脑出血后继发性脑损伤及其保护作用机制的实验研究
发布时间：2019-03-04 08:46
【摘要】：目的：通过观察大鼠ICH后小胶质细胞（OX-42）ǎNF-κBp65ǎBDNFǎSYPǎCaspase3的动态表达及安脑丸对其表达的干预作用，，探讨安脑丸对ICH后继发性脑损伤的保护作用机制 方法：将Sprague-Dawley(SD)系雄性大鼠190只随机分为正常组ǎ假手术组ǎICH模型组（模型组）ǎ安脑丸干预组(安脑丸组)；后三组组内随机分为术后12hǎ1dǎ2dǎ4dǎ7dǎ10d六个时间点；按Rosenberg法建立大鼠ICH模型，安脑丸组每天上下午各灌胃1mg/ml安脑丸10ml/kg一次分别于术后各时间点对大鼠进行神经功能缺损评分；免疫组化方法检测大鼠脑组织中小胶质细胞ǎNF-κBp65ǎBDNFǎSYNǎCaspase3阳性细胞表达及Western blot法检测大鼠脑组织BDNFǎCaspase3蛋白表达情况 结果：安脑丸组大鼠神经功能缺损评分低于模型组（P0.05）免疫组化法检测结果显示，正常组ǎ假手术组大鼠仅见少量OX42阳性细胞表达，模型组ICH后第12h即可见血肿周围有大量OX42阳性细胞表达，第7-10d表达达到高峰，安脑丸组各时间点OX42阳性细胞表达及活化都无模型组明显（P0.01）；模型组ICH后第12hNF-κBp65表达明显增多，第4d时NF-κBp65表达达到高峰，安脑丸组各时间点NF-κBp65表达均明显少于模型组（P0.05）；模型组ICH后第12h BDNF表达明显增高，第1d时达到高峰，安脑丸组各时间点表达都明显高于模型组（P0.01）；ICH后第1dǎ2dǎ4dǎ7dǎ10d时安脑丸组SYP蛋白表达高于模型组，差异有统计学意义（P0.01）；模型组和安脑丸组ICH后第12h可见Caspase3大量表达，安脑丸组各时间点Caspase3阳性细胞表达明显少于模型组（P0.01） Westernblot检测结果示，模型组在ICH后第12h时BDNF蛋白的表达显著增多，在第1d时达到高峰，安脑丸组各时间点BDNF蛋白表达均明显高于模型组（P0.01）；模型组ICH后第12h时Caspase3蛋白表达明显增高，第1d时达高峰，安脑丸组各时间点Caspase3表达均明显低于模型组（P0.01） 结论：ICH后小胶质细胞即被激活及NF-κBp65ǎBDNFǎSYN与Caspase3高表达，安脑丸可通过显著减少ICH后小胶质细胞活化ǎNF-κBp65及Caspase3高表达，增加BDNF及SYN的表达而对出血脑组织具有一定的保护作用
[Abstract]:Objective: to observe the dynamic expression of microglia (OX-42) NF- kappa Bp65 / BDNF / SYP-Caspase-3 after ICH in rats and the intervention effect of Annao Pill on the expression. To explore the protective mechanism of Annao Pill on secondary brain injury after ICH methods: Sprague-Dawley (SD) male rats were randomly divided into normal group (sham operation group) ICH model group (model group) and Anao pill intervention group (Annao pill group). The rats in the latter three groups were randomly divided into 6 time points, 12 h, 2 d, 4 d, 7 d and 10 d, according to Rosenberg method to establish the rat ICH model. The rats in the Annao pill group were given 1mg/ml Annao Pill 10ml/kg once a day in the morning and afternoon, and the neurological impairment scores were evaluated at each time point after the operation. Immunohistochemical method was used to detect the expression of NF- kappa Bp65 BDNF and SYN Caspase 3 positive cells in rat brain tissue and Western blot method was used to detect the expression of BDNFCaspase3 protein in rat brain tissue. The results showed that the score of neural function defect in the Annao pill group was lower than that in the model. In group A (P0.05), the results of immunohistochemistry showed that, There were only a few OX42 positive cells in the sham-operated rats in the normal group, and a large number of OX42 positive cells were found around the hematoma at 12 h after ICH in the model group, and reached the peak on the 7th-10th day. There was no significant difference in the expression and activation of OX42 positive cells in the Annao pill group at each time point (P0.01). The expression of NF-魏 Bp65 in the model group increased significantly at the 12th hour after ICH, and reached the peak on the 4th day. The expression of NF- 魏 Bp65 in the Annao pill group at each time point was significantly lower than that in the model group (P0.05). The expression of BDNF in the model group was significantly higher than that in the model group at the 12th hour after ICH and reached the peak on the 1st day. The expression of BDNF in the Annao pill group was significantly higher than that in the model group (P0.01). The expression of SYP protein in Annao pill group was significantly higher than that in model group on the 1st day, 2nd day, 4th day, 7th day and 10th day after ICH (P0.01). Significant expression of Caspase3 was observed at the 12th hour after ICH in the model group and Annao pill group. The expression of Caspase3 positive cells in the Annao pill group was significantly lower than that in the model group (P0.01) at each time point. The expression of BDNF protein in the model group increased significantly at the 12th hour after ICH. On the 1st day, the expression of BDNF protein in Annao pill group was significantly higher than that in model group (P0.01). The expression of Caspase3 protein increased significantly at the 12th hour after ICH in the model group, and reached the peak on the 1st day. The expression of Caspase3 in the Annao pill group was significantly lower than that in the model group (P0.01). Conclusion: microglia were activated after ICH and the expression of NF- kappa Bp65 / BDNF syn and Caspase3 was higher than that in the control group (P < 0.01). Annao Pill can significantly reduce the expression of NF- 魏 Bp65 and Caspase3 in microglia activated by ICH, increase the expression of BDNF and SYN, and protect the brain tissue from hemorrhage.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R651.15
