三重腦震蕩鼠認(rèn)知行為障礙與腦多巴胺能神經(jīng)元變化研究
發(fā)布時(shí)間：2018-08-21 07:48
【摘要】： 目的：建立三重腦震蕩(multiple cerebral concussion，MCC)大鼠模型，觀察MCC大鼠腦多巴胺能神經(jīng)元、神經(jīng)纖維變化的規(guī)律，探討與MCC大鼠認(rèn)知行為障礙相關(guān)腦區(qū)的多巴胺能神經(jīng)元變化，以了解三重腦震蕩鼠認(rèn)知行為障礙的神經(jīng)生物學(xué)基礎(chǔ)。方法：應(yīng)用金屬單擺打擊裝置復(fù)制大鼠MCC模型48只，大鼠隨機(jī)分為1天、2天、4天、8天、16天和24天組共6個(gè)三重腦震蕩組(n=8)，另設(shè)一正常對(duì)照組(n=8)。Morris水迷宮(MWM)分別于傷后1～7天、8～14天、15～21天連續(xù)7天檢測(cè)大鼠的學(xué)習(xí)記憶能力。用抗TH和抗DBH單克隆抗體行免疫組化S-P法，聯(lián)合標(biāo)記中腦黑質(zhì)致密區(qū)、腹側(cè)被蓋區(qū)、前額葉皮質(zhì)、前扣帶皮質(zhì)、尾殼核、伏隔核、嗅結(jié)節(jié)、額頂葉皮質(zhì)、后扣帶皮質(zhì)和內(nèi)嗅皮質(zhì)多巴胺能神經(jīng)。用光鏡對(duì)大鼠中腦黑質(zhì)致密區(qū)、腹側(cè)被蓋區(qū)的TH陽(yáng)性細(xì)胞進(jìn)行觀察和計(jì)數(shù)，并對(duì)大腦前額葉皮質(zhì)、前扣帶皮質(zhì)、尾殼核、伏隔核、嗅結(jié)節(jié)、額頂葉皮質(zhì)、后扣帶皮質(zhì)和內(nèi)嗅皮質(zhì)的TH陽(yáng)性纖維密度和強(qiáng)弱進(jìn)行觀察和半定量檢測(cè)；用圖像分析系統(tǒng)分別對(duì)上述腦區(qū)的TH陽(yáng)性細(xì)胞和神經(jīng)纖維進(jìn)行定量分析。結(jié)果：(1)在Morris水迷宮測(cè)試中，，MCC后16d組的第9、10、13、14天與對(duì)照組相比差異具有顯著性(P＜0.05)：8d、24d組找到平臺(tái)的時(shí)間與對(duì)照組相比有所延長(zhǎng)，但差異無(wú)顯著性(P＞0.05)。在無(wú)平臺(tái)探測(cè)實(shí)驗(yàn)中，與對(duì)照組相比，MCC后8d、16d組大鼠在原有平臺(tái)第三象限停留時(shí)間明顯減少，差異具有顯著性(P＜0.05)；MCC后24d組在原有平臺(tái)第三象限停留時(shí)間稍有減少，差異無(wú)顯著性(P＞0.05)。(2)MCC后腦黑質(zhì)致密區(qū)和腹側(cè)被蓋區(qū)TH陽(yáng)性細(xì)胞計(jì)數(shù)分析結(jié)果：在黑質(zhì)致密區(qū)TH陽(yáng)性細(xì)胞數(shù)以傷后1天開始升高，8天達(dá)到高峰，與對(duì)照組相比4天、8天組有顯著性差異(p＜0.05)；與對(duì)照組相比，腹側(cè)被蓋區(qū)2天、4天、8天、16天組有顯著性差異(p＜0.05)，以4天組TH陽(yáng)性細(xì)胞數(shù)達(dá)到高峰，24天組基本恢復(fù)。(3)MCC后腦黑質(zhì)致密區(qū)和腹側(cè)被蓋區(qū)TH陽(yáng)性細(xì)胞圖像分析結(jié)果：TH免疫反應(yīng)陽(yáng)性表達(dá)在黑質(zhì)致密區(qū)以傷后1天開始升高，4天達(dá)到高峰，與對(duì)照組相比有顯著性差異(p＜0.01)，8天開始減弱，與對(duì)照組相比仍有顯著性差異(p＜0.05)，24天組基本恢復(fù)。與對(duì)照組相比，腹側(cè)被蓋區(qū)2天、4天、8天、16天組有顯著性差異(p＜0.05)，以2天組陽(yáng)性反應(yīng)最強(qiáng)(p＜0.01)，24天組基本恢復(fù)。(4)大鼠腦前額葉皮質(zhì)、前扣帶皮質(zhì)、尾殼核、伏隔核、嗅結(jié)節(jié)、額頂葉皮質(zhì)、后扣帶皮質(zhì)和內(nèi)嗅皮質(zhì)TH陽(yáng)性纖維密度和強(qiáng)弱半定量分析結(jié)果：與對(duì)照組相比，傷后1天、4天、16天組伏隔核TH陽(yáng)性纖維密度和強(qiáng)弱明顯增強(qiáng)(p＜0.008)。嗅結(jié)節(jié)于傷后1天、8天組，TH免疫反應(yīng)陽(yáng)性纖維的密度和強(qiáng)弱高于對(duì)照組，差異具有顯著性(p≤0.008)。內(nèi)嗅皮質(zhì)于傷后8天組，TH免疫反應(yīng)陽(yáng)性纖維的密度和強(qiáng)弱明顯高于對(duì)照組(p＜0.008)。前額葉皮質(zhì)、前扣帶皮質(zhì)、尾殼核、額頂葉皮質(zhì)、后扣帶皮質(zhì)TH免疫反應(yīng)陽(yáng)性纖維的密度和強(qiáng)弱有不同程度的增強(qiáng)，但與對(duì)照組相比無(wú)顯著差異(p＞0.008)。(5)MCC后不同腦區(qū)TH陽(yáng)性纖維圖像分析結(jié)果：傷后4天、8天組，前額葉皮質(zhì)多巴胺能神經(jīng)纖維免疫反應(yīng)陽(yáng)性顯著高于對(duì)照組(p＜0.05)，4天組反應(yīng)達(dá)高峰，24天組仍高于對(duì)照組。與對(duì)照組相比，各損傷組前扣帶皮質(zhì)顯著高于對(duì)照組(p≤0.01)，16天組反應(yīng)達(dá)高峰，24天組仍未恢復(fù)。傷后1天、4天、16天組，尾殼核TH陽(yáng)性纖維顯著高于對(duì)照組(p＜0.05)，1天組反應(yīng)達(dá)高峰，24天組仍高于對(duì)照組。與對(duì)照組相比，伏隔核TH陽(yáng)性纖維于傷后4天、8天、16天、24天組顯著增高(p＜0.01)，8天組反應(yīng)達(dá)高峰，24天組仍未恢復(fù)。嗅結(jié)節(jié)TH陽(yáng)性纖維1天、8天組與對(duì)照組相比有顯著差異(p＜0.05)，8天組反應(yīng)達(dá)高峰，24天組基本恢復(fù)。與對(duì)照組相比，各損傷組額頂葉皮質(zhì)TH陽(yáng)性纖維明顯增高(p＜0.05)，1天組反應(yīng)達(dá)高峰，24天組仍未恢復(fù)。后扣帶皮質(zhì)TH陽(yáng)性纖維于傷后1天、2天組與對(duì)照組相比有顯著差異(p＜0.05)，2天組反應(yīng)達(dá)高峰，24天組基本恢復(fù)。內(nèi)嗅皮質(zhì)于傷后1天、2天、8天組明顯高于對(duì)照組(p＜0.05)，2天組反應(yīng)達(dá)高峰，24天組仍未恢復(fù)。以上各部位免疫組化DBH反應(yīng)均為陰性。結(jié)論：1．金屬單擺式閉合性機(jī)械打擊裝置能成功建立三重腦震蕩大鼠模型；2．三重腦震蕩后大鼠出現(xiàn)中期學(xué)習(xí)記憶功能障礙；3．三重腦震蕩后，大鼠大腦皮質(zhì)、尾殼核、伏隔核、嗅結(jié)節(jié)、黑質(zhì)致密區(qū)、腹側(cè)被蓋區(qū)TH陽(yáng)性表達(dá)出現(xiàn)不同程度的增高。4．三重腦震蕩后大鼠行為學(xué)測(cè)試結(jié)果在不同時(shí)間點(diǎn)的變化與TH陽(yáng)性表達(dá)產(chǎn)物在不同時(shí)間點(diǎn)的變化趨勢(shì)基本一致。5．三重腦震蕩后大鼠認(rèn)知行為障礙可能與傷后腦內(nèi)多巴胺能神經(jīng)元變化有一定關(guān)系。
