三重腦震蕩鼠多巴胺神經(jīng)與膽堿能神經(jīng)變化關(guān)系研究
發(fā)布時(shí)間：2018-08-18 12:13
【摘要】： 目的:建立三重腦震蕩(multiple cerebral concussion,MCC)大鼠模型,觀(guān)察MCC大鼠大腦多巴胺能神經(jīng)元和膽堿能神經(jīng)元、神經(jīng)纖維變化的規(guī)律。探討與MCC大鼠認(rèn)知行為相關(guān)腦區(qū)多巴胺能神經(jīng)元與膽堿能神經(jīng)元之間的相互關(guān)系,以了解三重腦震蕩鼠神經(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36只,大鼠隨機(jī)分為1天、2天、4天、8天、16天和24天組6個(gè)三重腦震蕩組(n=6),另設(shè)一正常對(duì)照組(n=6)。用抗TH和抗CHAT抗體進(jìn)行免疫組化雙標(biāo)S-P法檢測(cè)大鼠中腦黑質(zhì)致密區(qū)、前扣帶皮質(zhì)、尾殼核、伏隔核、前額葉皮質(zhì)、基底前腦、腦干網(wǎng)狀結(jié)構(gòu)、Meynert基底神經(jīng)核的多巴胺能神經(jīng)和膽堿能神經(jīng)表達(dá)變化及二者間的作用關(guān)系。用光鏡對(duì)大鼠中腦黑質(zhì)致密區(qū)、前扣帶皮質(zhì)、尾殼核、伏隔核、前額葉皮質(zhì)、基底前腦、腦干網(wǎng)狀結(jié)構(gòu)、Meynert基底神經(jīng)核的多巴胺能神經(jīng)和膽堿能神經(jīng)進(jìn)行記數(shù),并用圖像分析系統(tǒng)分別對(duì)上述腦區(qū)的TH和CHAT陽(yáng)性細(xì)胞、纖維進(jìn)行光密度分析。結(jié)果:(1)MCC后腦黑質(zhì)致密區(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(kāi)始上升,8天達(dá)高峰,與對(duì)照組相比2天組、4天組、8天組、16天組有顯著性差異(p＜0.05),24天組基本恢復(fù)。(2)MCC后腦黑質(zhì)致密區(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(kāi)始上升,8天達(dá)高峰,與對(duì)照組相比8天有顯著性差異(p＜0.05),24天組基本恢復(fù)。(3)大鼠腦前扣帶皮質(zhì)、尾殼核、伏隔核、前額葉皮質(zhì)和內(nèi)側(cè)隔核、斜角帶垂直支、水平支TH陽(yáng)性纖維光密度分析結(jié)果:與對(duì)照組相比,各損傷組TH陽(yáng)性纖維上升,8天組反應(yīng)達(dá)高峰,24天組基本恢復(fù)正常。尾殼核8天組,內(nèi)側(cè)隔核8天組、16天組,斜角帶垂直支8天組、16天組,斜角帶水平支4天組、16天組與對(duì)照組相比有統(tǒng)計(jì)學(xué)差異(p＜0.05)。(4)MCC后不同腦區(qū)CHAT陽(yáng)性細(xì)胞數(shù):腦干網(wǎng)狀結(jié)構(gòu)、內(nèi)側(cè)隔核(MS)、斜角帶垂直支(VDB)、左、右側(cè)斜角帶水平支(HDB)ChAT陽(yáng)性細(xì)胞數(shù)在MCC后1～24d明顯減少,然后逐漸增加至傷后24d組基本正常。與對(duì)照組比較,內(nèi)側(cè)隔核(MS)1～24d差異有明顯統(tǒng)計(jì)學(xué)意義(P＜0.05)。左、右斜角帶垂直支(VDB)在1d、2d、4d、8d有顯著差異,左、右側(cè)斜角帶水平支(HDB)在1d、2d、4d、有顯著差異(P＜0.05)。Meynert基底神經(jīng)核ChAT陽(yáng)性細(xì)胞數(shù)傷后1d、2d明顯減少,然后逐漸增加至傷后24d組基本正常。與對(duì)照組比較Meynert基底神經(jīng)核1d、2d差異有明顯統(tǒng)計(jì)學(xué)意義(P＜0.05)。左、右側(cè)相比差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)。(5)MCC后在基底節(jié)前腦ChAT陽(yáng)性細(xì)胞光密度值:ChAT陽(yáng)性細(xì)胞光密度值在MCC后1d～24d均呈現(xiàn)下降趨勢(shì),至4天稍有升高后,8天下降至最低。與對(duì)照組相比,內(nèi)側(cè)隔核(MS)于MCC后16d組具有統(tǒng)計(jì)學(xué)意義(P＜0.05)。左、右側(cè)斜角帶垂直支(VDB)、內(nèi)側(cè)隔核和右側(cè)斜角帶水平支(HDB)于MCC后8d組、16天和24天組相比較差異具有統(tǒng)計(jì)學(xué)意義(P＜O.05)。內(nèi)側(cè)隔核、左、右側(cè)斜角帶垂直支(VDB)至傷后24d組仍未恢復(fù)正常。左側(cè)斜角帶水平支(HDB)8d、右側(cè)斜角帶水平支(HDB)24d和對(duì)照組比較有統(tǒng)計(jì)學(xué)意義(P＜0.05)。左、右側(cè)Meynert基底神經(jīng)核在MCC后8d時(shí)稍有上升后在16d組出現(xiàn)降低,至24天組有所恢復(fù)正常。Meynert基底神經(jīng)核左右側(cè)相比差異均無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)。(6)MCC后腦干網(wǎng)狀結(jié)構(gòu)(BRF)ChAT陽(yáng)性細(xì)胞和TH陽(yáng)性纖維光密度值:腦干網(wǎng)狀結(jié)構(gòu)各組ChAT免疫反應(yīng)陽(yáng)性亦呈現(xiàn)下降趨勢(shì),至4天稍有上升后,8天下降至最低,至24d基本恢復(fù)正常。與對(duì)照組相比,在腦橋核平面1d到24d組均具有統(tǒng)計(jì)學(xué)意義(P＜0.05);而在斜方體核在傷后1d、2d、8d、16d組具有統(tǒng)計(jì)學(xué)意義(P＜0.05);橄欖核平面1d、2d、16d具有統(tǒng)計(jì)學(xué)意義(P＜0.05)。腦橋核、斜方體核和橄欖核平面在MCC后TH陽(yáng)性纖維1d～24d均呈現(xiàn)上升趨勢(shì),8天組達(dá)高峰。與對(duì)照組相比,腦橋核8d、斜方體核16d和橄欖核平面8d有統(tǒng)計(jì)學(xué)意義(p＜0.05)。