本文编号：2434118
[Abstract]:Objective: to observe the dynamic expression of microglia (OX-42) NF- kappa Bp65 / BDNF / SYP-Caspase-3 after ICH in rats and the intervention effect of Annao Pill on the expression. To explore the protective mechanism of Annao Pill on secondary brain injury after ICH methods: Sprague-Dawley (SD) male rats were randomly divided into normal group (sham operation group) ICH model group (model group) and Anao pill intervention group (Annao pill group). The rats in the latter three groups were randomly divided into 6 time points, 12 h, 2 d, 4 d, 7 d and 10 d, according to Rosenberg method to establish the rat ICH model. The rats in the Annao pill group were given 1mg/ml Annao Pill 10ml/kg once a day in the morning and afternoon, and the neurological impairment scores were evaluated at each time point after the operation. Immunohistochemical method was used to detect the expression of NF- kappa Bp65 BDNF and SYN Caspase 3 positive cells in rat brain tissue and Western blot method was used to detect the expression of BDNFCaspase3 protein in rat brain tissue. The results showed that the score of neural function defect in the Annao pill group was lower than that in the model. In group A (P0.05), the results of immunohistochemistry showed that, There were only a few OX42 positive cells in the sham-operated rats in the normal group, and a large number of OX42 positive cells were found around the hematoma at 12 h after ICH in the model group, and reached the peak on the 7th-10th day. There was no significant difference in the expression and activation of OX42 positive cells in the Annao pill group at each time point (P0.01). The expression of NF-魏 Bp65 in the model group increased significantly at the 12th hour after ICH, and reached the peak on the 4th day. The expression of NF- 魏 Bp65 in the Annao pill group at each time point was significantly lower than that in the model group (P0.05). The expression of BDNF in the model group was significantly higher than that in the model group at the 12th hour after ICH and reached the peak on the 1st day. The expression of BDNF in the Annao pill group was significantly higher than that in the model group (P0.01). The expression of SYP protein in Annao pill group was significantly higher than that in model group on the 1st day, 2nd day, 4th day, 7th day and 10th day after ICH (P0.01). Significant expression of Caspase3 was observed at the 12th hour after ICH in the model group and Annao pill group. The expression of Caspase3 positive cells in the Annao pill group was significantly lower than that in the model group (P0.01) at each time point. The expression of BDNF protein in the model group increased significantly at the 12th hour after ICH. On the 1st day, the expression of BDNF protein in Annao pill group was significantly higher than that in model group (P0.01). The expression of Caspase3 protein increased significantly at the 12th hour after ICH in the model group, and reached the peak on the 1st day. The expression of Caspase3 in the Annao pill group was significantly lower than that in the model group (P0.01). Conclusion: microglia were activated after ICH and the expression of NF- kappa Bp65 / BDNF syn and Caspase3 was higher than that in the control group (P < 0.01). Annao Pill can significantly reduce the expression of NF- 魏 Bp65 and Caspase3 in microglia activated by ICH, increase the expression of BDNF and SYN, and protect the brain tissue from hemorrhage.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R651.15
【参考文献】
相关期刊论文 前1条
1 李光勤,董为伟;大鼠局灶性脑缺血后白细胞浸润及强力霉素干预效果的观察[J];中华医学杂志;1998年11期
本文编号：2434118
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