同种瓣论文：不同浓度海藻糖对液氮深低温保存同种带瓣大动脉组织的保护作用及其对ca spase-3表达的影响.docVIP

同种瓣论文：不同浓度海藻糖对液氮深低温保存同种带瓣大动脉组织的保护作用及其对caspase-3表达的影响 【中文摘要】观察海藻糖作为冷冻保护剂冻存同种带瓣大动脉组织,寻求最佳的冷冻保护剂并探讨冷冻保护剂的最佳适宜浓度以及带瓣大动脉组织caspase-3的表达。方法在液氮中冻存时间点分为12个月,15个月,18个月,依据冻存液不同分为5组,分别是：0.1mol/L DMSO(对照组),0.1mol/L海藻糖(实验组1),0.1mol/L海藻糖+0.1mol/L DMSO(实验组2),0.2mol/L海藻糖+0.1mol/L DMSO(实验组3),0.3mol/L海藻糖+0.1mol/L DMSO(实验组4)。免疫组化光镜观察组织细胞凋亡,测定血糖浓度检测组织细胞代谢情况。采用RT-PCR和Western Blot方法分别测定Caspase-3的相对表达量。新鲜组作为阴性对照。结果随着保存时间的延长,同种带瓣大动脉组织和细胞破坏逐渐加重。光镜下观察,新鲜大动脉组织均为弱阳性,各组标本与新鲜组织比较均有明显改变(d=37-43,P0.05)。各组之间除实验组2和实验组3比较无差异外(d=66-72,P0.05)其余各组比较均有差异(d=0-42,P0.05),细胞凋亡最轻的是新鲜组,较好的是实验组2和实验组3,其次依次为实验组4、实验组1,凋亡最严重的是对照组。血糖测定,实验组2和实验组3无统计学意义(P=0.680.05),其余各组标本有显著的统计学意义(F=41-46,P0.05),细胞代谢最旺盛的是新鲜组,较好的是实验组2和实验组3,其次依次为实验组4、实验组1,代谢最弱的是对照组。RT-PCR和Western-Blot,在每个时间点下(P0.05),细胞凋亡最轻的是新鲜组,较好的是实验组2和实验组3,其次依次为实验组4、实验组1,凋亡最严重的是对照组。结论1、海藻糖对液氮保存带瓣大动脉有较好的保护作用;2、0.1mol/L海藻糖+0.1mol/L DMSO和0.2mol/L海藻糖+0.1mol/L DMSO联合应用,其保护效果好于0.3mol/L海藻糖+0.1mol/L DMSO以及单独使用0.1 mol /L DMSO和0.1mol/L海藻糖。3、海藻糖作为冷冻保护剂冻存同种带瓣大动脉组织,最佳的保存方案是0.1mol/L海藻糖+0.1mol/L DMSO和0.2mol/L海藻糖+0.1mol/L DMSO。 【英文摘要】To observe the effect of trehalose as cryoprotectant for preserving aortic valve homograft in liquid nitrogen on their tissues and seek the best protective agents and their best concentrations, otherwise, detect the expression of caspase-3.Methods The aortic valve homograft was divided into 5 groups, namely:0.1mol/L DMSO (control group), O.lmol/L trehalose (experimental group 1), 0.1mol/L trehalose +0.1mol/L DMSO (experimental group 2),0.2mol/L trehalose+0.1mol/L DMSO (experimental group 3),0.3mol/L trehalose+0.1mol/L DMSO (experimental group 4). At the time of 12 months,15 months and 18 months after preserved in liquid nitrogen, the expression of caspase-3 of the aortic valve homograft was measured by the light microscopy RT-PCR and Western Blot. At the same time, the glucose consumption of each group in 24 Hours was detected to represent the cell metabolism. Fresh group was a negative control group.Results The longer of the samples stored, the worse of