中文摘要：

目的：探讨垂体腺苷酸环化酶激活肽（PACAP）的PAC1受体特异激动剂maxadilan对人诱导多能干细胞（IPSCs）增殖的作用。方法：培养人IPSCs,利用RT-PCR和Western blotting检测IPSCs的PAC1受体;IP-SCs培养基中加入不同浓度的maxadilan作为实验组,未加入maxadilan作为对照组。CCK-8法检测maxadilan对IPSCs增殖的影响;流式细胞仪分析maxadilan对细胞周期的影响;染色体核型分析检测maxadilan对IPSCs核型的影响;real-time-qPCR和免疫荧光法从基因及蛋白水平检测maxadilan对IPSCs的多能性基因Nanog和OCT4的影响;real-time qPCR检测maxadilan对IPSCs及其形成胚体细胞的Nestin和PAX6基因的影响;RT-PCR检测maxadilan对IPSCs分化为三胚层能力的影响。结果：RT-PCR和Western blotting均显示IPSCs含有PAC1受体;CCK-8法显示100 nmol/L maxadilan组比对照组细胞增多16%（P〈0.05）;流式细胞仪分析显示孵育100 nmol/Lmaxadilan 3 h、6 h和9 h后IPSCs增殖指数为47.23%、59.70%、55.67%,与对照组37.00%相比,差异显著（P〈0.05）;染色体核型分析显示maxadilan处理过的IPSCs核型正常;RT-PCR、real-time qPCR和免疫荧光法显示maxadilan未影响IPSCs多能性。结论：人IPSCs存在PAC1受体,maxadilan对人IPSCs具有促进细胞增殖作用且没有影响IPSCs的多能性和染色体核型。

英文摘要：

AIM：To investigate the promoting effect of maxadilan,which specifically activate the type I receptor for pituitary adenylate cyclase-activating polypeptide（PACAP）,on the proliferation of human induced pluripotent stem cells（IPSCs）.METHODS： PACAP type I（PAC1） receptor in IPSCs was detected by RT-PCR and Western blotting.maxadilan at various concentrations was added to the medium of IPSCs as experimental groups.The medium in control group was without maxadilan treatment.The effect of maxadilan on the proliferation of IPSCs was measured with Cell Counting Kit-8（CCK-8）.The changes of cell cycle caused by maxadilan in IPSCs were analyzed by flow cytometry.The analysis of karyotype was carried out in IPSCs treated with maxadilan.Proteins and gene expression levels of both Nanog and OCT4 in IPSCs treated with maxadilan were detected by real-time quantitative polymerase chain reaction（real-time-qPCR） and immunofluorescence.The gene expression levels of Nestin and PAX6 in both IPSCs treated with maxadilan and cells of embryonic body,which was birthed from IPSCs with maxadilan treatment,were detected by real-time qPCR.The ability of IPSCs treated with maxadilan differentiating into 3 embryonic layers was evaluated by analyzing the component of embryo using RT-PCR.RESULTS： The PAC1 receptor in IPSCs was identified by RT-PCR and Western blotting.Viability of the IPSCs with 100 nmol/L maxadilan treatment was increased by 16% compared with control group.The differences with statistical significance were found in the cell viability between 100 nmol/L maxadilan treatment group and control group（P〈0.05）.The average values of proliferation index（PI） in IPSCs with 100 nmol/L maxadilan treatment for 3 h,6 h and 9 h were 47.23%,59.70% and 55.67%,respectively,while that in control group was 37.00%.The differences with statistical significance were found in PI between 100 nmol/L maxadilan treatment for 3 h group,6 h group,9 h group and control group（P〈0.05）.Normal karyotype and unchanged plurip