富血小板血浆对兔来源脂肪间充质干细胞wnt3基因和klotho基因表达的影响  

Effect of platelet-rich plasma on wnt3 gene and klotho gene of adipose-derived stem cells of rabbit

作　　者：曲辉[1] 胡刚[1] 康乐[1] 李翔[1] 秦宏智[1] 

机构地区：[1]大连医科大学附属第一医院整形科,116024

出　　处：《中华医学美学美容杂志》2015年第1期45-48,共4页Chinese Journal of Medical Aesthetics and Cosmetology

摘　　要：目的 探讨自体富血小板血浆（PRP）对兔来源脂肪间充质干细胞（ADSCs） wnt3基因和klotho基因启动表达的影响.方法 获取新西兰大白兔附睾脂肪组织中的脂肪干细胞,采用形态学及诱导分化进行细胞鉴定,将细胞培养至第4代,取新西兰大白兔腹主动脉血通过传统2次离心法制备PRP、贫血小板血浆（platelet-poor plasma,PPP）,将脂肪干细胞在含PRP（实验组）、含PPP（对照组）及空白全培（空白组）各5％的培养液中分别培养24、48、72h,将各组细胞裂解提取总RNA,做RT-PCR检测2种目的基因wnt3和klotho启动情况.结果 （1）提取的原代兔脂肪干细胞随着细胞增殖,呈梭形漩涡状生长.（2）分别用油红O染色和茜素红染色,均能成功着色.（3）逆转录-聚合酶链反应（RT-PCR）结果显示,实验组wnt3基因和klotho基因量明显增多（P＜0.05）.结论 通过分离、体外培养所得的细胞经形态学和多向诱导分化,确定为兔来源ADSCs.PRP可促进兔脂肪干细胞增殖并明显提高wnt3基因和klotho基因的表达.Objective To investigate the effect of platelet-rich plasma （PRP） on starting of wnt3 gene and klotho gene of adipose-derived stem cells （ADSCs） of rabbit.Methods Epididymal adipose tissue stem cells were obtained from New Zealand white rabbits,and the cells identified by morphology and inducing differentiation,and the cells were cultured to the fourth generation,PRP and PPP （platelet-poor plasma） were prepared by traditional centrifugal method from abdominal aortic of rabbit; ADSCs were cultured in culture medium containing PRP （experimental group）,PPP （control group） and all medium （blank group） for each 5％ for 24h,48h and 72h.Cells of each group were dissociated and total RNA extracted.Effects of the starting of wnt3 gene and klotho gene were detected by RT-PCR.Results Primary ADSCs of rabbit grew in the way of long spindle swirly.The results of oil red O and alizarin red staining of the ADSCs were positive.Expression of wnt3 gene and klotho gene in the experimental group significantly increased from the results of RT-PCR （P＜0.05）.Conclusions PRP can promote proliferation of the ADSCs of rabbit and increase the expression of wnt3 gene and klotho gene significantly.

关 键 词：富血小板血浆 脂肪间充质干细胞 wnt3基因 KLOTHO基因 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]