IGF-1抑制妊娠合并糖尿病诱导的鼠胚胎细胞凋亡

IGF-1inhibitstheapoptosisofmouseembryoinducedbydiabetes投稿时间：2008-10-18最后修改时间：2009-2-13稿件编号：20080711中文关键词：妊娠合并糖尿病鼠胚胎体外培养IGF-1凋亡英文关键词：pregnancyassociatedwithdiabetesmouseembryosinvitroculture（IVC）IGF-1apoptosis基金项目：国家自然科学基金资助项目（30600224,30700438,30771135）,中国博士后科学基金（20060390886）,湖南省自然科学基金（06JJ30014）,湖南省科技计划（2008FJ3138）和湖南省教育厅科研资助项目（07B008）作者单位罗晓敏长沙医学院基础医学系，长沙410219；中南大学湘雅医学院组织学与胚胎学系，长沙410013邹海燕长沙医学院基础医学系，长沙410219；中南大学湘雅医学院组织学与胚胎学系，长沙410013张粗心洗笱嫜乓窖г鹤橹в肱咛パ担ど410013潘爱华中南大学湘雅医学院组织学与胚胎学系，长沙410013尹芳长沙医学院基础医学系，长沙410219魏苗中南大学湘雅医学院组织学与胚胎学系，长沙410013杨静中南大学湘雅医学院组织学与胚胎学系，长沙410013黄河中南大学湘雅医学院组织学与胚胎学系，长沙410013摘要点击次数：283全文下载次数：145中文摘要：为研究妊娠合并糖尿病对孕妇及胎儿产生危害的机制，构建妊娠合并糖尿病的昆明小鼠动物模型，检测不同浓度葡萄糖对体外培养胚泡细胞生长的影响．观察不同浓度胰岛素样生长因子-1（insulin-likegrowthfactor-1,IGF-1）对体外高糖环境胚泡细胞发育的影响，利用细胞核DNA双染实验观察不同浓度IGF-1作用下胚泡细胞的凋亡．采用实时定量PCR（RT-PCR）分析不同凋亡相关基因在体外培养胚泡中的表达情况．结果显示，随着葡萄糖浓度的增加，胚泡细胞总数减少，高浓度葡萄糖（≥30mmol/L）则能显著性抑制胚泡细胞的生长（P<0.01）．RT-PCR检测发现妊娠合并糖尿病小鼠的胚泡igf-1表达下调，且与葡萄糖的浓度成正相关．凋亡相关基因bcl-2和bcl-xl的表达随着体外IGF-1培养浓度的增加而表达上调，而p53基因和凋亡相关基因Bax的表达则下调．细胞凋亡实验显示，随着IGF-1浓度的增加，体外培养胚泡细胞的凋亡逐渐降低，当IGF-1浓度达到100μg/L时，几乎未发现细胞凋亡．因此，高血糖能抑制胚泡细胞的生长发育，导致igf-1的表达下调，而IGF-1能抑制胚泡细胞的凋亡，有利于胚泡细胞的生长发育．英文摘要：Inordertoinvestigatethemechanismsofpregnancyassociatedwithdiabetesharmtogravidaandfetal,TheKunmingmicemodelsforpregnancyassociatedwithdiabetesweresimulated,Theeffectsofglucoseindifferentconcentrationswasdetectedonthegrowthofembryocellsinvitroculture.TheeffectsofIGF-1indifferentconcentrationswasdeterminedonthedevelopmentofblastocystinhyperglycemicconditions（30.0mol/Lglucose）invitro,aswellastheapoptosisofembyrocellsbyusingnuclearDNAdouble-dyedassays.Moreover,theexpressionofigf-1andothergenesassociatedwithapoptosisinvitroembryowasdetctedbyReal-timequantitativePCR.Theresultsshowedthatthetotalcellnumberofblastocystfelloffalongwiththeincreasingofglucoseconcentration.Highconcentrationsofglucose（≥30mmol/L）couldsignificantlyinhibitthegrowthofembryocells（P<0.01）.Theresultsofreal-timequantitativePCRshowedthattheexpressionofigf-1wasdown-regulatedinmouseblastocystwithpregnancyassociatedwithdiabetes,andpositivelycorrelatedwiththeconcentrationofglucose.Theexpressionsofapoptosis-relatedgenesbcl-2andbcl-xlwereup-regulatedalongwiththeincreasingofIGF-1concentration,whilethep53geneandtheapoptosis-relatedgeneBaxweredown-regulated.TheembryocellsapoptosishadbeengraduallyreducedwiththeincreasingofIGF-1concentrationinvitro,andintheIGF-1concentrationof100μg/L,therewasalmostnoapoptosisinembryocells.Theseexperimentsdemonstratedthathyperglycemicconditionsinvitrocaninhibitthegrowthanddevelopmentoftheembryocellsresultinginthedown-regulatedexpressionofigf-1,andIGF-1couldinhibittheapoptosisofblastocystandbebenefitforthegrowthofblastocyst.