Maxadilan 对兔角膜上皮细胞的促增殖作用  

Maxadilan on proliferation of corneal epithelial cells in rabbits

作　　者：招志毅[1,2] 陈建苏[1,2] 余榕捷[3] 谭美华[1,2] 李红阳 刘晓飞[3] 王静静[3] 

机构地区：[1]暨南大学附属第一医院眼科 [2]暨南大学医学院眼科研究所再生医学教育部重点实验室,广东省广州市510632 [3]暨南大学生物医学工程研究所,广东省广州市510632

出　　处：《眼科新进展》2011年第10期901-903,911,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30973244) ;暨南大学 211项目、中央高校基本科研业务费专项资金资助(编号:21609407)~~

摘　　要：目的探讨垂体腺苷酸环化酶激活肽(pituitary adenylate cyclase activating polypeptide,PACAP)的PAC1受体特异激动剂Maxadilan对角膜上皮细胞的促增殖作用。方法分离新西兰大白兔角膜上皮细胞,并在含体积分数10%胎牛血清的DMEM培养基中培养。利用逆转录聚合酶链反应(RT-PCR)检测兔角膜上皮细胞PACAP的PAC1受体;细胞增殖实验采用CCK-8法检测,角膜上皮细胞培养基中分别加入0.01μmol.L-1、0.03μmol.L-1、0.05μmol.L-1、0.10μmol.L-1、0.15μmol.L-1Maxadilan作为实验组,未加入Maxadilan作为对照组,检测不同浓度的Maxadilan对角膜上皮细胞增殖的影响;流式细胞仪分析0.10μmol.L-1Maxadilan对细胞周期的影响。结果 RT-PCR检测显示兔角膜上皮细胞含有PACAP的PAC1受体;CCK-8法检测显示0.05μmol.L-1、0.10μmol.L-1、0.15μmol.L-1Maxadilan实验组分别比对照组细胞增多29.1%、41.5%、32.5%,与对照组相比差异有统计学意义(P分别为0.001、0.000、0.000);流式细胞仪分析显示0.10欁μmol.L-1Maxadilan组细胞增殖指数平均值为35.98%,与对照组细胞细胞增殖指数平均值29.48%相比,差异有统计学意义(P=0.014)。结论角膜上皮细胞存在PACAP的PAC1受体,Maxadilan对角膜上皮细胞有促增殖作用。Objective To investigate the effects of Maxadilan,which can specifically activate the type I receptor of pituitary adenylate cyclase activating polypeptide（PACAP）,on promoting the proliferation of corneal epithelial cells（CEC）.Methods CEC was isolated from New Zealand white rabbits and cultured in Dulbecco＇s modified Eagle medium containing volume fraction of 10% fetal calf serum.PACAP type I（PAC1） receptor in CEC was detected with reverse transcriptase polymerase chain reaction（RT-PCR） assay.The effect of proliferation of CEC caused by Maxadilan was measured with cell counting Kit-8（CCK-8） assay.Maxadilan at different concentrations of 0.01 μmol·L-1,0.03 μmol·L-1,0.05 μmol·L-1,0.10 μmol·L-1,0.15 μmol·L-1 were added to media of CEC as experimental groups,and CEC without the addition of Maxadilan was as control group.Different concentrations of Maxadilan on CEC were detected.Variance of cell cycle induced by 0.10 μmol·L-1 Maxadilan in CEC was analyzed with flow cytometer.Results The PAC1 receptor in CEC was identified after RT-PCR assay.Compared with control group,viability of cells in CEC with 0.05 μmol·L-1,0.10 μmol·L-1 and 0.15 μmol·L-1 Maxadilan experimental groups were more than 29.1%,41.5% and 32.5% respectively.Significant differences were found（P=0.001,P=0.000,P=0.000） respectively.The average value of proliferation index（PI） in CEC with 0.10 μmol·L-1 Maxadilan experimental group was 35.98% while that of control group was 29.48% using flow cytometer assay.Significant difference was found（P=0.014）.Conclusion PAC1 receptor of PACAP can be found in CEC,and Maxadilan can promote its proliferation.

关 键 词：MAXADILAN 角膜上皮细胞 细胞增殖 兔 

分 类 号：R772.2[医药卫生—眼科]