【摘要】 目的:探讨Notch1/Hes1信号通路能否通过调控CCAAT/增强子结合蛋白α（C/EBPα）的表达从而影响肺泡Ⅱ型上皮细胞（AECⅡ）的增殖与分化功能。方法:体外培养人AECⅡ,将细胞随机分为对照组、激活剂组（加入Notch通路激活剂Jagged1蛋白500μg/L）和抑制剂组（加入Notch通路抑制剂DAPT 10μmol/L）,于干预后24 h收获各组细胞。采用RT-qPCR和Western blot法分别检测Notch1、Hes1及C/EBPα的mRNA与蛋白表达水平;CCK-8法检测细胞活力;细胞计数检测细胞增殖;流式细胞术检测细胞周期及分化。结果:与对照组相比,激活剂组Notch1、Hes1和C/EBPα的mRNA和蛋白表达显著增加（P<0.05）,促进AECⅡ从S期进入G₂/M期,增殖增加而分化减少（P<0.05）;抑制剂组Notch1、Hes1和C/EBPαmRNA和蛋白表达水平明显降低（P<0.05）,AECⅡ被阻滞于G₀/G₁期,增殖减少而分化增加（P<0.05）。结论:Notch1/Hes1信号可调控C/EBPα表达并能影响AECⅡ增殖与分化。更多还原

【Abstract】 AIM: To investigate whether Notch1/Hes1 signaling pathway regulates the expression of CCAAT/enhancer binding protein alpha（C/EBPα）, thus affecting the proliferation and differentiation of type Ⅱ alveolar epithelial cells（AECⅡ）. METHODS: Human AECⅡ were cultured in vitro, and randomly divided into control group, activator group（adding Notch pathway activator Jagged1 protein, 500 μg/L） and inhibitor group（adding Notch inhibitor DAPT, 10 μmol/L）. AECⅡ in each group were collected after 24 h of intervention. The expression of Notch1, Hes1 and C/EBPα at mRNA and protein levels was determined by RT-qPCR and Western blot. The cell proliferation ability of the AECⅡ was measured by living cell counting and CCK-8 assay. The cell cycle distribution and differentiation of the AECⅡ were analyzed by flow cytometry. RESULTS: Compared with control group, the mRNA and protein expression levels of Notch1, Hes1 and C/EBPα were significantly increased in activator group（P<0.05）, AECⅡ entered G₂/M phase from S phase, the proliferation of AECⅡ was increased, and the differentiation of AECⅡ was reduced（P<0.05）. The mRNA and protein expression levels of Notch1, Hes1 and C/EBPα were significantly reduced in inhibitor group（P<0.05）, the cell cycle of AECⅡ cells was arrested in G₀/G₁ phase, the proliferation of AECⅡ cells was reduced, and the differentiation of AECⅡ cells was increased（P<0.05）. CONCLUSION: Notch1/Hes1 signaling pathway regulates the expression of C/EBPα and affects the proliferation and differentiation of AECⅡ.更多还原