大鼠脑出血后血肿周围组织活化凋亡蛋白酶3表达与大承气汤的干预

Effect of dachengqi decoction on the expression of activated Caspase-3 around hematoma in rats with intracerebral hemorrhage

目的:观察大鼠脑出血后血肿周围神经元活化凋亡蛋白酶3的表达和血肿变化,及大承气汤的干预作用。方法:实验于2003-10/2004-03在中南大学湘雅医院中西医结合研究所完成。健康SD大鼠65只随机分成4组,分别为正常组(5只),假手术组(20只),模型组(20只),大承气汤组(20只)。除正常组外,其余3组设4个时间点:6h,1,3,5d。每个时间点5只。采用免疫组化方法检测大鼠脑内血肿周围活化凋亡蛋白酶3的表达,用直尺测量不同时间点大鼠脑内血肿的最大直径。结果:65只大鼠进入结果分析。①活化凋亡蛋白酶3阳性细胞计数:正常组未见阳性细胞表达。造模后模型组血肿周围于6h出现阳性表达细胞,1d阳性细胞计数达高峰,3d出现轻度下降,5d明显下降[模型组:6h(24.600±2.338)个,1d(115.900±2.215)个,3d(91.940±3.177)个,5d(67.900±2.261)个]。与假手术组比较差异明显[假手术组:6h(3.120±2.235)个,1d(1.360±2.097)个,P<0.01]。大承气汤组与同时间点模型组比较:6h计数无明显差异,1,3,5d均存在明显差异[大承气汤组:1d(114.440±3.239)个,3d(84.420±3.648)个,5d(51.460±3.038)个,P<0.05或P<0.01]。②血肿体积变化情况:造模后血肿于1d达到最大,3d有所变小,5d变小明显;大承气汤组在5d缩小程度与模型组比较有明显差异[大承气汤组5d(2.000±0.187)mm,模型组5d(2.480±0.148)mm,(t=4.496,P<0.01)]。但在6h,1d,3d与模型组比较血肿缩小程度差别无显著性意义。结论:大鼠脑出血后血肿周围神经元活化凋亡蛋白酶3表达明显上调;大承气汤能减少活化凋亡蛋白酶3的表达,阻止神经元的凋亡,同时也具有一定的促进血肿吸收的作用。

AIM: To observe the expression of activated Caspase3 around hematoma in rats with intracerebral hemorrhage,the changes of hematoma size and the effects of dachengqi decoction. METHODS: The experiment was finished at the Institute of Combined Traditional Chinese and Western Medicine of Xiangya Hospital,Central South University from October 2003 to March 2004.A total of 65 healthy SD rats were divided randomly into 4 groups: normal group(n=5),sham operation group(n=20),model group(n=20) and dachengqi decoction group(n=20). Rats expect in normal group were subdivided into 6hour,1, 3and 5day groups,with 5 rats in each group.Immunohistochemical method was used to detect the expression of activated Caspase3 around hematoma in rats with intracerebral hemorrhage.The diameter of hematoma in the brain at different time was measured with a ruler. RESULTS: All 65 rats were involved in the result analysis.①The expression of positive activated Caspase3 did not show in normal group;however,in the model group,it appeared around hematoma at hour 6 after intracerebral hemorrhage(24.600±2.338),reached to the peak at day 1(115.900±2.215),decreased slightly at day 3 (91.940±3.177),decreased significantly at day 5 (67.900±2.261),and there was significant difference between the model group and the sham operation group[hour 6: 3.120±2.235;day 1:1.360±2.097,P< 0.01].There was significant difference in the positive expression of activated Caspase3 at days 1,3 and 5 except at hour 6 between dachengqi decoction group and model group[1 day: 114.440±3.239,3 day: 84.420±3.648,5 day: 51.460±3.038,P< 0.05].②The size of hematoma increased to the biggest at day 1,decreased slightly at day 3,decreased significantly at day 5 after intracerebral hemorrhage,and there was significant difference in size of hematoma at day 5 between dachengqi decoction group and model group[(2.000±0.187),(2.480±0.148) mm;t=4.496,P< 0.01],but there was no significant difference in the size of hematoma at hour 6,days 1 and 3. CONCLUSION: The expression of activated Caspase3 increases obviously in the neurons around the hematoma after intracerebral hemorrhage.Dachengqi decoction can markedly decrease the expression of activated Caspase3,inhibit neuronal apoptosis,and accelerate the absorbing of the hematoma.