【摘要】：目的:通過(guò)觀(guān)察通竅活血湯對(duì)急性腦出血大鼠神經(jīng)功能、NSE、BDNF、SOD、MDA、腦水腫的影響,初步探討通竅活血湯對(duì)大鼠腦出血防治的可能機(jī)制。方法:1.分組:取體重在(250±20)g的雄性SD大鼠84只,隨機(jī)分為假手術(shù)組、模型組、依達(dá)拉奉組(簡(jiǎn)稱(chēng)“西藥組”)、通竅活血湯高劑量組(簡(jiǎn)稱(chēng)“高劑量組”)、通竅活血湯中劑量組(簡(jiǎn)稱(chēng)“中劑量組”)、通竅活血湯低劑量組(簡(jiǎn)稱(chēng)“低劑量組”)每組各14只;2.造模方法:選用鼠尾自體采血注射法造模,假手術(shù)組除不注血外其余操作步驟同其他各組;3.給藥方法:西藥組給予依達(dá)拉奉1.5mg/kg/d腹腔注射,高、中、低劑量組分別給予通竅活血湯1.2g/Kg/d、0.6g/Kg/d、0.3g/Kg/d灌胃,假手術(shù)組、模型組給予相同劑量生理鹽水灌胃,各組連續(xù)給藥14天后造模;各組大鼠完成造模后,每天一次神經(jīng)功能缺損評(píng)分,選用Zea Longa5分制法進(jìn)行,選取神經(jīng)功能缺失評(píng)分在1-3分者作為給藥對(duì)象;大鼠清醒后,假手術(shù)組、模型組繼續(xù)給予生理鹽水灌胃,低、中、高劑量組和西藥組同造模前給藥方法繼續(xù)給藥1周;4.取材:末次灌胃及注射給藥后各組大鼠均采用腹腔麻醉法,分離腹主動(dòng)脈并穿刺采血;采血后立即心臟灌注并斷頭取腦組織。5.指標(biāo)檢測(cè):Zea Longa5分制法進(jìn)行神經(jīng)功能缺失評(píng)分,SOD活性的檢測(cè)按照說(shuō)明采用黃嘌呤氧化酶法,MDA含量的檢測(cè)采用硫代巴比妥酸法;ELISA法檢測(cè)NSE含量;免疫組化法檢測(cè)BDNF含量,光鏡觀(guān)察腦出血后血腫周?chē)窠?jīng)細(xì)胞形態(tài)變化。采用干濕重法檢測(cè)腦組織含水量。結(jié)果:(1)神經(jīng)功能缺失評(píng)分比較:與假手術(shù)組比較,其余各組神經(jīng)功能缺失評(píng)分均升高,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。與模型組比較,低劑量組神經(jīng)功能缺失評(píng)分有所降低,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),中、高劑量組、西藥組神經(jīng)功能缺失評(píng)分明顯降低,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。與低劑量組比較,中、高劑量組、西藥組神經(jīng)功能缺失評(píng)分降低,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。與中劑量組比較,高劑量組、西藥組神經(jīng)功能缺失評(píng)分降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。高劑量組與西藥組比較,神經(jīng)功能缺失評(píng)分變化不明顯,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。(2)SOD活性比較:與假手術(shù)組比較,模型組、低劑量組、中劑量組、高劑量組、西藥組SOD活性下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與模型組比較,低、中、高劑量組及西藥組SOD活性有所增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與低劑量組比較,中、高劑量組及西藥組的SOD活性增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與中劑量組比較,高劑量和西藥組SOD活性增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與西藥組比較,高劑量組SOD活性增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。(3)MDA含量比較:與假手術(shù)組比較,其余各組MDA含量均增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與模型組比較,低劑量組MDA含量有所減少,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),中、高劑量組及西藥組MDA含量明顯減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與低劑量組比較,中、高劑量組及西藥組MDA含量明顯減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與中劑量組比較,高劑量組和西藥組MDA含量減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與西藥組比較,高劑量組MDA含量有所減少,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。(4)BDNF含量比較:與假手術(shù)組比較,其他各組的BDNF含量均增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與模型組比較,低劑量組BDNF表達(dá)有所增加,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);中、高劑量組及西藥組BDNF表達(dá)明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與低劑量組比較,中劑量組BDNF表達(dá)有所增加,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);高劑量組及西藥組BDNF表達(dá)明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與中劑量組比較,高劑量組及西藥組BDNF表達(dá)明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。高劑量組與西藥組比較,BDNF變化不明顯,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。(5)NSE含量比較:與假手術(shù)組比較,模型組、低、中劑量組NSE含量均顯著增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05);高劑量組及西藥組NSE含量增加不明顯,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。與模型組比較,低劑量組NSE含量變化不明顯,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),中、高劑量組及西藥組NSE含量降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與低劑量組比較,中、高劑量組及西藥組NSE含量降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與中劑量組比較,高劑量組、西藥組NSE含量降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。高劑量組與西藥組比較,NSE含量變化不明顯,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。(6)腦含水量比較:與假手術(shù)組比較,模型組及其余各給藥組腦組織含水量均增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與模型組比較,低劑量組、中劑量組、高劑量組及西藥組腦組織含水量均減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與低劑量組比較,中劑量組、高劑量組及西藥組腦組織含水量減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與中劑量組比較,高劑量組及西藥組腦組織含水量減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。高劑量組與西藥組比較,腦組織含水量變化不明顯,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。(7)光鏡結(jié)果:光鏡下觀(guān)察,假手術(shù)組神經(jīng)細(xì)胞結(jié)構(gòu)完整,胞核、胞漿染色均勻,未見(jiàn)異常改變。模型組可見(jiàn)神經(jīng)元及細(xì)胞間隙發(fā)生水腫,組織結(jié)構(gòu)疏松,神經(jīng)細(xì)胞壞死,血腫周?chē)猩倭康难装Y細(xì)胞浸潤(rùn)。低、中、高劑量組可見(jiàn)不同程度的腦組織水腫及炎癥細(xì)胞浸潤(rùn),但隨著給藥劑量的增高,水腫程度及炎癥反應(yīng)逐漸降低。西藥組血腫周?chē)窠?jīng)細(xì)胞結(jié)構(gòu)排列相對(duì)完整,未見(jiàn)明顯的腦水腫及炎癥細(xì)胞浸潤(rùn)。結(jié)論:1.通竅活血湯可改善腦出血大鼠神經(jīng)功能缺失評(píng)分,減輕腦水腫,改善腦出血后血腫周?chē)窠?jīng)細(xì)胞變性壞死及炎癥細(xì)胞浸潤(rùn)。2.可能的機(jī)制:降低MDA含量,提高SOD活性達(dá)到清除氧自由基;促進(jìn)BDNF的表達(dá),降低NSE含量,從而保護(hù)神經(jīng)細(xì)胞。3.通竅活血湯在防治大鼠腦出血作用方面具有一定量效關(guān)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.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R277.7

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