丁咯地尔对局灶性脑缺血大鼠海马CA1区Bcl-2、Bax及Caspase-3表达的影响

Effects of buflomedil on expressions of Bcl-2,Bax,and Caspase-3 protein in hippocampus CAl area of rats after focal cerebral ischemia

目的观察丁咯地尔对局灶性脑缺血大鼠海马CA1区Bcl-2、Bax及Caspase-3蛋白表达的影响。方法健康雄性SD大鼠随机分为:假手术组,模型组和丁咯地尔低、中、高剂量组,每组6只。线栓法制作大鼠大脑中动脉缺血模型。丁咯地尔各剂量组在造模前7d每日腹腔注射丁咯地尔1次,剂量分别为5、10和20 mg·kg^(-1),并于术前1h再次腹腔注射1次。假手术组予丁咯地尔10 mg·kg^(-1),模型组予以等体积生理盐水代替。免疫组化法观察海马CA1区Bcl-2、Bax和Caspase-3的表达情况。结果光镜下可见,与假手术组比较,模型组大鼠海马CA1区细胞出现核固缩,细胞浆浓缩深染,Bcl-2、Bax及Caspase-3蛋白表达显著增多。与模型组比较,丁咯地尔各剂量组大鼠海马CA1区Bcl-2蛋白表达增多,Bax及Caspase-3蛋白表达减少。随着丁咯地尔剂量的增加,Bcl-2的阳性表达率逐渐增加,Bax和Caspase-3的阳性表达率逐渐降低,Bcl-2/bax比值逐渐增加(p<0-05或P<0.01)。结论丁咯地尔可抑制局灶性脑缺血大鼠海马CA1区的Bax和Caspase-3表达,促进Bcl-2表达,降低脑损伤。

AIM To observe the effects of buflomedil on expressions of Bcl-2, Bax, and Caspase-3 protein in hippocampus CA1 area of rats after focal cerebral ischemia. METHODS Thirty healthy male SD rats were randomized into five groups （n = 6 in each group） ： sham-operation group, model group, low-dose, middle-dose, and high-dose buflomedil group. Rats in the different dosage buflomedil groups were administered with buflomedil 5, 10, and 20 mg·kg^-1 for 7 d （ip, qd） and 1 h before model construction, respectively. Rats in the sham-operation group were given buflomedil 10 mg·kg^-1, volume of normal saline. Middle cerebral artery occlusion model rats in the model group were received an equal was prepared with Longa＇ s method. Expressions of Bcl-2, Bax, and Caspase-3 were detected by immunohistochemical method. RESULTS Compared with the sham-operation group, karyopyknosis, condensed and deeply stained cytoplasm were observed in the model group, and the expressions of Bcl-2, Bax, and Caspase-3 protein increased obviously. Compared with the model group, the expression of Bcl-2 protein increased in the different dosage buflomedil groups, while the expressions of Bax and Caspase-3 protein decreased. With the increasing of buflomedil＇ s dosage, the positive expression rate of Bcl-2 increased, Bax and Caspase-3 decreased, while the value of Bcl-2/Bax increased （P 〈 0.05 或 P 〈 0.01）. CONCLUSION Buflomedil inhibits the expressions of Bax and Caspase-3 in hippocampus CA1 area of rats after focal cerebral ischemia, promotes the expression of Bcl-2 so as to protect brain tissues from cerebral ischemia.