通窍活血汤对大鼠脑出血的防治机制研究
发布时间：2019-05-30 03:50
【摘要】：目的:通过观察通窍活血汤对急性脑出血大鼠神经功能、NSE、BDNF、SOD、MDA、脑水肿的影响,初步探讨通窍活血汤对大鼠脑出血防治的可能机制。方法:1.分组:取体重在(250±20)g的雄性SD大鼠84只,随机分为假手术组、模型组、依达拉奉组(简称“西药组”)、通窍活血汤高剂量组(简称“高剂量组”)、通窍活血汤中剂量组(简称“中剂量组”)、通窍活血汤低剂量组(简称“低剂量组”)每组各14只;2.造模方法:选用鼠尾自体采血注射法造模,假手术组除不注血外其余操作步骤同其他各组;3.给药方法:西药组给予依达拉奉1.5mg/kg/d腹腔注射,高、中、低剂量组分别给予通窍活血汤1.2g/Kg/d、0.6g/Kg/d、0.3g/Kg/d灌胃,假手术组、模型组给予相同剂量生理盐水灌胃,各组连续给药14天后造模;各组大鼠完成造模后,每天一次神经功能缺损评分,选用Zea Longa5分制法进行,选取神经功能缺失评分在1-3分者作为给药对象;大鼠清醒后,假手术组、模型组继续给予生理盐水灌胃,低、中、高剂量组和西药组同造模前给药方法继续给药1周;4.取材:末次灌胃及注射给药后各组大鼠均采用腹腔麻醉法,分离腹主动脉并穿刺采血;采血后立即心脏灌注并断头取脑组织。5.指标检测:Zea Longa5分制法进行神经功能缺失评分,SOD活性的检测按照说明采用黄嘌呤氧化酶法,MDA含量的检测采用硫代巴比妥酸法;ELISA法检测NSE含量;免疫组化法检测BDNF含量,光镜观察脑出血后血肿周围神经细胞形态变化。采用干湿重法检测脑组织含水量。结果:(1)神经功能缺失评分比较:与假手术组比较,其余各组神经功能缺失评分均升高,差异均有统计学意义(P0.05)。与模型组比较,低剂量组神经功能缺失评分有所降低,但差异无统计学意义(P0.05),中、高剂量组、西药组神经功能缺失评分明显降低,差异均有统计学意义(P0.05)。与低剂量组比较,中、高剂量组、西药组神经功能缺失评分降低,差异均有统计学意义(P0.05)。与中剂量组比较,高剂量组、西药组神经功能缺失评分降低,差异有统计学意义(P0.05)。高剂量组与西药组比较,神经功能缺失评分变化不明显,差异无统计学意义(P0.05)。(2)SOD活性比较:与假手术组比较,模型组、低剂量组、中剂量组、高剂量组、西药组SOD活性下降,差异有统计学意义(P0.05)。与模型组比较,低、中、高剂量组及西药组SOD活性有所增加,差异有统计学意义(P0.05)。与低剂量组比较,中、高剂量组及西药组的SOD活性增加,差异有统计学意义(P0.05)。与中剂量组比较,高剂量和西药组SOD活性增加,差异有统计学意义(P0.05)。与西药组比较,高剂量组SOD活性增加,差异有统计学意义(P0.05)。(3)MDA含量比较:与假手术组比较,其余各组MDA含量均增加,差异有统计学意义(P0.05)。与模型组比较,低剂量组MDA含量有所减少,但差异无统计学意义(P0.05),中、高剂量组及西药组MDA含量明显减少,差异有统计学意义(P0.05)。与低剂量组比较,中、高剂量组及西药组MDA含量明显减少,差异有统计学意义(P0.05)。与中剂量组比较,高剂量组和西药组MDA含量减少,差异有统计学意义(P0.05)。与西药组比较,高剂量组MDA含量有所减少,但差异无统计学意义(P0.05)。(4)BDNF含量比较:与假手术组比较,其他各组的BDNF含量均增加,差异有统计学意义(P0.05)。与模型组比较,低剂量组BDNF表达有所增加,但差异无统计学意义(P0.05);中、高剂量组及西药组BDNF表达明显增加,差异有统计学意义(P0.05)。与低剂量组比较,中剂量组BDNF表达有所增加,但差异无统计学意义(P0.05);高剂量组及西药组BDNF表达明显增加,差异有统计学意义(P0.05)。与中剂量组比较,高剂量组及西药组BDNF表达明显增加,差异有统计学意义(P0.05)。高剂量组与西药组比较,BDNF变化不明显,差异无统计学意义(P0.05)。(5)NSE含量比较:与假手术组比较,模型组、低、中剂量组NSE含量均显著增加,差异有统计学意义(P0.05);高剂量组及西药组NSE含量增加不明显,差异无统计学意义(P0.05)。与模型组比较,低剂量组NSE含量变化不明显,差异无统计学意义(P0.05),中、高剂量组及西药组NSE含量降低,差异有统计学意义(P0.05)。与低剂量组比较,中、高剂量组及西药组NSE含量降低,差异有统计学意义(P0.05)。与中剂量组比较,高剂量组、西药组NSE含量降低,差异有统计学意义(P0.05)。高剂量组与西药组比较,NSE含量变化不明显,差异无统计学意义(P0.05)。(6)脑含水量比较:与假手术组比较,模型组及其余各给药组脑组织含水量均增加,差异有统计学意义(P0.05)。与模型组比较,低剂量组、中剂量组、高剂量组及西药组脑组织含水量均减少,差异有统计学意义(P0.05)。与低剂量组比较,中剂量组、高剂量组及西药组脑组织含水量减少,差异有统计学意义(P0.05)。与中剂量组比较,高剂量组及西药组脑组织含水量减少,差异有统计学意义(P0.05)。高剂量组与西药组比较,脑组织含水量变化不明显,差异无统计学意义(P0.05)。(7)光镜结果:光镜下观察,假手术组神经细胞结构完整,胞核、胞浆染色均匀,未见异常改变。模型组可见神经元及细胞间隙发生水肿,组织结构疏松,神经细胞坏死,血肿周围有少量的炎症细胞浸润。低、中、高剂量组可见不同程度的脑组织水肿及炎症细胞浸润,但随着给药剂量的增高,水肿程度及炎症反应逐渐降低。西药组血肿周围神经细胞结构排列相对完整,未见明显的脑水肿及炎症细胞浸润。结论:1.通窍活血汤可改善脑出血大鼠神经功能缺失评分,减轻脑水肿,改善脑出血后血肿周围神经细胞变性坏死及炎症细胞浸润。2.可能的机制:降低MDA含量,提高SOD活性达到清除氧自由基;促进BDNF的表达,降低NSE含量,从而保护神经细胞。3.通窍活血汤在防治大鼠脑出血作用方面具有一定量效关系。
[Abstract]:Objective: To study the effect of Tongqiao Huoxue Decoction on the neurological function, NSE, BDNF, SOD, MDA and brain edema in rats with acute cerebral hemorrhage. Method:1. Group:84 male SD rats weighing (250 to 20) g were randomly divided into sham operation group, model group, Edarone group (short "Western medicine group"), Tongqiao Huoxue decoction group (short "high-dose group"), Tongqiao Huoxue decoction (short "mid-dose group"), Each group of the low-dose group (low-dose group ") of Tongqiao Huoxue Decoction (the low-dose group") was 14;2. Methods: The model was made by using the self-sampling method of the mouse tail, and the other operation steps of the sham-operated group were similar to that of the other groups. The method comprises the following steps of: administration of edaravone 1.5 mg/ kg/ day in the western medicine group into the abdominal cavity for intraperitoneal injection, wherein the high, middle and low dose groups are respectively provided with the Tongqiao Huoxue decoction 1.2 g/ Kg/ day, 0.6 g/ Kg/ day, 0.3 g/ Kg/ day, the sham operation group and the model group are given the same dose of normal saline for oral administration, each group is continuously administered for 14 days to form a model; and after the rats are finished, Each day, the neurological deficit score was scored, the Zea Longa 5 method was used to make the score, and the neurological deficit score was selected as the administration target; after the rats were awake, the sham-operation group and the model group continued to give the normal saline to the gastric administration, the low, medium, The high-dose group and the western medicine group were treated with the pre-model administration method for 1 week;4. Materials: The rats in each group after the last intragastric administration and after injection were treated with intraperitoneal anesthesia, the abdominal aorta was separated