本文選題：清腦益元湯 + 腦缺血損傷　； 參考：《廣西中醫(yī)藥大學》2016年碩士論文

【摘要】：目的:觀察清腦益元湯對大鼠腦缺血損傷缺血側(cè)皮層及海馬區(qū)VEGF、PDGF表達的影響,探討清腦益元湯對大鼠腦缺血損傷神經(jīng)元保護的作用機制。方法:將成年健康雄性SD大鼠192只,按隨機數(shù)字表分為兩大組,每組96只,繼續(xù)隨機分為4個亞組:空白組(A組6只)、假手術組(B組30只)、模型對照組(C組30只)、清腦益元組(D組30只)。假手術組、模型對照組及清腦益元組均按缺血損傷后1d、3d、7d、14d、28d五個處死時間點分為5個亞組(n=6),并分別作如下處理:空白正常飼養(yǎng),灌胃等體積蒸餾水;假手術組單純手術而未插線動物,術后正常飼養(yǎng),灌胃等體積蒸餾水;模型對照組灌胃等體積蒸餾水7d后,麻醉,行MACO術,術后制成大鼠急性腦缺血損傷大鼠模型,待大鼠麻醉清醒后,繼續(xù)予生理鹽水灌胃至各處死時間點。清腦益元組灌胃等體積清腦益元7d后操作同模型對照組,待大鼠麻醉清醒后,繼續(xù)予中藥灌胃至大鼠各處死時間點。參考Longa的5分制法對大鼠進行神經(jīng)功能缺損評分,并按各時間點進行標本采集。取材后,一組應用實時定量逆轉(zhuǎn)錄聚合酶鏈反應(Real Time PCR)檢測大鼠缺血側(cè)皮層及海馬區(qū)VEGF的表達,另一組應用蛋白質(zhì)印跡法(Western Blot)檢測大鼠缺血側(cè)皮層及海馬區(qū)PDGF的表達。結(jié)果:1.RT-PCR檢測結(jié)果:與假手術組比較,模型組腦組織VEGF的mRNA表達在各時間點均有所升高,但統(tǒng)計學比較無顯著差異。與模型組比較,清腦益元組VEGF mRNA表達在各時間點均顯著增高。清腦益元組早期(第1天)可見VEGF表達增高,在早期(第3天)就明顯升高,并逐漸升高第14天達到高峰;在模型對照組VEGF表達成緩慢增長的趨勢,差異有統(tǒng)計學意義(P0.05)。2.蛋白質(zhì)印跡法結(jié)果:與模型組比較,清腦益元組PDGF的表達在各時間點均顯著增高。清腦益元組早期(第1天)可見PDGF的明顯增高,在早期(第3天)呈稍放緩但仍很明顯的增高趨勢,并逐漸升高第14天達到高峰;在模型組PDGF表達成緩慢增長的趨勢,差異有統(tǒng)計學意義(P0.05)。結(jié)論:清腦益元湯對大鼠腦缺血損傷神經(jīng)元的保護作用可能與VEGF、PDGF表達的增加有關。
[Abstract]:Objective: to observe the effect of Qingnao Yiyuan decoction (QNYD) on the expression of VEGF PDGF in ischemic cortex and hippocampus of rats with cerebral ischemia injury, and to explore the protective mechanism of Qingnao Yiyuan decoction (QIYD) on neurons injured by cerebral ischemia in rats. Methods: 192 adult male SD rats were randomly divided into two groups, 96 rats in each group. They were randomly divided into four subgroups: group A (n = 6), group B (n = 30), group C (n = 30) and group D (n = 30). Sham-operation group, model control group and Qingnao Yiyuan group were divided into 5 subgroups according to five death time points: normal blank feeding and gastric distilled water. In the sham operation group, the rats were fed normally and fed with distilled water of the same volume after operation, and the rats in the model control group were anesthetized and treated with MACO for 7 days after the rats were fed with distilled water of the same volume. The model of acute cerebral ischemia injury in rats was made after operation. After the rats were anesthetized and awake, the rats were gavaged with normal saline until all the time points were put to death. The rats in the Qingnao Yiyuan group were treated with the same model control group after 7 days of oral administration of the same volume of Qingnao Yiyuan. After the rats were anesthetized and awake, the rats were continued to be gavaged with traditional Chinese medicine to the death time points of the rats. According to the 5-point method of Longa, the neurological impairment score of rats was evaluated, and the specimens were collected according to the time points. The expression of VEGF in ischemic cortex and hippocampus was detected by real time PCR and Western blot was used to detect the expression of PDGF in ischemic cortex and hippocampus of rats. Results 1. The results of RT-PCR: compared with sham operation group, the expression of VEGF mRNA in brain tissue of model group was increased at all time points, but there was no significant difference between the two groups. Compared with model group, VEGF mRNA expression in Qingnao Yiyuan group was significantly increased at all time points. In Qingnao Yiyuan group, the expression of VEGF increased at the early stage (on the first day), increased obviously at the early stage (on the third day), and gradually increased to the peak on the 14th day, while in the model control group, the expression of VEGF increased slowly, and the difference was statistically significant (P 0.05). Results of Western blotting: compared with model group, PDGF expression in Qingnao Yiyuan group was significantly higher than that in model group. In Qingnao Yiyuan group, PDGF increased significantly at the early stage (day 1), but decreased slightly at the early stage (day 3), and reached its peak on the 14th day, and the expression of PDGF increased slowly in the model group. The difference was statistically significant (P 0.05). Conclusion: the protective effect of Qingnao Yiyuan decoction on neurons injured by cerebral ischemia may be related to the increase of VEGF PDGF expression.
【學位授予單位】：廣西中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R277.7

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