[Abstract]:AIM: To establish a rat model of multiple cerebral concussion (MCC), observe the changes of dopaminergic neurons and nerve fibers in the brain of MCC rats, and explore the changes of dopaminergic neurons in the brain regions associated with cognitive and behavioral disorders in MCC rats, so as to understand the neurobiological basis of cognitive and behavioral disorders in rats with MCC. METHODS: Forty-eight MCC rats were randomly divided into 6 triple concussion groups (n=8) and a normal control group (n=8). Morris water maze (MWM) was used to test the learning and memory abilities of rats 1-7 days, 8-14 days, 15-21 days after injury. Immunohistochemical S-P method was used to label the TH positive cells in substantia nigra compact area, ventral tegmental area, prefrontal cortex, anterior cingulate cortex, caudate putamen nucleus, nucleus accumbens, olfactory nodule, frontal parietal cortex, posterior cingulate cortex and endoolfactory cortex. The density and intensity of TH-positive fibers in prefrontal cortex, anterior cingulate cortex, caudate putamen nucleus, nucleus accumbens, olfactory tubercle, frontal-parietal cortex, posterior cingulate cortex and inner olfactory cortex were observed and counted, and the TH-positive cells and nerve fibers in these areas were quantitatively analyzed by image analysis system. Results: (1) In the Morris water maze test, there was significant difference between the control group and the MCC group on the 9th, 10th, 13th and 14th day after MCC (P The residence time in the third quadrant of the original platform was slightly decreased in the 24 days after MCC, but there was no significant difference (P > 0.05). (2) The number of TH positive cells in the substantia nigra compact area and the ventral tegmental area increased from 1 day after MCC. Compared with the control group, there were significant differences in the ventral tegmental area of 2 days, 4 days, 8 days and 16 days (p < 0.05), and the number of TH positive cells in the 4 days group reached the peak, and the number of TH positive cells in the 24 days group basically recovered. (3) The image scores of TH positive cells in the substantia nigra compact area and ventral tegmental area after MCC. Results: The expression of TH immunoreactivity in substantia nigra dense area increased from 1 day to 4 days after injury, and reached the peak at 4 days. There was significant difference between the control group and the control group (p < 0.01). There was a significant difference (p < 0.05). The TH positive fibers in the prefrontal cortex, anterior cingulate cortex, caudate putamen nucleus, nucleus accumbens, olfactory nodule, frontal parietal cortex, posterior cingulate cortex and inner olfactory cortex were the strongest in the 2-day group (p < 0.01) and recovered basically in the 24-day group. The density and intensity of TH-positive fibers in the nucleus accumbens were significantly increased (p < 0.008). The density and intensity of TH-immunoreactive fibers in the olfactory nodules were significantly higher than those in the control group on the 1st and 8th day after injury (p < 0.008). The density and intensity of TH-immunoreactive fibers in the inner olfactory cortex were significantly higher than those in the control group on the 8th day after injury (p < 0.008). The density and intensity of TH immunoreactive fibers in the cortex, anterior cingulate cortex, caudate putamen nucleus, frontal and parietal cortex, and posterior cingulate cortex were increased in varying degrees, but there was no significant difference between the two groups (p > 0.008). (5) The results of image analysis of TH immunoreactive fibers in different brain regions after MCC: the prefrontal cortex at 4 and 8 days after injury, the prefrontal cortex dopaminergic nerve fibers. Compared with the control group, the anterior cingulate cortex of each injury group was significantly higher than that of the control group (p < 0.01). The reaction peaked in the 16 days group and did not recover in the 24 