and the blood was punctured, and the heart was perfused immediately after blood collection and the brain tissue was decapitated. The results showed that the method of Zea Longa 5 was used to test the loss of nerve function, and the activity of SOD was detected by using the method of the method of using the method of the method of the method of the method of using the thiobarbituric acid, the content of the MDA was detected by the ELISA, the content of the BDNF was detected by the method of immunohistochemistry. The morphological changes of the nerve cells around the hematoma after cerebral hemorrhage were observed by light microscope. The water content of brain tissue was detected by dry and wet method. Results: (1) The score of the neurological deficit score was compared with that of the sham operation group. The scores of the neurological deficit in the other groups were increased and the difference was statistically significant (P0.05). Compared with the model group, the score of neurological deficit in the low-dose group was lower, but the difference was not significant (P0.05). In the middle and high dose group, the neurological deficit score of the western medicine group was significantly decreased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the functional deficit score of the middle and high dose group and the western medicine group was lower, and the difference was statistically significant (P0.05). Compared with the middle-dose group, the functional deficit score of the high-dose group and the western medicine group was lower, and the difference was statistically significant (P0.05). There was no significant difference in the score of neurological deficit in the high-dose group and the western medicine group (P0.05). (2) Compared with sham operation group, the activity of SOD in the group, low-dose group, middle-dose group, high-dose group and western medicine group decreased significantly (P0.05). Compared with the model group, the activity of SOD in the low, medium and high dose group and the western medicine group was increased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the activity of SOD in the middle, high-dose group and western medicine group was increased, and the difference was statistically significant (P0.05). Compared with the middle dose group, the activity of SOD in the high dose and the western medicine group was increased, and the difference was statistically significant (P0.05). Compared with the western medicine group, the activity of SOD in high-dose group was increased, and the difference was statistically significant (P0.05). (3) Compared with the sham-operation group, the content of MDA in the remaining groups increased and the difference was statistically significant (P0.05). Compared with the model group, the content of MDA in the low dose group was decreased, but the difference was not significant (P0.05). The content of MDA in the middle and high dose group and the western medicine group was significantly decreased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the content of MDA in the middle and high-dose group and the western medicine group was significantly decreased, and the difference was statistically significant (P0.05). Compared with the middle-dose group, the content of MDA in the high-dose group and the western medicine group was decreased, and the difference was statistically significant (P0.05). Compared with the western medicine group, the content of MDA in the high-dose group was decreased, but the difference was not significant (P0.05). (4) The content of BDNF in the other groups increased with the sham-operation group, and the difference was significant (P0.05). Compared with the model group, the expression of BDNF in the low-dose group increased, but the difference was not significant (P0.05); the expression of BDNF in the middle and high-dose group and the western medicine group was significantly increased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the expression of BDNF in the middle-dose group increased, but the difference was not significant (P0.05); the expression of BDNF in the high-dose group and the western medicine group was significantly increased, and the difference was statistically significant (P0.05). The expression of BDNF in the high-dose group and the western medicine group was significantly increased compared with the middle-dose group (P0.05). There was no significant difference in BDNF in the high-dose