維生素E干預(yù)對(duì)錳中毒小鼠學(xué)習(xí)記憶及神經(jīng)干細(xì)胞的影響
發(fā)布時(shí)間：2019-05-27 09:23
【摘要】：目的:采用神經(jīng)行為學(xué) 組織化學(xué)等方法研究錳中毒對(duì)小鼠海馬區(qū)神經(jīng)細(xì)胞的影響生素E對(duì)其干預(yù)作用,并探討其發(fā)生機(jī)制,為錳中毒的機(jī)理 預(yù)治療提供實(shí)驗(yàn)依據(jù)。方法:1、動(dòng)物 組:選用清潔級(jí)成年雄性健康的昆明小鼠75只,體重在18-22g(廣西醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物中心提供)。將選用的75只昆明小鼠按隨機(jī)對(duì)照方法分為5組,分別是對(duì)照組(CG)、生素E組(VG)、錳中毒組(MG)、錳加生素E組(MVG)、錳加普羅布考組(MPG),每組15只。2、制作錳中毒小鼠模型:分別對(duì)錳中毒、錳加生素E組、錳加普羅布考組3組小鼠腹腔注射氯化錳(Mncl2),劑量為20mg/kg,其余2組小鼠腹腔注射等容積的生理鹽水。每只小鼠注射的量為0.01ml/g,每天下午注射1次,連續(xù)注射4周。3、藥物灌胃:分別對(duì)生素E組、錳加生素E組行生素E藥物灌胃干預(yù),劑量為100mg/kg,對(duì)照組和錳中毒組行等容積的橄欖油灌胃,錳加普羅布考組予以普羅布考藥物灌胃,劑量為500mg/kg,每只小鼠灌胃的量是0.01ml/g,各組小鼠每天下午灌胃1次,連續(xù)給藥4周。各組小鼠在相同環(huán)境下飼養(yǎng),自由飲食。4、神經(jīng)行為學(xué)檢測(cè):4周后用Morris水迷宮檢測(cè)5組小鼠的逃避潛伏期和穿越平臺(tái)次數(shù),從而測(cè)定各組小鼠的學(xué)習(xí)記憶能力。5、免疫組織化學(xué)染色:用免疫組化的方法分別檢測(cè)每組小鼠海馬CA1區(qū)和DG區(qū)caspase-3、細(xì)胞色素C的表達(dá)和海馬DG區(qū)nestin的表達(dá)。計(jì)算所得免疫組織化學(xué)圖像的陽(yáng)性細(xì)胞數(shù),用統(tǒng)計(jì)軟件spss16.0和Graph Pad Prism5統(tǒng)計(jì)分析各組小鼠Morris水迷宮所得數(shù)據(jù)和免疫組化結(jié)果。結(jié)果:1、Morris水迷宮檢測(cè)結(jié)果:水迷宮檢測(cè)實(shí)驗(yàn)有定位航行實(shí)驗(yàn)和空間探索實(shí)驗(yàn)兩部分組成,定位航行實(shí)驗(yàn)中,與CG相比,MG小鼠的逃避潛伏期明顯延長(zhǎng),差異有統(tǒng)計(jì)學(xué)意義(P0.05);與MG相比,MVG和MPG小鼠的逃避潛伏期明顯縮短,差異有統(tǒng)計(jì)學(xué)意義(P0.05);MVG和MPG小鼠的逃避潛伏期相比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);CG和VG小鼠的逃避潛伏期相比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。在空間探索實(shí)驗(yàn)中,與CG相比,MG小鼠的穿越平臺(tái)的次數(shù)明顯減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05);而與MG比較,MVG和MPG小鼠穿越平臺(tái)次數(shù)明顯增多,差異有統(tǒng)計(jì)學(xué)意義(P0.05);MVG和MPG小鼠穿越平臺(tái)次數(shù)相比無(wú)明顯差別,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);CG和VG小鼠的穿越平臺(tái)次數(shù)無(wú)明顯差別,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。2、免疫組織化學(xué)細(xì)胞色素C的表達(dá):與CG相比,MG海馬CA1區(qū)和D G區(qū)細(xì)胞色素C的表達(dá)明顯增多,差異有統(tǒng)計(jì)學(xué)意義(P0.05);與MG相比,MVG和MPG小鼠細(xì)胞色素C的表達(dá)明顯減少,有統(tǒng)計(jì)學(xué)差異(P0.05);CG和VG之間海馬CA1區(qū)和DG區(qū)細(xì)胞色素C表達(dá)的比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);同時(shí)MVG和MPG間比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。3、免疫組織化學(xué)caspase-3的表達(dá):與CG相比較,MG海馬CA1區(qū)和DG區(qū)caspase-3陽(yáng)性細(xì)胞數(shù)明顯增多,差異有統(tǒng)計(jì)學(xué)意義(P0.05);與MG相比,MVG和MPG小鼠caspase-3的表達(dá)減少,差異有顯著性(P0.05);而CG和VG之間比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);同時(shí)MVG和MPG間比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。4、免疫組織化學(xué)nestin的表達(dá):與CG相比較,MG海馬齒狀回nestin的陽(yáng)性細(xì)胞數(shù)明顯減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05);NVG與MG相比,其nestin的表達(dá)明顯增多;而MG和MPG間nestin的表達(dá)無(wú)明顯差別(P0.05);同時(shí)CG和VG間比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:1、生素E和和普羅布考干預(yù)均可改善錳中毒小鼠的學(xué)習(xí)記憶能力;2.錳中毒可促進(jìn)小鼠腦內(nèi)細(xì)胞色素C的增多,上調(diào)caspase-3的表達(dá),從而降低小鼠的學(xué)習(xí)記憶能力;3、生素E和普羅布考可減少錳中毒小鼠細(xì)胞色素C和caspase-3的表達(dá),減少神經(jīng)細(xì)胞的凋亡;4、錳中毒可減少腦內(nèi)DG區(qū)神經(jīng)干細(xì)胞,生素E可增加錳中毒小鼠腦內(nèi)神經(jīng)干細(xì)胞的數(shù)量,而普羅布考對(duì)錳中毒小鼠腦內(nèi)神經(jīng)干細(xì)胞沒(méi)有影響;5、生素E可能主要通過(guò)抗氧化作用,來(lái)對(duì)改善錳中毒小鼠的學(xué)習(xí)記憶能力,同時(shí)生素E通過(guò)增加錳中毒腦內(nèi)DG區(qū)神經(jīng)干細(xì)胞數(shù)量,對(duì)錳中毒腦損傷的修復(fù)有一定的保護(hù)作用。
[Abstract]:Objective: To study the effect of manganese poisoning on the nerve cells in the hippocampus of mice by using the method of neurobehavioral histochemistry and the like, and to explore the mechanism of its occurrence and provide the experimental basis for the mechanism of manganese poisoning. Methods:1, animal group:75 male healthy Kunming mice were selected and the body weight was 18-22 g (provided by the experimental animal center of Guangxi Medical University). The selected 75 Kunming mice were divided into 5 groups according to the randomized controlled method. The control group (CG), the prebiotic E group (VG), the manganese poisoning group (MG), the manganese prebiotic E group (MVG) and the manganese plus probucol group (MPG) were divided into 5 groups. Manganese poisoning, manganese prebiotic E group, manganese plus probucol group 3 mice were injected intraperitoneally with manganese chloride (MnCl2) at a dose of 20 mg/ kg, and the rest of 2 mice were injected with saline in the same volume as the abdominal cavity. the injection amount of each mouse is 0.01ml/ g, and is injected every afternoon for 1 time, and is continuously injected for 4 weeks.3, the medicine is fed into the stomach by means of the intragastric administration of the prebiotic E, the dosage of which is 100 mg/ kg, the control group and