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目的研究胱氨酸(Cystamine)对全脑缺血再灌注损伤后大鼠海马组织型转谷氨酰胺酶(tTG)表达的影响。方法用四血管阻断法制作大鼠全脑缺血再灌注模型。将SD大鼠随机分为假手术组(n=8),全脑缺血组(n=48)及全脑缺血治疗组(n=48),全脑缺血组和全脑缺血治疗组按照缺血再灌注时间点的不同再将大鼠随机分为6h、12 h、1 d、3 d、5 d、7 d六个亚组,每个亚组8只。全脑缺血治疗组大鼠给予Cystamine腹腔注射(0.15 mg/g),全脑缺血组和假手术组大鼠给予生理盐水腹腔注射。TUNEL染色法观察细胞凋亡水平的变化,免疫组织化学方法分析再灌注后不同时间点海马CA1区tTG表达水平的变化。另取52只大鼠随机分为假手术组(n=4)、全脑缺血组(n=24)及全脑缺血治疗组(n=24),全脑缺血组和全脑缺血治疗组按照不同时间点每个亚组4只,免疫印迹方法测定tTG的表达。结果缺血组再灌注24 h后TUNEL阳性细胞明显增加,1、3、5、7 d亚组与假手术组差异有统计学意义(P<0.05);治疗组与缺血组相比,在1、3、5、7 d相应时间点亚组TUNEL阳性细胞数减少(P<0.05)。治疗组海马CA1区tTG平均阳性细胞数于全脑缺血后12 h开始下降,3、5、7 d亚组均明显低于缺血组(P<0.05)。治疗组海马tTG蛋白水平于缺血再灌注后1 d开始降低,3、5和7 d亚组显著低于缺血组(P<0.05)。结论 Cystamine可以降低大鼠全脑缺血再灌注后海马CA1区tTG的表达,从而对神经元细胞起到一定的保护作用。
Objective To investigate the effect of cystamine on the expression of tissue transglutaminase (tTG) in rat hippocampus after global cerebral ischemia-reperfusion injury. Methods A rat model of global cerebral ischemia-reperfusion was established by four-vessel occlusion method. The SD rats were randomly divided into sham operation group (n = 8), global cerebral ischemia group (n = 48) and global cerebral ischemia group (n = 48) The rats were randomly divided into 6 subgroups: 6 h, 12 h, 1 d, 3 d, 5 d and 7 d according to the different time points of ischemia reperfusion. Rats in the global cerebral ischemia group were given intraperitoneal injection of Cystamine (0.15 mg / g). Rats in the global cerebral ischemia group and the sham operation group were injected with saline intraperitoneally. TUNEL staining was used to observe the change of apoptosis. The expression of tTG in hippocampal CA1 region was detected by immunohistochemistry at different time points after reperfusion. Another 52 rats were randomly divided into sham operation group (n = 4), global cerebral ischemia group (n = 24) and global cerebral ischemia group (n = 24) Blood treatment group according to different time points in each subgroup of 4, Western blot method to determine the expression of tTG. Results The TUNEL positive cells in the ischemic group were significantly increased 24 h after reperfusion, the difference was statistically significant between the 1, 3, 5, and 7 d subgroups and the sham operation group (P <0.05). Compared with the ischemic group, At 1, 3, 5 and 7 d, the number of TUNEL-positive cells in the subgroup decreased (P <0.05). The average number of tTG positive cells in CA1 area of hippocampus in treatment group began to decrease 12 h after global cerebral ischemia, and was significantly lower in the 3rd, 5th and 7th d subgroups than in the ischemic group (P <0.05). The level of tTG protein in the hippocampus in the treatment group began to decrease at 1 day after ischemia-reperfusion, and was significantly lower in the 3, 5 and 7-day subgroups than that in the ischemia group (P <0.05). Conclusion Cystamine can reduce the expression of tTG in hippocampal CA1 region after global cerebral ischemia and reperfusion in rats and thus play a protective role on neurons.