落新妇甙对心脏移植效应性T细胞磷脂酰肌醇信号通路的影响  

Effect of astilbin on inhibition of phosphatidy linositol 3-kinase-protein kinase B（PI-3K-PKB）in alioreactive T cells of mouse heart transplantation model with acute rejection

作　　者：高思海[1] 李平[1] 陈涛[1] 徐沁孜[1] 宋定伟[1] 刘力刚[1] 潘铁成[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院胸心外科,武汉430030

出　　处：《中华胸心血管外科杂志》2007年第1期47-49,共3页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：本课题受国家自然学基金资助（30300323.30500656）

摘　　要：目的探讨落新妇甙对心脏移植效应性T细胞磷脂酰肌醇-3-激酶／蛋白激酶B（PI-3K／PKB）信号传导通路的影响。方法分离BALB／C小鼠心肌细胞（2×10s／m1）和C57BL／6小鼠的脾细胞（1×10^6／ml），前者作刺激细胞，后者作反应细胞，进行混合细胞培养，建立小鼠心脏移植急性排斥反应体外模型。实验分4组：对照组，心肌细胞与脾细胞混合培养；3个实验组在对照组基础上，落新妇甙组加入落新妇甙（15μg/ml）；落新妇甙加P13K激动剂组加入落新妇甙（15μg/ml）和P13K激动剂IL-8（20μg/ml）；落新妇甙加P13K抑制剂组加入落新妇甙（15μg／ml）和P13K抑制剂LY294002（20μmol／L）。TUNEL法检测T淋巴细胞凋亡情况。Western blot法检测T细胞P13K和Bcl-2蛋白表达情况。RT-PCR法检测T细胞PKB mRNA表达情况。结果落新妇甙组T细胞凋亡指数明显高于对照组[（74．2±11．7）％对（34．2±9．6）％，P〈0．01]，P13K、PKB、Bcl-2表达显著低于对照组（P〈0．01）。落新妇甙加P13K抑制剂组T细胞凋亡指数显著高于落新妇甙组，P13K、PKB、Bcl-2显著低于落新妇甙组（P〈0．01）。落新妇甙加P13K激动剂组T细胞凋亡指数和Bcl-2表达与对照组的差异无统计学意义（P〉0．05）。结论落新妇甙诱导心脏移植效应性T细胞凋亡与其抑制PI-3K／PKB信号通路有关。Objective To investigate the effect of astilbin on inhibition of phosphatidy linositol 3-kinase-protein kinase B （PI-3K-PKB） in alloreacfive T cells of mouse heart transplantation model with acute rejection. Methods Cardlomyocytes of BALB/C mouse and spleen cells of C57BL/6 mouse were separated , the cardiomyocytes （2 × 10^5/ml）as stimulates and spleen cells （1× 10^6/ml） as responsers were mixed cultured. A model of mouse heart transplantation with acute rejection in vitro was established. There were four study groups. Control group, mixed culture of the cardiomyocytes and spleen cells; Astilbin group, mixed culture of the cardiomyocytes and spleen cells and astilbin（ 15 μg/ml ）. Astilbin ＋ IL-8 group, mixed culture of the cardiomyocytes and spleen cells and aatilbin（15 μg/ml ） and IL-8 （20μg/rnl）. Astilbin ＋ LY294002 group, mixed culture of the cardiomyocytes and spleen cells and aatilbin （15μg/ml） and LY294002 （20 μmol/L）. Apoptosis of T cedis were observed by TUNEL. The expression of P13K and Bcl- 2 in alloreactive T cells were measured by Western blot. The expression of PKB in alloreactive T cells was meaasured by RT-PCR. Results Apoptosis indexes in alloreactive T cells in Astilbin group were found significately higher than those of the control group [ （74.2 ＋±11.7） % vs. （ 34.2 ± 9.6 ） %, P 〈 0.01 ]. The expression of P13K, PKB and Bcl-2 in Astilbin group were significantly lower than control group （ P 〈 0.01 ）. The expression of P13K, PKB and Bcl-2 in Astilbin group were significantly higher than control group （ P 〈 0.01 ）. There were no significant difference between control group and Astilbin ＋ IL-8 group （ P 〉 0.05）. Conclusion The apoptosis of alloreactive T cells of heart transplantation induced by astilbin maybe partially related to its inhibition on expression expression of phosphatidy linositol 3-kinase-protein kinase B （PI-3K-PKB）.

关 键 词：心脏移植 移植物排斥 磷脂酰肌醇3-激酶 蛋白激酶类 落新妇甙 

分 类 号：R686[医药卫生—骨科学]