Yang Xiaoyan, Zhang Li, Jiang Shiqin, Gong Peili, Zeng Fandian
Department of Pharmacology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhongguo Zhong Yao Za Zhi. 2009 Jan;34(1):78-83.
To investigate the effect of dauricine on the apoptosis of neuronal cells and the expression of apoptosis-related proteins in the brain penumbra of rats induced by transient focal cerebral ischemia-reperfusion injury.
Male SD rats were randomly divided into five groups: sham group (Sham), model group (Model), and Dauricine groups of low, middle and high doses. To make the transient focal cerebral ischemia-reperfusion injury model, the middle cerebral artery on the right side of rat was occluded by inserting a nylon suture through the internal carotid artery for 1 h, followed by reperfusion for 24 h after withdrawing the suture. Dauricine groups, different doses of Dauricine (2.5, 5, 10 mg x kg(-1) as low, middle and high dose respectively) were administered intraperitoneally at the beginning of the cerebral ischemia, and at 11 h and 23 h after reperfusion. At the same time, Sham group and Model group was administered saline as controls. Brain samples of rats were treated with paraformaldehyde perfusion fixation 24 h after blood reperfusion and then collected for making pathological sections. Apoptotic changes of neuronal cells in the brain penumbra of rat were evaluated in situ by terminal deoxyribonucleotidyl transferasemediated dUTP-digoxigenin nick end-labelling (TUNEL). Cytochrome C (Cyt-C) release and the expression of caspase -3 and caspase -9 proteins of the ischemic-reperfusion brain tissue were determined by immunohistochemistry assay.
TUNEL-positive cells in groups of middle and high doses of dauricine (18.9 +/- 2.02 and 15.9 +/- 2.9 cells/mm2 respectively) decreased significantly compared with model group (25.5 +/- 3.3 cells/mm2, P<0.05). Cyt-C release and the expression of caspase-3 and caspase-9 proteins in groups of middle and high doses of dauricine were also inhibited compared with Model group (P<0.01).
The mechanism of the neuroprotective effect of dauricine after cerebral ischemia-reperfusion injury may parly, related with an inhibition of neuronal cells apoptosis in the penumbra.
探讨蝙蝠葛碱对短暂性局灶性脑缺血再灌注损伤大鼠脑半暗带神经细胞凋亡及凋亡相关蛋白表达的影响。
雄性SD大鼠随机分为五组:假手术组(Sham)、模型组(Model)、低、中、高剂量蝙蝠葛碱组。采用线栓法插入尼龙线阻塞大鼠右侧大脑中动脉1小时制备短暂性局灶性脑缺血再灌注损伤模型,拔出线栓后再灌注24小时。蝙蝠葛碱组在脑缺血开始时及再灌注后11小时和23小时分别腹腔注射不同剂量蝙蝠葛碱(低、中、高剂量分别为2.5、5、10mg·kg-1)。同时,假手术组和模型组腹腔注射生理盐水作为对照。再灌注24小时后用多聚甲醛灌注固定大鼠脑标本,然后取材制作病理切片。采用末端脱氧核苷酸转移酶介导的dUTP-地高辛缺口末端标记法(TUNEL)原位评价大鼠脑半暗带神经细胞的凋亡变化。采用免疫组织化学法检测缺血再灌注脑组织细胞色素C(Cyt-C)释放及半胱天冬酶-3和半胱天冬酶-9蛋白表达。
中、高剂量蝙蝠葛碱组TUNEL阳性细胞数(分别为18.9±2.02和15.9±2.9个/mm2)较模型组(25.5±3.3个/mm2,P<0.05)明显减少。中、高剂量蝙蝠葛碱组Cyt-C释放及半胱天冬酶-3和半胱天冬酶-9蛋白表达较模型组也受到抑制(P<0.01)。
蝙蝠葛碱脑缺血再灌注损伤后神经保护作用机制可能部分与抑制半暗带神经细胞凋亡有关。