group and the western medicine group (P0.05). (5) The content of NSE in the model group was significantly higher than that of the sham operation group (P0.05). The levels of NSE in the high-dose group and the western medicine group were not significant (P0.05). Compared with the model group, the content of NSE in the low-dose group was not significant, the difference was not significant (P0.05), and the levels of NSE in the middle and high dose groups and the western medicine group were lower, and the difference was statistically significant (P0.05). Compared with the low-dose group, the levels of NSE in the middle and high dose group and the western medicine group were lower, and the difference was statistically significant (P0.05). Compared with the middle-dose group, the levels of NSE in the high-dose group and the western medicine group were lower, and the difference was statistically significant (P0.05). There was no significant difference in the content of NSE in the high-dose group and the western medicine group (P0.05). (6) Compared with the sham operation group, the water content of the brain tissue of the model group and the rest group was increased, and the difference was statistically significant (P0.05). Compared with the model group, the water content of the brain tissue of the low-dose group, the middle-dose group, the high-dose group and the western medicine group were all decreased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the water content in the brain of the middle-dose group, the high-dose group and the western medicine group was decreased, and the difference was statistically significant (P0.05). Compared with the middle-dose group, the water content of the brain tissue of the high-dose group and the western medicine group was decreased, and the difference was statistically significant (P0.05). The changes of water content in the brain tissue of the high-dose group and the western medicine group were not significant (P0.05). (7) The results of light microscope showed that the cell structure of the sham-operated group was intact, the nucleus and the cytoplasm of the sham-operated group were intact, and the abnormal changes were not found. In the model group, edema, loose tissue, necrosis of the nerve cells and small amount of inflammatory cell infiltration around the hematoma were observed in the neurons and the cell gap. In the low, medium and high dose group, different degree of brain edema and inflammatory cell infiltration were found, but with the increase of the dose, the degree of edema and the inflammatory response were gradually decreased. The structure of the nerve cells around the hematoma in the western medicine group was relatively complete, and no obvious cerebral edema and inflammatory cell infiltration were found. Conclusion:1. Tongqiao Huoxue Decoction can improve the neurological deficit score of the cerebral hemorrhage rats, relieve the cerebral edema, improve the degeneration and necrosis of the nerve cells and the infiltration of the inflammatory cells around the hematoma after the cerebral hemorrhage. Possible mechanism: decrease the content of MDA, increase the SOD activity to remove oxygen free radicals, promote the expression of BDNF, reduce the NSE content, and thus protect the nerve cells. Tongqiao Huoxue Decoction has a certain amount of effect in preventing and treating cerebral hemorrhage in rats.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R277.7
本文编号：2488500
[Abstract]:Objective: To study the effect of Tongqiao Huoxue Decoction on the neurological function, NSE, BDNF, SOD, MDA and brain edema in rats with acute cerebral hemorrhage. Method:1. Group:84 male SD rats weighing (250 to 20) g were randomly divided into sham operation group, model group, Edarone group (short "Western medicine group"), Tongqiao Huoxue decoction group (short "high-dose group"), Tongqiao Huoxue decoction (short "mid-dose group"), Each group of the low-dose group (low-dose group ") of Tongqiao Huoxue Decoction (the low-dose group") was 14;2. Methods: The model was made by using the self-sampling method of the mouse tail, and the other operation steps of the sham-operated group were similar to that of the other groups. The method comprises the following steps of: administration of edaravone 1.5 mg/ kg/ day in the western medicine group into the abdominal cavity for intraperitoneal injection, wherein the high, middle and low dose groups are respectively provided with the