結(jié)論:1.采用金屬單擺式機(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,大鼠大腦皮質(zhì)、尾殼核、伏隔核、黑質(zhì)致密區(qū)、內(nèi)側(cè)隔核、斜角帶、腦干核平面TH陽(yáng)性表達(dá)出現(xiàn)不同程度的上升,以傷后8天為重。3.MCC大鼠基底前腦、腦干網(wǎng)狀結(jié)構(gòu)膽堿能神經(jīng)元數(shù)目和活性均呈現(xiàn)下降趨勢(shì),以傷后8天組和16天組為重。4.三重腦震蕩后,大鼠大腦皮質(zhì)、基底前腦、腦干核平面多個(gè)與認(rèn)知記憶相關(guān)的腦區(qū)出現(xiàn)TH陽(yáng)性纖維表達(dá)增高,以及TH免疫反應(yīng)陽(yáng)性表達(dá)在黑質(zhì)致密區(qū)傷后上升,而以之相關(guān)部位ChAT陽(yáng)性表達(dá)下降,可能是由于腦損傷后多巴胺能神經(jīng)興奮,引起TH神經(jīng)元合成及釋放增加,中樞膽堿能神經(jīng)和多巴胺能神經(jīng)二者之間可能通過(guò)纖維與胞體或胞體與胞體進(jìn)行突觸聯(lián)系,相互調(diào)節(jié),并具有異同作用,通過(guò)多巴胺神經(jīng)抑制乙酰膽堿能神經(jīng)元釋放乙酰膽堿。
[Abstract]:Objective: To establish a rat model of multiple cerebral concussion (MCC) and observe the changes of dopaminergic neurons, cholinergic neurons and nerve fibers in the brain of MCC rats. METHODS: Thirty-six MCC rats were randomly divided into 6 triple concussion groups (n=6) and a normal control group (n=6) on 1, 2, 4, 8, 16 and 24 days. The dense areas of substantia nigra and anterior cingulate were detected by double-labeled S-P immunohistochemical method with anti-TH and anti-CHAT antibodies. The expression of dopaminergic and cholinergic nerves in the cortex, caudate putamen nucleus, nucleus accumbens, prefrontal cortex, basal forebrain, brainstem reticular structure, Meynert basal nucleus and the relationship between them were studied. Dopaminergic and cholinergic nerves in the basal ganglia of Meynert were counted and the optical density of TH and CHAT positive cells and fibers in these areas were analyzed by image analysis system. Compared with the control group, there was a significant difference (p < 0.05) in 2 days group, 4 days group, 8 days group and 16 days group, and the positive expression of TH cells in substantia nigra compact area was basically restored in 24 days group. The optical density of TH-positive fibers in anterior cingulate cortex, caudate putamen nucleus, nucleus accumbens, prefrontal cortex and medial septal nucleus, perpendicular branch of oblique zone, and horizontal branch in rats were analyzed. Compared with the control group, the TH-positive fibers in each injury group increased, the reaction in 8 days group reached the peak, and the reaction in 24 days group basically returned to normal. There were significant differences in the number of CHAT positive cells in different brain regions after MCC: brainstem reticular structure, medial septal nucleus (MS), diagonal vertical branch (VDB), left and right diagonal horizontal branch (HDB) ChAT positive cells. Compared with the control group, the medial septal nucleus (MS) was significantly different (P The number of ChAT positive cells in the basal ganglia decreased significantly one day and two days after injury, and then gradually increased to normal 24 days after injury. Compared with