含川芎嗪的玻璃化保存液对雪旺细胞凋亡及调控基因表达影响的初步实验研究  

Preliminary experimental study of the effects of vitreous cryopreservation with tetramethylpyrazine on schwann’s cell apoptosis and the expression of regulating gens

作　　者：刘建文[1] 蒋电明[1] 陈增刚[1] 袁新[1] 欧云生[1] 

机构地区：[1]重庆医科大学附属第一医院骨科,重庆400016

出　　处：《重庆医科大学学报》2009年第3期289-292,共4页Journal of Chongqing Medical University

摘　　要：目的:探讨在不同温度下含川芎嗪的玻璃化保存液对大鼠雪旺细胞凋亡及调控基因表达的影响。方法:24只SD大鼠,切取双侧坐骨神经(共48条),将其随机分成2组(每组24条):实验组(含川芎嗪)和对照组(不含川芎嗪)。实验组分为A、B、C、D亚组,对照组分为A’、B’、C’、D’亚组(每亚组6条),分别在不同的温度下保存3周(A、A’亚组:-4℃保存;B、B’亚组:-20℃保存;C、C’亚组:-80℃保存;D、D’亚组:-196℃保存)。3周后取神经进行检测:①HE染色光镜检查;②免疫组化法检测Bcl-2、Bax;③TUNEL法检测细胞凋亡。结果:除D、D’亚组外,实验组各亚组神经雪旺细胞凋亡数量明显少于对照组相应的各亚组,实验组各亚组Bcl-2的阳性表达率均明显高于对照组相应的各亚组,实验组各亚组Bax阳性表达率均明显低于对照组相应的各亚组,差异均有显著性(P<0.05);实验组中B亚组雪旺细胞凋亡情况和Bcl-2与Bax的表达情况与C、D亚组接近,比较无显著性差异(P>0.05)。结论:含川芎嗪的玻璃化保存液能够在-20℃条件下,通过调节Bcl-2和Bax的表达而有效抑制雪旺细胞的凋亡。Objective：To investigate the effects of vitreous cryopreservation with tetramethylpyrazine on schwann＇s cell apoptosis and the expression of regulating gens under different temperatures. Methods： 48 sciatic nerves obtained from 24 SD rats were randomly divided into 2 groups （each with 24 nerves）： experimental group （with tetramethy -lpyrazine, 320mg/L ） and control group （no tetramethylpyrazine）. Experimental group included 4 subgroups ：A B C D;and the control group included 4 subgroups： A＇,B＇,C＇, D＇ （each with 6 nerves）. All the corresponding subgroups were cryopreserved 3 weeks under different temperatures（ A A＇ ： -4℃,B B＇ ： -20℃,C C＇ ： -80℃, D D＇： - 196℃） .After 3 weeks, all the nerves were obtained for tests： ①HE staining for light-microscopetest, ② Schwann ＇S cell apoptosis was measured quantitatively by the terminal transferase UTP nick end labeling （TUNEL） method, ③The expression of Bcl-2 and Bax was determined by immunohistochemisty technique. Results： Aside from D D＇ subgroups, the number of Schwann＇ s cell apoptosis in each experimental subgroup was significantly smaller than that in the corresponding control subgroup （P〈0. 05 ）;the expression of Bcl-2 were significantly increased in each experimental subgroup compared with that in the corresponding control subgroup and the expression of Bax decreased significantly（P〈0.05）. In the experimental group, the rate of Schwann＇S cell apoptosis and that of the positive expression of Bcl-2（or Bax） in B subgroup were approximately equal to those in C and D subgroup（P〉0.05 ）. Conclusion： These findings suggest that at -20℃ vitreous cryopreservation with TMP can depress Schwann＇ S cell apoptosis of rat by regulating the expression of Bcl-2 and Bax.

关 键 词：川芎嗪 雪旺细胞 玻璃 化保存液 细胞凋亡 

分 类 号：R318.06[医药卫生—生物医学工程]