestheApoptosisofHumanLeukemiacelllinesK562Speciality：BasicresearchofintegratedTCMandwesternmedicineAuthor：ZangYunhuaTutor：Prof．1iZhenAbstractObjectiveToexplorethemechamismoftreatingleukemiawithYiqiYangyinformula．MethodsWehatchedcelllinesK562withh

4、umanleukemiabymeansofserologicalpharmacology．Atthesametime，YiqiYangyinformulaisgiventotherats．Andthen，weobserverats’serum’seffectsonmembranefluidity，cellcycleandbcl-2geneandmutantp53geneexpressionofcelllinesK562byfluorophotometry，flowcytomeuyandimmunohistochemic．ResultsSerumoftherat

5、sgivenYiqiYangyinformulaatlowerdrugconcentrationshaslittleeffectonthemembranefluidity．ButathigherdrugconcentrationsitcanincreasemembranefluidityofcelllinesK562，thiseffecthasthesametendencytoserumoftheratsgivenarsenicalagentsandthatgivenHU．SerumoftheratsgivenYiqiYangyinformulacanincr

6、easemembranefluidityandSphasecells’percentageofcelllinesK562anddecreaseGO-Glphasecells’percentage，whichissimilartotheeffectoftherats’serumthataregivenarsenicalagents．SerumoftheratsgivenYiqiYangyinformuladown·regulatestheexpressionofbcl-2geneandmutantp53gene，andinadditionitinducesthe

7、apoptosisofthecelllinesK562．ConclusionWithserumoftheratsgivenYiqiYangyinformula，SphasecellsofcelllinesK562increased，andthenapoptoticcellsincreasedwithadiminutionofSphasecells．ThismechanismmayrelatetothefunctionthatincreasemembranefluidityofcelllinesK562anddown·regulatetheexpressiono

8、fbcI·2andmutantp53g