线粒体途径在大黄素促人混合培养淋巴细胞凋亡中的作用.pdf

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1、现代肿瘤医学2015年06月第23卷第11期MODERNONc0L0GY，Jun．2015，VOL23，NO．11·1495·线粒体途径在大黄素促人混合培养淋巴细胞凋亡中的作用沈乃营，刘昌，何盟国，曲凯，张天政EfectofthemitochondrialsignalingpathwayonEmodin——inducedapoptosisinhumanmixedlymphocytecultureShenNaiying，LiuChang，HeMengguo，QuKai，ZhangTianzhengDepartmentofHepatobiliarySurge

2、ry，ShaanxiXianyang215Hospital，ShaanxiXi(myang712000，China；DepartmentofHepatobiliarySurgery，theFirstAffiliatedHospitalMedicalSchoolofnJiaotongUniversity，ShaanxiXihn710061，China．【Abstract】0bjective：ToinvestigatetheeffectofmitoehondrialsignalingpathwayonEmodin—inducedapoptosisinhuma

3、nmixedlymphocyteculturemode1．Methods：Themodelofhumanmixedlymphocyteculturewasestablishedandlymphocyteswereisolatedfromtheperipheralbloodsamplesofdifferenthealthyadultvolunteers．ToinvestigatetheimmunosuppressiveeffectofEmodin．Emodinwasadministeredwithdifferentconcentrationsat1i．~m

4、ol／L，101~mol／L，1001~mol／Lfor24，48，72h．Attheendofculture，theapoptoticrateoflymphoeytesandthelevelofreactiveoxygenspecies(ROS)andmitoehondrialmembranepotential(△m)wereevaluatedwithfloweytometry．Proteinexpres-sionsofeytosoliccytochromeC，caspase一3andcaspase一9wereanalyzedbyWesternblot

5、．InthehighconcentrationEmodingroup，lymphocyteswereculturedwithreducedglutathione(GSH)andEmodinfor48h，GSHcanremovein-tracellularROS．TheapoptoticrateoflymphocytesandthelevelsofROSand△蛳werealsoevaluatedbyfloweytom-etry．ProteinexpressionsofBcl一2andBaxwereanalyzedbyWesternblot．Results

6、：WiththeenhancedconcentrationofEmodin，theinhibitionofproliferationandincreaseofapoptoticratewereobserved．TheresultsfurthershowedgenerationofROS，disruptionof△m，decreaseoftheBcl一2／BaxratioandcytosoliccytochromeC．Caspase一3andcaspase一9levelsalsoincreasedaftertheEmodintreatment．Thedec

7、lineintheBcl一2／BaxratioandtheincreaseofROS，cytosoliccytochromeCandcaspase一3andcaspase一9levelswereconsistentwiththeincreaseofthelympho—cyteapoptoticratio．IntheGSHgroup，thelevelofROSdeclined，andtheapoptoticratealsoreducedcomparedwiththehighconcentrationEmodingroup(P<0．05)，however，t

8、heAgtmenhancedcomparedwithcontrolgroup(P