the control group, the medial septal nucleus (MS) was significantly higher at 16 days after MCC (P < 0.05). The vertical branch of left and right diagonal band (VDB), medial septal nucleus and horizontal branch of right diagonal band (HDB) were significantly lower at 8 days, 16 days and 24 days after MCC. The medial septal nucleus, the vertical branch of left and right diagonal band (VDB) did not return to normal until 24 days after injury. The horizontal branch of left diagonal band (HDB) 8 days, the horizontal branch of right diagonal band (HDB) 24 days compared with the control group were statistically significant (P < 0.05). There was no significant difference between the left and right sides of the basal nucleus of Meynert (P > 0.05). (6) Optical density values of ChAT positive cells and TH positive fibers in brainstem reticular structure (BRF) after MCC: ChAT immunoreactivity in each group of brainstem reticular structure also showed a downward trend, and increased slightly after 4 days, 8 days. Compared with the control group, there were significant differences in pontine nucleus level from 1 day to 24 days (P < 0.05), in trapezoidal nucleus at 1 day, 2 days, 8 days and 16 days after injury (P < 0.05), and in olivary nucleus level at 1 day, 2 days and 16 days (P < 0.05). TH-positive fibers showed an upward trend from 1D to 24D after MCC, reaching a peak at 8D. Compared with the control group, the pontine nucleus at 8d, the trapezoidal nucleus at 16d and the olivary nucleus at 8D had statistical significance (p < 0.05). CONCLUSION: 1. Triple concussion rat model was successfully established by using metal pendulum mechanical percussion device: 2. TH positive expression in putamen nucleus, nucleus accumbens, substantia nigra dense area, medial septal nucleus, diagonal zone and brain stem nucleus increased in varying degrees. The number and activity of cholinergic neurons in basal forebrain and brainstem reticular structure of MCC rats decreased at 8 days and 16 days after injury. 4. After triple concussion, the number and activity of cholinergic neurons in the brain of MCC rats decreased. The increased expression of TH-positive fibers in the cortex, basal forebrain and brainstem nucleus, and the increased expression of TH-immunoreactive fibers in the substantia nigra dense area were observed, whereas the decrease of ChAT-positive expression in the related areas might be due to the increased synthesis and release of TH neurons after brain injury. Additionally, the central cholinergic nerve and the dopaminergic nerve may have synaptic connections with the cell body or the cell body through the fibers, and they may regulate each other, and have similar and different effects. The dopaminergic nerve inhibits the release of acetylcholine from the acetylcholinergic neurons through the dopaminergic nerve.
