摘要
目的研究PBK-AKT信号转导系统成员丝氨酸一苏氨酸蛋白激酶（serine／threonine kinase，AKT）、半胱氨酸天冬氨酸蛋白酶3（Caspase-3）在小鼠深低温缺血再灌注脑损伤中的作用。方法将60只4周龄C57／BL6小鼠随机平均分为假手术组（sham group）、模型组（model group）和阻断剂组（LYgroup）建立深低温I／R模型，于再灌注24h后处死小鼠取脑，Western-blot检测AKT1的表达，RT-PCR技术检测小鼠脑组织Caspase-3mRNA，Tunel法检测小鼠大脑皮层细胞凋亡。结果与假手术组比较模型组于再灌注24h后p-AKT显著激活（P〈0．05），Caspase-3mRNA表达明显增加[（0．78±0．03）vs．（0．29±0．05），P〈0．05]并且出现明显的病理凋亡[（75．86％±4．68％）VS．（5．06％±2．17％），P〈0．05]；应用PBK特异性阻断剂LY294002后，阻断剂组小鼠p-AKT表达较模型组减少，且Caspase-3mRNA、病理凋亡均显著高于模型组[（0．90±0．02）vs．（0．78±0．03）；（80．92％±3．18％）vs．（75．86％±4．68％），P〈0．05）]。结论在小鼠脑深低温缺血再灌注过程中PI3K-AKT起到一定的保护作用。
Objective To explore the role of AKT-Caspase-3 signaling pathway on mice brain injury model induced by deep hypothermic cerebral ischemia/reperfusion （I/R）. Methods Sixty C57BL6 mice （four week old） were randomly divided into three groups involving sham operation group, model group and LY group. I/R model was created and brain tissue was harvested at 24 h after reperfusion. The expression of AKT protein was detected using Western-blot assay, Caspase-3 mRNA level was investigated using RT-PCR, and TUNEL assay was utilized to evaluate the cerebral apoptotic cells in each group. Results Compared with the sham operation group, increased expression level was found in p-AKT protein （P〈0. 05） and caspase-3 mRNA [（0. 78 ± 0. 03）vs. （0. 29 ± 0. 05）],as well as higher apoptotic rate [（75. 86% ± 4. 68%） vs. （5. 06± 2. 17%）] in model group, lower expres- sion of p-AKT protein and caspase-3 mR.NA, as well as higher apoptotic rate could be found in LY group in contrast to model group [（0. 90 ± 0. 02） vs. （0. 78 ± 0. 03） ; （80. 92% ± 3.18%） vs. （75.86% ± 4. 68%）,P〈0. 05]. Conclusions Our results suggest that AKT-Caspase-3 signaling pathway has protective effect against deep hypothermic cerebral I/R injury.
出处
《中华小儿外科杂志》
CSCD
北大核心
2012年第12期939-941,共3页
Chinese Journal of Pediatric Surgery
