GnRHa和孕三烯酮对异位子宫内膜细胞生长及凋亡的影响

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1、GnRHa和孕三烯酮对异位子宫内膜细胞生长及凋亡的影响作者：王春凤,姜卫国单位：山东大学医学院山东省千佛山医院妇产科，济南【摘要】目的探讨GnRHa及孕三烯酮对体外培养的异位子宫内膜间质(ES)细胞生长和凋亡的影响，及两种药物作用后基质金属蛋白酶3(MMP3)在ES细胞表达的变化。方法以不同浓度的GnRHa和孕三烯酮对体外培养的ES细胞分别进行处理，倒置相差显微镜下观察细胞的生长状态，免疫组化法鉴定间质细胞。采用四甲基偶氮唑蓝(MTT)比色法检测药物作用0～60?h的细胞抑制情况，并绘制细胞生长曲线；采用流式细胞仪检测GnRHa

2、和孕三烯酮作用48h后异位内膜间质细胞周期和凋亡率的变化。采用免疫组化法检测不同浓度GnRHa和孕三烯酮作用48h后MMP3在异位内膜表达的变化。结果GnRHa和孕三烯酮可抑制细胞的生长增殖，并呈时间和剂量依赖方式。药物作用48?h后，各组细胞均发生细胞周期阻滞，G1期细胞增加，S期细胞减少，各组凋亡率明显升高(P<0.05)；免疫组织化学表明，两种药物均可降低异位子宫内膜MMP3的表达；且GnRHa诱导异位子宫内膜细胞的凋亡、降低其MMP3表达的作用强于孕三烯酮(P<0.05)。结论GnRHa和孕三烯酮均可明显抑

3、制体外培养的ES细胞的生长增殖，诱导其凋亡，同时降低MMP3的表达，且GnRHa上述作用的效果更加明显。【关键词】子宫内膜异位症；GnRHa；孕三烯酮；细胞周期；细胞凋亡inectopicendometrialstromacellsWANGChunfeng,JIANGWeiguo(DepartmentofGynecologyandObstetrics,ShandongQianfoshanHospital,SchoolofClinicalMedicine,ShandongUniversity,Jinan250014，China)

4、Abstract:ObjectiveToinvestigatetheeffectsofGnRHaandGestrinoneongrowth,proliferationandapoptosisinisolatedectopicendometrialstromacellsinvitro,aswellasexpressionofMMP3inducedbyGnRHaandGestrinone.MethodsAxenicectopicendometriotictissueswereisolatedandculturedinvitro.The

5、ntheculturedendometrialstromacellswereobservedbyaninvertedmicroscopeandidentifiedbyvimentinimmunohistochemicalstaining.TheywereculturedandexposedtoGnRHaandGestrinoneatdifferentdoses.Theinhibitionofthecellsin60hourswasdeterminedbymethylthiazolyltetrazolium(MTT)assay,and

6、acellgrowthcurvewasmade.At48hoursthecellcycleandapoptosisweremonitoredbyflowcytometry(FCM),andexpressionofMMP3inisolatedectopicendometrialstromacellswasanalyzedbyimmunohistochemistry.ResultsGnRHaandGestrinoneobviouslyblockedthecellcycleofectopicendometrialstromacellsw

7、iththenumberofcellsincreasedattheG1stageanddecreasedattheSstage.Theapoptoticrateofendometrialstromacellswassignificantlyincreased(P<0.05),whileexpressionofMMP3inducedbyGnRHaandGestrinonewasobviouslydecreased(P<0.05),buttheapoptosiswasmoresignificantlyenhancedand

8、expressionofMMP3wasmoreobviouslydecreasedbyGnRHathanbyGestrinone.ConclusionsGnRHaandGestrinonecansignificantlyinhibi