落新妇甙对心脏移植排斥反应中活化T细胞蛋白激酶C信号传导通路的影响  

Effect of astilbin on the inhibition of protein kinase C in activated 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：《中华小儿外科杂志》2007年第1期35-38,共4页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金资助项目（30300323）

摘　　要：目的 用小鼠心脏移植急性排斥反应体外模型，探讨落新妇甙对心脏移植排斥反应中活化T细胞蛋白激酶C（protein klnase C，PKC）信号传导通路的影响。方法 分离BALB／C小鼠心肌细胞（2×10^5／ml）和C57BL／6小鼠的脾细胞（1×10^6／ml），前者做刺激细胞，后者做反应细胞，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；落新妇甙组，在对照组基础上加入落新妇甙（15μg／ml）；落新妇甙加PKC激动剂组，在对照组基础上加入落新妇甙（15μg／ml）和PKC激动剂PMA（10nmol／L）;落新妇甙加PKC抑制剂组，在对照组基础上加入落新妇甙（15μg／ml）和PKC抑制剂CHE（10μmol／L）。TUNEL法检测T淋巴细胞凋亡情况。RT-PCR和Western blot法检测T细胞PKC、NF-κB，easpase-3和Bcl-2表达情况。结果 落新妇甙组T细胞凋亡指数和caspase-3明显高于对照组（P〈0．01），PKC、NF-κB和Bcl-2表达显著低于对照组（P〈0．01）。落新妇甙加PKC抑制剂组T细胞凋亡指数和caspase-3显著高于落新妇甙组，PKC、NF—κB和Bcl-2表达显著低于落新妇甙组（P〈0．01）。落新妇甙加PKC激动剂组T细胞凋亡指数，caspase-3、PKC、NF-κB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排斥反应中活化T细胞凋亡与其抑制PKC信号传导通路有关。Objective To investigate the effect of astilbin on the inhibition of protein kinase C （PKC） in activated T cells of mouse heart transplantation model with acute rejection. Methods Cardiomyocytes of BALB/C mouse and spleen cells of C57BL/6 mouse were separated, the cardiomyocytes （2 × 10^5 ml^- 1 ） as stimulates and spleen cells （ 1 × 10^6 ml^-1 ） as responsers were cultured together. The model of mouse heart transplantation with acute rejection in vitro was established in 4 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 ＋ PMA group, mixed culture of the cardiomyoeytes and spleen cells and astilbin （15 μg/ml） and PMA （10 nmol/L） ; and Astilbin ＋ CHE group, mixed culture of the cardiomyocytes and spleen cells and astilbin （15 μg/ml） and CHE （10 μmol/L）. Apoptosis of T cells were visualized by TUNEL. The expressions of PKC, NF-κB, caspase-3 and Bcl-2 in activated T cells were measured by RTPCR and Western blotting. Results Apoptosis indexes and the expression of easpase-3 in activated T cells in Astilbin group were significant higher than those of the control group （P〈0. 01）, while the expressions of PKC, NF-κB and Bcl-2 in Astilbin group were significantly lower than those in control group（P〈0. 01）. There were no significant difference of these expressions between control group and Astilbin ＋ PMA group. Conclusions Apoptosis of activated T cells of heart transplantation induced by Astilbin maybe partially related to its inhibition expression of protein kinase C （PKC）.

关 键 词：心脏移植 蛋白激酶C 

分 类 号：R686[医药卫生—骨科学]