發(fā)布時(shí)間：2018-03-25 14:12

  本文選題：腦缺血　切入點(diǎn)：益氣活血法　出處：《安徽中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：1目的本課題以中醫(yī)學(xué)“腎精-腦髓”和“氣血-血瘀”相關(guān)理論為依據(jù),將Wnt/-catenin信號(hào)調(diào)控做為研究切入點(diǎn),采用MCAO/R動(dòng)物模型,主要通過益氣活血法代表方腦絡(luò)欣通方和補(bǔ)腎生髓法代表方補(bǔ)腎生髓方,對(duì)腦缺血再灌注大鼠進(jìn)行干預(yù),觀察模型大鼠缺血側(cè)海馬及額頂葉皮質(zhì)區(qū)GSK3α、Sfrp1和Wnt9a蛋白表達(dá)的影響,最后通過實(shí)驗(yàn)結(jié)果和相關(guān)實(shí)驗(yàn)數(shù)據(jù),從分子生物學(xué)角度對(duì)益氣活血法和補(bǔ)腎生髓法治療缺血性中風(fēng)的機(jī)制和原理進(jìn)行闡述和分析。2方法首先選擇健康Sprague-Dawley(SD)的雄性大鼠,總共56只,月齡4個(gè)月左右,體重在280-320g之間。隨機(jī)將此56只實(shí)驗(yàn)大鼠分為4組,分別記為假手術(shù)組、模型組、益氣活血法組和補(bǔ)腎生髓法組,每組14只,并為每組大鼠進(jìn)行標(biāo)記,將每組標(biāo)記的大鼠再隨機(jī)分為2組,記為3d、7d組,每組7只。采用改良線栓法建立局灶性右側(cè)腦缺血大鼠模型,同時(shí)將益氣活血法代表方腦絡(luò)欣通方以及補(bǔ)腎生髓法代表方補(bǔ)腎生髓方,取中等劑量:8.54g/kg、12.87g/kg,對(duì)大鼠進(jìn)行灌胃給藥。益氣活血法組和補(bǔ)腎生髓法組在復(fù)制MCAO/R模型前0.5h灌胃給藥1次,以后于每天上午9:00和下午4:00分別灌胃一次,持續(xù)給藥3d、7d,假手術(shù)組和模型組給予同等劑量的生理鹽水灌胃。選取大鼠新鮮缺血側(cè)海馬及額頂葉皮質(zhì),應(yīng)用Western-Blot技術(shù)檢測(cè)GSK3α、Sfrp1和Wnt9a蛋白表達(dá)并加以分析,掃描顯色后的底片,運(yùn)用專業(yè)軟件Image J對(duì)圖像進(jìn)行灰度分析,SPSS20.0處理所得數(shù)據(jù),所得結(jié)果采用VISIO 2007軟件進(jìn)行圖表繪制。3結(jié)果Western Blot結(jié)果顯示:GSK3α條帶位于46/51kda處,Sfrp1條帶位于33kda處,Wnt9a條帶位于37kda處,Actin條帶位于42/44kda處。益氣活血法3d組和補(bǔ)腎生髓法3d組對(duì)腦缺血再灌注大鼠缺血側(cè)海馬區(qū)GSK3α、Sfrp1和Wnt9a蛋白表達(dá)的影響:與假手術(shù)組比較,模型組三種蛋白的相對(duì)表達(dá)量均明顯升高(P0.01);與模型組比較,益氣活血法組和補(bǔ)腎生髓法組三種蛋白表達(dá)均明顯降低(P0.01);與補(bǔ)腎生髓法組比較,益氣活血法組三種蛋白表達(dá)均明顯降低(P0.01)。額頂葉皮質(zhì)區(qū)與海馬區(qū)結(jié)果一致。益氣活血法7d組和補(bǔ)腎生髓法7d組對(duì)腦缺血再灌注大鼠缺血側(cè)海馬區(qū)GSK3α、Sfrp1和Wnt9a蛋白表達(dá)的影響:與假手術(shù)組比較,模型組三種蛋白表達(dá)均明顯升高(P0.01);與模型組比較,益氣活血法組和補(bǔ)腎生髓法組三種蛋白表達(dá)均明顯降低(P0.01);兩種復(fù)方無顯著差異(P0.05)。額頂葉皮質(zhì)區(qū)與海馬區(qū)結(jié)果一致。4結(jié)論兩種中藥復(fù)方能通過抑制缺血側(cè)海馬及皮質(zhì)GSK3α、Sfrp1和Wnt9a蛋白表達(dá),調(diào)節(jié)Wnt/-catenin信號(hào)通路,促進(jìn)局灶性腦缺血再灌注損傷腦組織修復(fù)。
[Abstract]:Objective based on the theories of "kidney essence and brain marrow" and "qi, blood and blood stasis" in traditional Chinese medicine (TCM), the Wnt/-catenin signal regulation was taken as the starting point and the animal model of MCAO/R was used. The effects of GSK3 偽 -Sfrp1 and Wnt9a protein expression in hippocampus and frontal parietal cortex of model rats were observed by invigorating qi and activating blood circulation method on behalf of Nao Luo Xin Tong Fang and Bushen Shengmai Fang on behalf of Bushen Shengmai recipe, and Bushen Shengmai Fang on behalf of Nao Luo Xin Tong Fang and Bushen Sheng Mei Fang on behalf of supplementing Qi and activating Blood Circulation. Finally, through the experimental results and related experimental data, from the molecular biological point of view, the mechanism and principle of Yiqi Huoxue method and Bushen Shengmai method in treating ischemic apoplexy were expounded and analyzed. Firstly, the healthy Sprague-Dawley SD male rats were selected, with 56 rats in total. The 56 experimental rats were randomly divided into four groups: sham operation group, model group, invigorating qi and activating blood circulation group and tonifying kidney and generating marrow group, 14 rats in each group. Each group of labeled rats was randomly divided into 2 groups, which were