【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】：2008
【分類(lèi)號(hào)】：R651.15;D919
本文編號(hào)：2189436
[Abstract]:Objective: To establish a rat model of multiple cerebral concussion (MCC) and observe the changes of dopaminergic neurons, cholinergic neurons and nerve fibers in the brain of MCC rats. METHODS: Thirty-six MCC rats were randomly divided into 6 triple concussion groups (n=6) and a normal control group (n=6) on 1, 2, 4, 8, 16 and 24 days. The dense areas of substantia nigra and anterior cingulate were detected by double-labeled S-P immunohistochemical method with anti-TH and anti-CHAT antibodies. The expression of dopaminergic and cholinergic nerves in the cortex, caudate putamen nucleus, nucleus accumbens, prefrontal cortex, basal forebrain, brainstem reticular structure, Meynert basal nucleus and the relationship between them were studied. Dopaminergic and cholinergic nerves in the basal ganglia of Meynert were counted and the optical density of TH and CHAT positive cells and fibers in these areas were analyzed by image analysis system. Compared with the control group, there was a significant difference (p < 0.05) in 2 days group, 4 days group, 8 days group and 16 days group, and the positive expression of TH cells in substantia nigra compact area was basically restored in 24 days group. The optical density of TH-positive fibers in anterior cingulate cortex, caudate putamen nucleus, nucleus accumbens, prefrontal cortex and medial septal nucleus, perpendicular branch of oblique zone, and horizontal branch in rats were analyzed. Compared with the control group, the TH-positive fibers in each injury group increased, the reaction in 8 days group reached the peak, and the reaction in 24 days group basically returned to normal. There were significant differences in the number of CHAT positive cells in different brain regions after MCC: brainstem reticular structure, medial septal nucleus (MS), diagonal vertical branch (VDB), left and right diagonal horizontal branch (HDB) ChAT positive cells. Compared with the control group, the medial septal nucleus (MS) was significantly different (P The number of ChAT positive cells in the basal ganglia decreased significantly one day and two days after injury, and then gradually increased to normal 24 days after injury. Compared with the control group, the medial septal nucleus (MS) was significantly higher at 16 days after MCC (P < 0.05). The vertical branch of left and right diagonal band (VDB), medial septal nucleus and horizontal branch of right diagonal band (HDB) were significantly lower at 8 days, 16 days and 24 days after MCC. The medial septal nucleus, the vertical branch of left and right diagonal band (VDB) did not return to normal until 24 days after injury. The horizontal branch of left diagonal band (HDB) 8 days, the horizontal branch of right diagonal band (HDB) 24 days compared with the control group were statistically significant (P < 0.05). There was no significant difference between the left and right sides of the basal nucleus of Meynert (P > 0.05). (6) Optical density values of ChAT positive cells and TH positive fibers in brainstem reticular structure (BRF) after MCC: ChAT immunoreactivity in each group of brainstem reticular structure also showed a downward trend, and increased slightly after 4 days, 8 days. Compared with the control group, there were significant differences in pontine nucleus level from 1 day to 24 days (P < 0.05), in trapezoidal nucleus at 1 day, 2 days, 8 days and 16 days after injury (P < 0.05), and in olivary nucleus level at 1 day, 2 days and 16 days (P < 0.05). TH-positive fibers showed an upward trend from 1D to 24D after MCC, reaching a peak at 8D. Compared with the control group, the pontine nucleus at 8d, the trapezoidal nucleus at 16d and the olivary nucleus at 8D had statistical significance (p < 0.05). CONCLUSION: 1. Triple concussion rat model was successfully established by using metal pendulum mechanical percussion device: 2. TH positive expression in putamen nucleus, nucleus accumbens, substantia nigra dense area, medial septal nucleus, diagonal zone and brain stem nucleus increased in varying degrees. The number and activity of cholinergic neurons in basal forebrain and brainstem reticular structure of MCC rats decreased at 8 days and 16 days after injury. 4. After triple concussion, the number and activity of cholinergic neurons in the brain of MCC rats decreased. The increased expression of TH-positive fibers in the cortex, basal forebrain and brainstem nucleus, and the increased expression of TH-immunoreactive fibers in the substantia nigra dense area were observed, whereas the decrease of ChAT-positive expression in the related areas might be due to the increased synthesis and release of TH neurons after brain injury. Additionally, the central cholinergic nerve and the dopaminergic nerve may have synaptic connections with the cell body or the cell body through the fibers, and they may regulate each other, and have similar and different effects. The dopaminergic nerve inhibits the release of acetylcholine from the acetylcholinergic neurons through the dopaminergic nerve.
【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】：2008
【分類(lèi)號(hào)】：R651.15;D919
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