the manganese poisoning group line, and the like, The dose of probucol was 500 mg/ kg and the amount of each mouse was 0.01 ml/ g. The mice were fed in the same environment and the free diet.4. Neurobehavioral test:4 weeks after 4 weeks, Morris water maze was used to detect the escape latency of 5 mice and the number of crossing platforms, so as to determine the learning and memory abilities of each group of mice. The expression of caspase-3, the expression of cytochrome C and the expression of nestin in the hippocampus of the hippocampus of each group were detected by immunohistochemistry. The numbers of the positive cells of the obtained immunohistochemical images were calculated, and the data and the immunohistochemical results of the Morris water maze of each group were statistically analyzed by using the statistical software sps16.0 and the Graph Pad Prim5. Results:1. The result of the Morris water maze test: The water maze test was made up of two parts: the positioning navigation experiment and the space exploration experiment. Compared with the CG, the escape latency of the MG mice was significantly prolonged compared with that of the CG (P0.05). The escape latency of MVG and MPG mice was significantly shortened, and the difference was significant (P0.05). The difference of the escape latency of MVG and MPG mice was not significant (P0.05). In the experiment of space exploration, the number of crossing platforms in MG mice was significantly decreased compared with CG (P0.05). Compared with MG, the number of crossing platforms of MVG and MPG mice increased significantly (P0.05). There was no significant difference in the number of crossing platforms of MVG and MPG mice (P0.05). There was no significant difference in the number of crossing platforms in CG and VG mice (P0.05). The expression of cytochrome C in the CA1 region and the D G region of MG was significantly increased, and the difference was significant (P0.05). Compared with MG, the expression of cytochrome C in the MVG and MPG mice was significantly reduced, and there was a statistical difference (P0.05). There was no significant difference in the expression of cytochrome C in the CA1 and DG regions of the hippocampus between CG and VG (P0.05), and the difference between MVG and MPG was not significant (P0.05). There was a significant difference in the expression of caspase-3 in MVG and MPG mice (P0.05). Compared with CG, the number of positive cells in the dentate gyrus of MG was significantly decreased, and the difference was significant (P0.05); the expression of nestin in MG and MPG was significantly higher than that of MG (P0.05); and the difference between CG and VG was not statistically significant (P0.05). Conclusion:1. Prebiotic E and propafenicol can improve the learning and memory ability of manganese-poisoned mice. Manganese poisoning can promote the increase of cytochrome C in the mouse brain, increase the expression of caspase-3, and reduce the learning and memory capacity of the mouse; and 3, the prebiotic E and the probucol can reduce the expression of the cytochrome C and the caspase-3 in the manganese-poisoned mice, and reduce the apoptosis of the nerve cells; and 4, Manganese poisoning can reduce the number of neural stem cells in the brain of the brain, which can increase the number of neural stem cells in the brain of the mice, and the probucol has no effect on the neural stem cells in the brain of the mice. So as to improve the learning and memory capacity of the manganese-poisoned mice, and meanwhile, the prebiotics E has a certain protective effect on the repair of the brain damage of the manganese poisoning by increasing the number of the neural stem cells in the DG region in the brain of the manganese poisoning.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R595
本文編號(hào)：2486035
[Abstract]:Objective: To study the effect of manganese poisoning on the nerve cells in the hippocampus of mice by using the method of neurobehavioral histochemistry and the like, and to explore the mechanism of its occurrence and provide the experimental basis for the mechanism of manganese poisoning. Methods:1, animal group:75 male healthy Kunming mice were selected and the body weight was 18-22 g (provided by the experimental animal center of Guangxi Medical University). The selected 75 Kunming mice were divided into 5 groups according to the randomized controlled method. The control group (CG), the prebiotic E group (VG), the manganese poisoning group (MG), the manganese prebiotic