days group. The TH positive fibers in the caudate putamen nucleus were significantly higher in the 1, 4 and 16 days group than that in the control group (p < 0.05). Compared with the control group, the TH-positive fibers in the nucleus accumbens increased significantly on the 4th, 8th, 16th and 24th day after injury (P < 0.01), the reaction peaked in the 8th day group and remained unchanged in the 24th day group. Compared with the control group, the TH-positive fibers in the frontal and parietal cortex of each injury group were significantly higher (p < 0.05), the reaction peaked in the first day group, but did not recover in the 24 day group. Immunohistochemical DBH reaction was negative in all the above parts. Conclusion: 1. Metal pendulum closed mechanical percussion device can successfully establish the rat model of triple cerebral concussion; 2. Middle-term learning after triple cerebral concussion in rats. Memory impairment; 3. After triple concussion, the TH positive expression in the cerebral cortex, caudate putamen, nucleus accumbens, olfactory nodule, substantia nigra compact area, ventral tegmental area increased in varying degrees. 4. Behavioral changes of rats after triple concussion at different time points and the trend of TH positive expression products at different time points. 5. Cognitive and behavioral disorders in rats after triple concussion may be related to the changes of dopaminergic neurons in the brain.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：D919
本文編號(hào)：2195036
[Abstract]:AIM: To establish a rat model of multiple cerebral concussion (MCC), observe the changes of dopaminergic neurons and nerve fibers in the brain of MCC rats, and explore the changes of dopaminergic neurons in the brain regions associated with cognitive and behavioral disorders in MCC rats, so as to understand the neurobiological basis of cognitive and behavioral disorders in rats with MCC. METHODS: Forty-eight MCC rats were randomly divided into 6 triple concussion groups (n=8) and a normal control group (n=8). Morris water maze (MWM) was used to test the learning and memory abilities of rats 1-7 days, 8-14 days, 15-21 days after injury. Immunohistochemical S-P method was used to label the TH positive cells in substantia nigra compact area, ventral tegmental area, prefrontal cortex, anterior cingulate cortex, caudate putamen nucleus, nucleus accumbens, olfactory nodule, frontal parietal cortex, posterior cingulate cortex and endoolfactory cortex. The density and intensity of TH-positive fibers in prefrontal cortex, anterior cingulate cortex, caudate putamen nucleus, nucleus accumbens, olfactory tubercle, frontal-parietal cortex, posterior cingulate cortex and inner olfactory cortex were observed and counted, and the TH-positive cells and nerve fibers in these areas were quantitatively analyzed by image analysis system. Results: (1) In the Morris water maze test, there was significant difference between the control group and the MCC group on the 9th, 10th, 13th and 14th day after MCC (P The residence time in the third quadrant of the original platform was slightly decreased in the 24 days after MCC, but there was no significant difference (P > 0.05). (2) The number of TH positive cells in the substantia nigra compact area and the ventral tegmental area increased from 1 day after MCC. Compared with the control group, there were significant differences in the ventral tegmental area of 2 days, 4 days, 8 days and 16 days (p < 0.05), and the number of TH positive cells in the 4 days group reached the peak, and the number of TH positive cells in the 24 days group basically recovered. (3) The image scores of TH positive cells in the