海藻糖对低温储存皮肤组织桥粒芯糖蛋白2表达及皮肤活力影响的实验研究

目的：比较海藻糖与传统低温保护剂丙二醇对皮肤低温保存后桥粒芯糖蛋白2（Dsg2）表达以及皮肤活力的影响,为寻求皮肤组织低温保护剂的优良配方提供实验依据。方法：经患者同意在烧伤后整形术中获取患者新鲜皮肤组织,分为对照组（新鲜皮肤,不作任何保护剂处理）、海藻糖/二甲基亚砜（T/D ）组（皮肤以0.5 mol/L T/D液浸泡）、二甲基亚砜/丙二醇（D/P）组（皮肤以 D/P 液浸泡）,液氮冻存21 d,复温,Dsg2单克隆抗体免疫组化染色并用 Image Pro5.0图像分析处理系统测定皮肤组织 Dsg2表达的光密度值；用皮肤内琥珀酸脱氢酶（SDH）比色法检测皮肤组织 SDH 活性,以此表示皮肤活力。结果：低温保存21 d时,D/P组Dsg2表达（0.247±0.006）显著低于T/D组（0.250±0.009,P＜0.05）与对照组（0.254±0.007,P＜0.05）；对照组 SDH 水平为18.1±0.6,T/D组为17.4±0.9,D/P组为15.7±0.9,D/P 组 SDH 明显低于 T/D组（P＜0.01）。结论：海藻糖对皮肤组织及细胞间桥粒连接的保护作用优于传统低温保护剂。

Experimental study on the effects of trehalose on viability and desmoglein2 expression in cryopreserved human skin

Objective：To compare the effects of trehalose and traditional cryoprotectant,propyleneglycol,on ex-pression of desmoglein2 （Dsg2）and viability of cryopreserved human skin,and to provide the experimental basis for an ideal formula of cryoprotectant. Methods：The skin specimens which were obtained from burn patients un-dergoing plastic surgery with their consent,were divided into three groups：control group （fresh skin,without any protection against low temperature）,trehalose/dimethyl sulfoxide （DMSO）group （T/D group,treated with 0.5 mol/L trehalose/DMSO）,DMSO/propyleneglycol group（D/P group,treated with DMSO/propyleneglycol）. After being preserved for 2 1 days at -1 9 6 ℃,and rewarmed,then the expression of Dsg2 was observed by the im-munohistochemical method,and optical density value was assessed by Image Pro5.0 system. Furthermore,The viability of skin in 3 groups was evaluated by using succinate dehydrogenase（SDH）. Results：After 21-day cryopr-eservation,optical density value of Dsg2 in D/P group （0.247±0.006）was significantly lower than that in T/D group （0.250±0.009）and control group （0.254±0.007）respectively（P〈0.05）.At the same time,activity of SDH in T/D group （17.4±0.9）was significantly higher than that of the D/P group （15.7±0.9,P〈0.01）.Con-clusions：The results suggest that trehalose is a better cryoprotectant than the traditional cryoprotectant for cryo-preservation of desmosome of human skin.