摘要
目的通过Bcl-2、Bax、Caspase-3基因的表达探讨在胸骨低温保存中海藻糖的保护机制。方法切取SD大鼠胸骨后立即放入含有新鲜配制4组保存液的冻存管中,Ⅰ组LPD;Ⅱ组LPD+10%DMSO;Ⅲ组LPD+0.15 mol·L^(-1)海藻糖;Ⅳ组LPD+10%DMSO+0.15 mol·L^(-1)海藻糖。程序降温至-80℃后放入液氮中保存。120 d后对胸骨标本进行HE染色和Bcl-2、Bax、Caspase-3蛋白检测(免疫组化)及RT-PCR扩增。结果大鼠胸骨在4种保存液低温保存120 d后,Ⅲ组和Ⅳ组中的胸骨结构完整,骨细胞Bax、Caspase-3基因表述较低,Bcl-2基因表达较高,尤其Ⅳ组保存效果最好,同种异体异位移植后生长良好。结论在胸骨低温保存中,海藻糖可有效促进Bcl-2基因的表达,抑制骨细胞Bax基因的表达及减少Caspase-3基因的激活,从而抑制骨细胞凋亡,实现对骨细胞的保护。海藻糖和DMSO联合使用保护作用更好。
Objective To explore the protective mechanism of trehalose in sternum cryopreservation. Methods The sternums were removed from SD rats and immersed immediately in the freezing medium of LPD solution only （Group Ⅰ ）, containing with 10% DMSO （Group Ⅱ ）, containing with 0. 15 mol · L^-1 trehalose （Group Ⅲ） and containing with 10% DMSO and 0. 15 mol·L^-1 trehalose （Group Ⅳ） respectively. Sterile plastic tubes with them were then sealed, and frozen to -80 ℃ in a programmable freezer. They were stored in liquid nitrogen （-196℃） for 120 days. Then histologic changes of the sternums were examined after thawing in each group. Bcl-2, Caspase-3 and Bax gene expressions were also observed by immunohistochemistry. At last, thawed and transplanted into the abdominal cavities some sternums （SD） after cryopreservation were of Wistar rats. The transplanted sternums were retrieved and assessed histologically. Results Microscopic findings of the sternums in Group Ⅲ and Group Ⅳ showed their structures were intact and Bax gene expression was lower in cartilage after cryopreservation （120 d） compared with other groups, especially in Group Ⅳ. The sternums in Group Ⅲ and Group iV grew well after they were transplanted into cavities of Wistar rats heterotopieally, too. Conclusion In sternum cryopreservation the trehalose can protect the sternum cartilage by incitating the Bcl- 2 gene and inhibiting the Bax,caspase-3 gene expressions of cartilage cells. The concomitant use of trehalose and DMSO has a synergistic effect.
出处
《中国海洋药物》
CAS
CSCD
2010年第5期33-39,共7页
Chinese Journal of Marine Drugs
关键词
海藻糖
低温保存
胸骨移植
基因表达
trehalose
eryopreservation
sternum transplantation
gene expression