Tongqiao Huoxue decoction 1.2 g/ Kg/ day, 0.6 g/ Kg/ day, 0.3 g/ Kg/ day, the sham operation group and the model group are given the same dose of normal saline for oral administration, each group is continuously administered for 14 days to form a model; and after the rats are finished, Each day, the neurological deficit score was scored, the Zea Longa 5 method was used to make the score, and the neurological deficit score was selected as the administration target; after the rats were awake, the sham-operation group and the model group continued to give the normal saline to the gastric administration, the low, medium, The high-dose group and the western medicine group were treated with the pre-model administration method for 1 week;4. Materials: The rats in each group after the last intragastric administration and after injection were treated with intraperitoneal anesthesia, the abdominal aorta was separated and the blood was punctured, and the heart was perfused immediately after blood collection and the brain tissue was decapitated. The results showed that the method of Zea Longa 5 was used to test the loss of nerve function, and the activity of SOD was detected by using the method of the method of using the method of the method of the method of the method of using the thiobarbituric acid, the content of the MDA was detected by the ELISA, the content of the BDNF was detected by the method of immunohistochemistry. The morphological changes of the nerve cells around the hematoma after cerebral hemorrhage were observed by light microscope. The water content of brain tissue was detected by dry and wet method. Results: (1) The score of the neurological deficit score was compared with that of the sham operation group. The scores of the neurological deficit in the other groups were increased and the difference was statistically significant (P0.05). Compared with the model group, the score of neurological deficit in the low-dose group was lower, but the difference was not significant (P0.05). In the middle and high dose group, the neurological deficit score of the western medicine group was significantly decreased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the functional deficit score of the middle and high dose group and the western medicine group was lower, and the difference was statistically significant (P0.05). Compared with the middle-dose group, the functional deficit score of the high-dose group and the western medicine group was lower, and the difference was statistically significant (P0.05). There was no significant difference in the score of neurological deficit in the high-dose group and the western medicine group (P0.05). (2) Compared with sham operation group, the activity of SOD in the group, low-dose group, middle-dose group, high-dose group and western medicine group decreased significantly (P0.05). Compared with the model group, the activity of SOD in the low, medium and high dose group and the western medicine group was increased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the activity of SOD in the middle, high-dose group and western medicine group was increased, and the difference was statistically significant (P0.05). Compared with the middle dose group, the activity of SOD in the high dose and the western medicine group was increased, and the difference was statistically significant (P0.05). Compared with the western medicine group, the activity of SOD in high-dose group was increased, and the difference was statistically significant (P0.05). (3) Compared with the sham-operation group, the content of MDA in the remaining groups increased and the difference was statistically significant (P0.05). Compared with the model group, the content of MDA in the low dose group was decreased, but the difference was not significant (P0.05). The content of MDA in the middle and high dose group and the western medicine group was significantly decreased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the content of MDA in the middle and high-dose group and the western medicine group was significantly decreased, and the difference was statistically significant (P0.05). Compared with the middle-dose group, the content of MDA in the high-dose group and the western medicine group was decreased, and the difference was statistically significant (P0.05). Compared with the western medicine group, the content of MDA in the high-dose group was decreased, but the difference was not significant (P0.05). (4) The content of BDNF in the other groups increased with the sham-operation group, and the difference was significant (P0.05). Compared with the model group, the expression of BDNF in the low-dose group increased, but the difference was not significant (P0.05); the expression of BDNF in the middle and high-dose group and the western medicine group was significantly increased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the expression of BDNF in the middle-dose group increased, but the difference was not significant (P0.05); the expression of BDNF in the high-dose group and the western medicine group was significantly increased, and the difference was statistically significant (P0.05). The expression of BDNF in the high-dose group and the western medicine group was significantly increased compared with the middle-dose group (P0.05). There was no significant difference in BDNF in the high-dose group and the western medicine group (P0.05). (5) The content of NSE in the model group was significantly higher than that of the sham operation group (P0.05). The levels of NSE in the high-dose group and the western medicine group were not significant (P0.05). Compared with the model group, the content of NSE in the low-dose group was not significant, the difference was not significant (P0.05), and the levels of NSE in the middle and high dose groups and the western medicine group were lower, and the difference was statistically significant (P0.05). Compared with the low-dose group, the levels of NSE in the middle and high dose group and the western medicine group were lower, and the difference was statistically significant (P0.05). Compared with the middle-dose group, the levels of NSE in the high-dose group and the western medicine group were lower, and the difference was statistically significant (P0.05). There was no significant difference in the content of NSE in the high-dose group and the western medicine group (P0.05). (6) Compared with the sham operation group, the water content of the brain tissue of the model group and the rest group was increased, and the difference was statistically significant (P0.05). Compared with the model group, the water content of the brain tissue of the low-dose group, the middle-dose group, the high-dose group and the western medicine group were all decreased, and the difference was statistically significant (P0.05). Compared with the low-dose group, the water content in the brain of the middle-dose group, the high-dose group and the western medicine group was decreased, and the difference was statistically significant (P0.05). Compared with the middle-dose group, the water content of the brain tissue of the high-dose group and the western medicine group was decreased, and the difference was statistically significant (P0.05). The changes of water content in the brain tissue of the high-dose group and the western medicine group were not significant (P0.05). (7) The results of light microscope showed that the cell structure of the sham-operated group was intact, the nucleus and the cytoplasm of the sham-operated group were intact, and the abnormal changes were not found. In the model group, edema, loose tissue, necrosis of the nerve cells and small amount of inflammatory cell infiltration around the hematoma were observed in the neurons and the cell gap. In the low, medium and high dose group, different degree of brain edema and inflammatory cell infiltration were found, but with the increase of the dose, the degree of edema and the inflammatory response were gradually decreased. The structure of the nerve cells around the hematoma in the western medicine group was relatively complete, and no obvious cerebral edema and inflammatory cell infiltration were found. Conclusion:1. Tongqiao Huoxue Decoction can improve the neurological deficit score of the cerebral hemorrhage rats, relieve the cerebral edema, improve the degeneration and necrosis of the nerve cells and the infiltration of the inflammatory cells around the hematoma after the cerebral hemorrhage. Possible mechanism: decrease the content of MDA, increase the SOD activity to remove oxygen free radicals, promote the expression of BDNF, reduce the NSE content, and thus protect the nerve cells. Tongqiao Huoxue Decoction has a certain amount of effect in preventing and treating cerebral hemorrhage in rats.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R277.7
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