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1、表觀遺傳修飾在大鼠BMSCs向BSMCs分化中調(diào)控機(jī)制的研究徐夢(mèng)遙，王延洲，徐惠成(400038重慶，第三軍醫(yī)大學(xué)西南醫(yī)院婦產(chǎn)科)、(國(guó)家自然科學(xué)基金青年科學(xué)基金，項(xiàng)目編號(hào)30801234)[摘要]目的通過(guò)骨髓間充質(zhì)干細(xì)胞(bonemarrowmesenchymalstemcells,BMSCs)與膀胱平滑肌細(xì)胞(bladdersmoothmusclecells,BSMCs)接觸共培養(yǎng)探討組蛋白乙?；瘜?duì)BMSCs分化能力的影響。方法 分別采用密度梯度離心法和膠原酶消化法培養(yǎng)大鼠BMSCs及BSMCs至第3代，并對(duì)BMSCs進(jìn)行了鑒定；以單獨(dú)培養(yǎng)的B
2、MSCs作為對(duì)照組，接觸共培養(yǎng)的BMSCs為實(shí)驗(yàn)組；運(yùn)用免疫熒光化學(xué)染色法分別對(duì)2組BMSCs中的平滑肌標(biāo)志性蛋白平滑肌α肌動(dòng)蛋白(α-smoothmuscleactin,α-SMA)、平滑肌肌球蛋白重鏈(smoothmuscle-myosinheavychain,SM-MHC)、calponin進(jìn)行檢測(cè)，并用微球菌核酸酶(MicrococcalNuclease)對(duì)平滑肌三個(gè)標(biāo)志性基因的啟動(dòng)子區(qū)進(jìn)行了染色質(zhì)開(kāi)放性的檢測(cè)，同時(shí)運(yùn)用了ChIP技術(shù)分析了α-SMA、SM-MHC啟動(dòng)子區(qū)組蛋白乙?；綄?duì)BMSCs向平滑肌分化的影響。結(jié)果 大鼠原代BMSC
3、s流式檢測(cè)顯示CD29表達(dá)陽(yáng)性,陽(yáng)性率分別為99.8%；CD31、CD45表達(dá)陰性，陽(yáng)性率為6.30%、1.04%；免疫熒光顯示平滑肌3種標(biāo)志性蛋白的表達(dá)均隨時(shí)間的延長(zhǎng)而增加；MNase顯示實(shí)驗(yàn)組的BMSCs啟動(dòng)子區(qū)染色質(zhì)對(duì)核酸酶的敏感性明顯高于對(duì)照組，ChIP顯示實(shí)驗(yàn)組中BMSCs平滑肌標(biāo)志性基因α-SMA、SM-MHC、Calponin啟動(dòng)子區(qū)H4乙?；捷^未分化前明顯增加(P<0.05)。結(jié)論組蛋白乙?；潭鹊脑黾哟龠M(jìn)了BMSCs向BSMCs分化。[關(guān)鍵詞] 骨髓間充質(zhì)干細(xì)胞;膀胱平滑肌細(xì)胞;組蛋白乙?；?啟動(dòng)子Theregulatorym
4、echanismofepigeneticmodificationondifferentiationofratbonemarrowmesenchymalstemcellstosmoothmusclecellsXuMengyao,WangYanzhou,XuHuicheng(DepartmentofGynecologyandObstetricsSouthwestHospitalThridMilitaryMedicalUniversity,Chongqing400038China)[Abstract]ObjectiveMesenchymalstemcel
5、lsinthebonemarrow(bonemarrowmesenchymalstemcells,BMSCs)andbladdersmoothmusclecells(bladdersmoothmusclecells,BSMCs)coculturehistoneacetylationBMSCsdifferentiationcapacity.Method:Densitygradientcentrifugtionandcollagenasedigestionmethodwereusedtotraintherats'BMSCsandBSMCstothe3r
6、dgeneration,andidentifytheBMSCs.BMSCsculturedaloneasthecontrolgroup,whichcontactstheco-culturedBMSCsfortheexperimentalgroup.UsingimmunefluorescencestainingonthesmoothmuscleofthetwogroupswhichisBMSCslandmarkproteinonthesmoothmuscleαactin(α-smoothmuscleactin,α-SMA)andsmoothmuscl
7、emyosinheavychain(smoothmuscle-myosinheavychain,SM-MHC)whichwillbedetected,andthepromoterregionsofthethreemarkergenesinsmoothmusclewithmicrococcalnuclease(MicrococcalNuclease)chromatinopenadetection,whiletheuseofChIPtechnicalanalysisofα-SMA,SM-MHCthepromoterregionsofgroupprote
8、inacetylationlevelsofBMSCstosmoothmuscledifferentiation.Resul