divided into 3 days and 7 days group with 7 rats in each group. The model of focal right cerebral ischemia was established by modified thread occlusion method. At the same time, tonifying Qi and activating Blood Circulation method on behalf of Nao Luo Xin Tong Fang and Bushen Shengli method on behalf of Bushen Shengmai recipe were taken at a moderate dose of 8.54 g / kg or 12.87 g / kg. The rats in the Yiqi Huoxue method group and Bushen Shengmai method group were given orally once 0.5 hours before the MCAO/R model was established. The rats in sham operation group and model group were given the same dose of normal saline at 9:00 and 4:00 for 3 days and 7 days respectively. The fresh ischemic hippocampus and frontal parietal cortex of rats were selected. The expression and analysis of GSK3 偽 -Sfrp1 and Wnt9a protein were detected by Western-Blot technique. The negative films were scanned and processed by SPSS 20.0 with the professional software Image J. The results were drawn by VISIO 2007 software. 3 results Western Blot results showed that the Western Blot showed that the Sfrp1 band was located at the 46/51kda, the Wnt9a band was located at the 37kda, the actin band was located at the 42/44kda, the 3d group of supplementing qi and activating blood circulation method and the 3d group of the method of reinforcing kidney and generating marrow. Effects of GSK3 偽 -Sfrp1 and Wnt9a protein expression in the hippocampus of ischemic rats after cerebral ischemia-reperfusion: compared with sham-operated group, Compared with the model group, the relative expression of three kinds of proteins in the model group was significantly higher than that in the model group, and compared with the model group, the expression of the three proteins in the Yiqi Huoxue group and the tonifying kidney group was significantly lower than that in the Bushen group. The expression of three kinds of protein in Yiqi and Huoxue treatment group was significantly decreased, and the results of frontal parietal cortex and hippocampus were consistent. The expression of GSK3 偽 Sfrp1 and Wnt9a protein in the ischemic hippocampus of rats with cerebral ischemia and reperfusion was observed in 7 days group of supplementing qi and activating blood circulation method and 7 days group of tonifying kidney and generating marrow method. Effects: compared with the sham-operated group, The expression of three proteins in the model group was significantly higher than that in the model group. The expression of three kinds of protein was significantly decreased in the two groups, and there was no significant difference between the two groups. The results of frontal parietal cortex and hippocampus were consistent with that of hippocampus. Conclusion the two Chinese herbs can inhibit the ischemic hippocampus and the skin of the ischemic side. The expression of Sfrp1 and Wnt9a proteins in the cytoplasm of GSK3 偽 -Sfrp1, Regulate Wnt/-catenin signal pathway and promote brain tissue repair after focal cerebral ischemia reperfusion injury.
【學(xué)位授予單位】：安徽中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R255.2

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