海藻糖对低温储存皮肤β-actin的mRNA转录水平及其蛋白表达的影响  

Effect of trehalose on the protein synthesis and mRNA transcription of β-actin after cryopreserved human skins

作　　者：贾晓明[1] 蔡宏[1] 马彩虹[1] 吴志谷[1] 姚咏明[1] 杨毅[1] 李绍然[1] 陆江阳[1] 

机构地区：[1]解放军总医院第304临床部全军烧伤研究所,北京100037

出　　处：《军医进修学院学报》2006年第5期331-333,共3页Academic Journal of Pla Postgraduate Medical School

基　　金：国家自然科学基金项目(30070298)

摘　　要：目的:观察新型低温保护剂(cryoprotectant,CPA)海藻糖(trehalose)对皮肤骨架蛋白β-肌动蛋白(β-actin)表达及mRNA转录水平的调节作用,以了解在传统低温保护剂中添加海藻糖后是否优于单纯传统低温保护剂的作用。方法:实验将新鲜成人皮肤分为4组,分别经海藻糖-二甲基亚砜(T-D)、二甲基亚砜-丙二醇(D-P)、二甲基亚砜-去血清角质细胞培养液(D-K)、DMEM作为低温保护剂保存,-196℃液氮冻存7 d、14 d复温,设新鲜皮肤为对照组。采用免疫组织化学染色及W estern b lot方法观察β-actin蛋白表达、采用RT-PCR方法对不同低温保护剂保存后皮肤的β-actin基因水平进行深入的研究。结果:T-D组β-actin蛋白表达及基因转录与新鲜组相似(P>0.05),D-P作用其次,其余两组与新鲜组对比均降低明显(P<0.05)。结论:传统低温保护剂中添加海藻糖后对低温储存皮肤β-actin表达的保护作用优于单纯传统低温保护剂。Objective ：To observe the effect of a new cryoprotectant, trehalose, on the protein synthesis and mRNA transcription of β-actin so as to ascertain whether there exists difference between the new method and the traditional methods. Methods： The skins to be cryopreserved were first treated with DMSO-Propyleneglycol, DMSO-KSFM, DMEM and trehalose- DMSO, respectively, to be compared with fresh skin. The histological structure of skin of different groups was observed and analyzed by immunohistochemistry. Furthermore, we investigated the influence of trehalose on cytoskeleton at gene level with RT- PCR. The experiments were carried out 7 days and 14 days after cryopreservation. Results：The histological results showed that the content of β-actin could be well-protected by trehalose. The results of RT-PCR also revealed that skin cryopreserved with trehalose-DMSO was as good as fresh skin, and better than other groups. Conclusion.The results suggest that the traditional cryoprotectant combining with trehalose is better than the traditional cryoprotectant.

关 键 词：海藻糖 低温保存 皮肤 低温保护剂 

分 类 号：R329.499[医药卫生—人体解剖和组织胚胎学]