E group (MVG) and the manganese plus probucol group (MPG) were divided into 5 groups. Manganese poisoning, manganese prebiotic E group, manganese plus probucol group 3 mice were injected intraperitoneally with manganese chloride (MnCl2) at a dose of 20 mg/ kg, and the rest of 2 mice were injected with saline in the same volume as the abdominal cavity. the injection amount of each mouse is 0.01ml/ g, and is injected every afternoon for 1 time, and is continuously injected for 4 weeks.3, the medicine is fed into the stomach by means of the intragastric administration of the prebiotic E, the dosage of which is 100 mg/ kg, the control group and the manganese poisoning group line, and the like, The dose of probucol was 500 mg/ kg and the amount of each mouse was 0.01 ml/ g. The mice were fed in the same environment and the free diet.4. Neurobehavioral test:4 weeks after 4 weeks, Morris water maze was used to detect the escape latency of 5 mice and the number of crossing platforms, so as to determine the learning and memory abilities of each group of mice. The expression of caspase-3, the expression of cytochrome C and the expression of nestin in the hippocampus of the hippocampus of each group were detected by immunohistochemistry. The numbers of the positive cells of the obtained immunohistochemical images were calculated, and the data and the immunohistochemical results of the Morris water maze of each group were statistically analyzed by using the statistical software sps16.0 and the Graph Pad Prim5. Results:1. The result of the Morris water maze test: The water maze test was made up of two parts: the positioning navigation experiment and the space exploration experiment. Compared with the CG, the escape latency of the MG mice was significantly prolonged compared with that of the CG (P0.05). The escape latency of MVG and MPG mice was significantly shortened, and the difference was significant (P0.05). The difference of the escape latency of MVG and MPG mice was not significant (P0.05). In the experiment of space exploration, the number of crossing platforms in MG mice was significantly decreased compared with CG (P0.05). Compared with MG, the number of crossing platforms of MVG and MPG mice increased significantly (P0.05). There was no significant difference in the number of crossing platforms of MVG and MPG mice (P0.05). There was no significant difference in the number of crossing platforms in CG and VG mice (P0.05). The expression of cytochrome C in the CA1 region and the D G region of MG was significantly increased, and the difference was significant (P0.05). Compared with MG, the expression of cytochrome C in the MVG and MPG mice was significantly reduced, and there was a statistical difference (P0.05). There was no significant difference in the expression of cytochrome C in the CA1 and DG regions of the hippocampus between CG and VG (P0.05), and the difference between MVG and MPG was not significant (P0.05). There was a significant difference in the expression of caspase-3 in MVG and MPG mice (P0.05). Compared with CG, the number of positive cells in the dentate gyrus of MG was significantly decreased, and the difference was significant (P0.05); the expression of nestin in MG and MPG was significantly higher than that of MG (P0.05); and the difference between CG and VG was not statistically significant (P0.05). Conclusion:1. Prebiotic E and propafenicol can improve the learning and memory ability of manganese-poisoned mice. Manganese poisoning can promote the increase of cytochrome C in the mouse brain, increase the expression of caspase-3, and reduce the learning and memory capacity of the mouse; and 3, the prebiotic E and the probucol can reduce the expression of the cytochrome C and the caspase-3 in the manganese-poisoned mice, and reduce the apoptosis of the nerve cells; and 4, Manganese poisoning can reduce the number of neural stem cells in the brain of the brain, which can increase the number of neural stem cells in the brain of the mice, and the probucol has no effect on the neural stem cells in the brain of the mice. So as to improve the learning and memory capacity of the manganese-poisoned mice, and meanwhile, the prebiotics E has a certain protective effect on the repair of the brain damage of the manganese poisoning by increasing the number of the neural stem cells in the DG region in the brain of the manganese poisoning.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R595
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