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目的 探讨缺血再灌流脑区半胱氨酸蛋白酶 3(caspase 3)活性变化与神经元凋亡的关系。方法 采用WesternBlotting、原位细胞凋亡检测和免疫组化染色等方法,观察大鼠大脑中动脉栓塞 2h后再灌流 1、6、12、24h时,颞顶叶皮层缺血脑区caspase 3表达和活化、多 (ADP 核糖 )聚合酶(PARP)表达和切割灭活及神经元凋亡程度的变化。结果 再灌流 1、6、12及 24h组,缺血脑区的caspase 3前体及其 12 000切割片段含量的相对灰度值分别为 16 72±2 96、28 36±3 51、51 15±3 10、76 14±3 45及 8 17±2 31、15 36±1 39、31 23±5 43、58 95±6 28;PARP及其 24 000切割片段含量的相对灰度值分别为 12 63±3 02、22 65±4 38、30 81±3 16、67 49±8 59及 6 02±0 73、12 86±2 30、20 76±3 16、27 36±2 63;PARP阳性神经元及凋亡神经元的密度 (细胞数 /0 1mm2 )分别为 68 00±6 67、91 40±9 56、112 20±11 78、127 00±11 51及 83 31±7 53、100 70±6 16、121 53±7 21、197 36±11 78。以上 6种指标的变化彼此间均呈正相关 (r= 0 805 ~0 942, P<0 01 );除 6h与 12h组间PARP含量的差异及 6h与 12h组间和 12h与 24h组间PARP阳性神经元密度的差异无统计学意义外,上述 6种指标的其余组间差异均有统计学意义 (P<0 0
Objective To investigate the relationship between the changes of caspase 3 activity and neuronal apoptosis during ischemia-reperfusion brain damage. Methods Western blotting, in situ cell apoptosis assay and immunohistochemical staining were used to observe the expression of caspase 3 in the ischemic cortex of temporal parietal cortex after reperfusion for 1, 2, 6, 12 and 24 hours after embolization of middle cerebral artery Activation, poly (ADP ribose) polymerase (PARP) expression and cleavage inactivation and neuronal apoptosis. Results The gray value of caspase 3 precursor and its 12 000 cleaved segment in the ischemic brain regions were 16 72 ± 2 96,28 36 ± 3 51 and 51 15 ± 3 10,76 14 ± 3 45 and 8 17 ± 2 31,15 36 ± 1 39,31 23 ± 5 43,58 95 ± 6 28; The relative gray values of PARP and its 24 000 cleavage fragments were 12 63 ± 3 02,22 65 ± 4 38,30 81 ± 3 16,67 49 ± 8 59 and 6 02 ± 0 73,12 86 ± 2 30,20 76 ± 3 16,27 36 ± 2 63 PARP positive neurons And the density of apoptotic neurons (number of cells / 0 1mm2) were 68 00 ± 6 67,91 40 ± 9 56,112 20 ± 11 78,127 00 ± 11 51 and 83 31 ± 7 53,100 70 ± 6 16,121 53 ± 7 21,197 36 ± 11 78. The changes of the above 6 indexes were positively correlated with each other (r = 0 805 ~ 0 942, P <0.01). The differences of PARP content between 6h and 12h groups and PARP positive between 6h and 12h groups and 12h and 24h groups Neuronal density differences were not statistically significant, the remaining six groups of six indicators were statistically significant differences (P <0 0