摘要
目的探讨阿加曲班对急性脑梗死大鼠脑组织细胞凋亡数量、caspase-3、Bcl-2和Bax表达的影响。方法建立SD大鼠大脑中动脉缺血(MCAO)模型,将大鼠分为假手术组(Sham组,不插入MCAO拴线)、脑中动脉缺血模型组(MCAO组)和阿加曲班处理组(Argatroban组)。分别选取0、6和12 h共3个时间窗的样本进行检测,TUNEL法细胞凋亡检测试剂盒检测大鼠脑部神经细胞凋亡,Western blot和ELISA法检测大鼠脑部caspase-3、Bax和Bcl-2水平。多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验。结果TUNEL法结果显示,与MCAO组比较,Sham组和Argatroban组在0、6、12 h的细胞凋亡指数均降低[0 h(14.91±4.11)﹪比(2.13±0.58)﹪、(4.22±1.01)﹪;6 h(24.75±3.88)﹪比(3.19±0.28)﹪、(12.14±1.90)﹪;12 h(48.22±2.69)﹪比(5.75±1.21)﹪、(28.45±7.84)﹪,P均<0.05]。ELISA结果显示:与Sham组比较,MCAO组0、6、12 h的大鼠脑组织细胞caspase-3、Bax表达均升高,而Bcl-2表达在6、12 h时升高(P均<0.05);与MCAO组比较,Argatroban组的脑组织细胞caspase-3、Bax表达均降低,而Bcl-2表达在6、12 h时升高[caspase-3表达:0 h(2.32±0.12)比(1.25±0.06)、6 h(5.91±0.60)比(3.26±0.22)、12 h(7.31±0.27)比(6.42±0.18)ng/ml;Bax表达:0 h(0.82±0.08)比(0.45±0.03)、6 h(1.00±0.05)比(0.66±0.04)、12 h(1.56±0.11)比(1.09±0.07)ng/ml;Bcl-2表达:6 h(1.07±0.08)比(1.56±0.05)ng/ml,12 h(0.67±0.08)比(1.45±0.06)ng/ml,P均<0.01]。Western blot结果显示,大鼠脑组织细胞caspase-3、Bax蛋白表达趋势与ELISA结果一致;而Bcl-2蛋白表达在Argatroban组高于Sham组和MCAO组,差异有统计学意义(P均<0.01)。结论阿加曲班可通过减少急性脑梗死诱导的大鼠脑组织细胞凋亡、抑制caspase-3和Bax表达,并上调Bcl-2水平而产生神经保护作用。
Objective To investigate the effect of argatroban on neuronal apoptosis,and the expression of caspase-3,Bcl-2 as well as Bax in rats with acute cerebral infarction.Methods The rat model of focal cerebral ischemia was established by middle cerebral artery occlusion(MCAO).SD rats were divided into Sham group,MCAO model group and Argatroban treatment group,and 3 time windows were set(0,6,12 h).The samples of 3 time windows at 0,6,12 h were selected for detection,respectively.TUNEL apoptosis detection kit was used to detect neuronal apoptosis in the rat brain;Western blot and ELISA were used to detect the expression of caspase-3,Bcl-2 and Bax in the rat brain;ANOVA was used for comparison among groups,and LSD-t test was used for comparison of two groups.Results The results of TUNEL showed that the apoptosis index of the sham group and Argatroban group all decreased at 0,6,12 h when compared with MCAO group[0 h(14.91±4.11)﹪vs(2.13±0.58)﹪,(4.22±1.01)﹪;6 h(24.75±3.88)﹪vs(3.19±0.28)﹪,(12.14±1.90)﹪;12 h(48.22±2.69)﹪vs(5.75±1.21)﹪,(28.45±7.84)﹪,P<0.05].The results of ELISA showed that the expression of caspase-3 and Bax in rat brain neuron in MCAO group increased at 0,6 and 12 h compared with that in the sham group,while the expression of Bcl-2 increased at 6 and 12 hours(P<0.05).And compared with MCAO group,the expression of caspase-3 and Bax in rat brain neuron in Argatroban group decreased at 0,6 and 12 h,while the expression of Bcl-2 increased at 6 and 12 hours[the expression of caspase-3:0 h(2.32±0.12)vs(1.25±0.06)、6 h(5.91±0.60)vs(3.26±0.22)、12 h(7.31±0.27)vs(6.42±0.18)ng/ml;the expression of Bax:0 h(0.82±0.08)vs(0.45±0.03)、6 h(1.00±0.05)vs(0.66±0.04)、12 h(1.56±0.11)vs(1.09±0.07)ng/ml;the expression of Bcl-2:6 h(1.07±0.08)vs(1.56±0.05)ng/ml,12 h(0.67±0.08)vs(1.45±0.06)ng/ml,P<0.01].The protein expression trends of caspase-3 and Bax in rat brain neuron among 3 groups were consistent with those of ELISA,while the expression of Bcl-2 protein in Argatroban group was high
作者
刘猛
刘娟
朱月敏
袁亚松
韩建鹏
王姣姣
卢双动
Liu Meng;Liu Juan;Zhu Yuemin;Yuan Yasong;Han Jianpeng;Wang Jiaojiao;Lu Shuangdong(Department of Internal Neurology,Zhuozhou City Hospital,Zhuozhou 072750,China;Department of Emergency Internal Medicine,Baoding Second Central Hospital,Baoding 072750,China)
出处
《中华细胞与干细胞杂志（电子版）》
2020年第1期37-43,共7页
Chinese Journal of Cell and Stem Cell（Electronic Edition）
基金
2019年保定市科学技术研究与发展指导计划(第一批)项目(18zf153)。