substantia nigra compact area and ventral tegmental area after MCC. Results: The expression of TH immunoreactivity in substantia nigra dense area increased from 1 day to 4 days after injury, and reached the peak at 4 days. There was significant difference between the control group and the control group (p < 0.01). There was a significant difference (p < 0.05). The TH positive fibers in the prefrontal cortex, anterior cingulate cortex, caudate putamen nucleus, nucleus accumbens, olfactory nodule, frontal parietal cortex, posterior cingulate cortex and inner olfactory cortex were the strongest in the 2-day group (p < 0.01) and recovered basically in the 24-day group. The density and intensity of TH-positive fibers in the nucleus accumbens were significantly increased (p < 0.008). The density and intensity of TH-immunoreactive fibers in the olfactory nodules were significantly higher than those in the control group on the 1st and 8th day after injury (p < 0.008). The density and intensity of TH-immunoreactive fibers in the inner olfactory cortex were significantly higher than those in the control group on the 8th day after injury (p < 0.008). The density and intensity of TH immunoreactive fibers in the cortex, anterior cingulate cortex, caudate putamen nucleus, frontal and parietal cortex, and posterior cingulate cortex were increased in varying degrees, but there was no significant difference between the two groups (p > 0.008). (5) The results of image analysis of TH immunoreactive fibers in different brain regions after MCC: the prefrontal cortex at 4 and 8 days after injury, the prefrontal cortex dopaminergic nerve fibers. Compared with the control group, the anterior cingulate cortex of each injury group was significantly higher than that of the control group (p < 0.01). The reaction peaked in the 16 days group and did not recover in the 24 days group. The TH positive fibers in the caudate putamen nucleus were significantly higher in the 1, 4 and 16 days group than that in the control group (p < 0.05). Compared with the control group, the TH-positive fibers in the nucleus accumbens increased significantly on the 4th, 8th, 16th and 24th day after injury (P < 0.01), the reaction peaked in the 8th day group and remained unchanged in the 24th day group. Compared with the control group, the TH-positive fibers in the frontal and parietal cortex of each injury group were significantly higher (p < 0.05), the reaction peaked in the first day group, but did not recover in the 24 day group. Immunohistochemical DBH reaction was negative in all the above parts. Conclusion: 1. Metal pendulum closed mechanical percussion device can successfully establish the rat model of triple cerebral concussion; 2. Middle-term learning after triple cerebral concussion in rats. Memory impairment; 3. After triple concussion, the TH positive expression in the cerebral cortex, caudate putamen, nucleus accumbens, olfactory nodule, substantia nigra compact area, ventral tegmental area increased in varying degrees. 4. Behavioral changes of rats after triple concussion at different time points and the trend of TH positive expression products at different time points. 5. Cognitive and behavioral disorders in rats after triple concussion may be related to the changes of dopaminergic neurons in the brain.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：D919
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 柯尊記,姚志彬;空間學(xué)習(xí)記憶的行為模式及其分子神經(jīng)生物學(xué)基礎(chǔ)[J];解剖科學(xué)進(jìn)展;1998年01期
2 吳春云;于建云;林海英;郭澤云;許冰瑩;李坪;趙旭東;;腦震蕩對(duì)大鼠多巴胺能神經(jīng)元及神經(jīng)纖維的影響[J];解剖學(xué)雜志;2006年01期
本文編號(hào)：2195036
本文鏈接：http://sikaile.net/shekelunwen/gongan/2195036.html
最近更新
教材專著
