Deltex-1对骨髓间充质干细胞增殖及其向平滑肌细胞分化的影响  

Deltex-1 overexpression suppresses proliferation and promotes differentiation of bone marrow mesenchymal stem cells into smooth muscle cells in vitro

作　　者：王荥[1] 王延洲 刘禄斌[1] 徐惠成 

机构地区：[1]重庆市妇幼保健院,重庆400021 [2]陆军军医大学(第三军医大学)第一附属医院妇产科,重庆400038

出　　处：《第三军医大学学报》2018年第2期109-115,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(30772309)~~

摘　　要：目的探讨Deltex-1对骨髓间充质干细胞(bone marrow mesenchymal stem cells,bMSCs)增殖及其向平滑肌细胞(smooth muscle cells,SMCs)分化的影响。方法分离培养大鼠骨髓bMSCs,采用流式细胞仪检测鉴定。通过Deltex-1过表达腺病毒载体p Ad/Deltex-1感染bMSCs后,采用CCK-8(cell count kit-8)法检测Deltex-1过表达对bMSCs增殖的影响;将未感染、经Deltex-1病毒感染及经空病毒感染的bMSCs与SMCs共培养,采用免疫荧光细胞化学、RT-PCR及Western blot分别检测其bMSCs中SMCs标志物平滑肌肌球蛋白重链(smooth muscle myosin heavy chain,SM-MHC)及Notch信号通路相关因子Notch-1的表达,单纯培养的bMSCs、SMCs的相同检测作正常对照。结果成功分离、培养大鼠bMSCs。CCK-8检测证实:与对照比较,经Deltex-1病毒感染的bMSCs生长减慢,48 h开始显得更为明显(P<0.05,P<0.01);免疫荧光细胞化学、RT-PCR及Western blot检测显示:与对照比较,经Deltex-1病毒感染的bMSCs与SMCs共培养后显著表达SMCs的标志物SM-MHC,较弱表达Notch信号通路相关因子Notch-1(P<0.01)。结论 Deltex-1过表达可抑制bMSCs的增殖,并促进其向SMCs分化。ObjectiveTo investigate the effect of Deltex1 overexpression on the proliferation of bone marrow mesenchymal stem cells （bMSCs） and their differentiation into smooth muscle cells （SMCs） in vitro. MethodsRat bMSCs were isolated, cultured and identified. The effect of Deltex1 overexpression on the proliferation of bMSCs infected with the adenovirus vector pAd/Deltex1 was observed using cell count kit8 （CCK8）. In a coculture system with SMCs, bMSCs infected with pAd/Deltex1 or an empty viral vector or with no treatment were examined for the expression of smooth muscle myosin heavy chain （SMMHC, a marker of SMCs） using immunofluorescence cytochemistry staining, RTPCR and Western blotting, with bMSCs and SMCs without treatment in separate cultures as normal controls. ResultsCompared with the control cells, bMSCs infected by the adenovirus vector pAd/Deltex1 exhibited significantly lowered growth rate, which was especially obvious at 48 h after the infection （P〈005, 0.01）. The results of immunofluorescence cytochemistry, realtime PCR and Western blotting all showed that bMSCs infected with the Deltex1 vector and cocultured with SMCs expressed a significantly increased level of SMMHC with a lowed expression of Notch1 （P〈001）. ConclusionDeltex1 overexpression causes suppression of bMSC proliferation and promotes the differentiation of bMSCs into smooth muscle cells in vitro.

关 键 词：Deltex-1 骨髓间充质干细胞 平滑肌细胞 细胞分化 

分 类 号：R322.74[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]