【摘要】： 第一部分不同年齡大鼠三叉神經(jīng)節(jié)、頸交感神經(jīng)節(jié)、脊髓背根神經(jīng)節(jié)內(nèi)神經(jīng)前體細(xì)胞的觀察 目的觀察胚胎、新生和成年大鼠三叉神經(jīng)節(jié)、頸交感神經(jīng)節(jié)和脊髓背根神經(jīng)節(jié)內(nèi)是否存在神經(jīng)前體細(xì)胞。方法實驗動物經(jīng)灌流后,取胚胎、新生和成年大鼠三叉神經(jīng)節(jié)、頸交感神經(jīng)節(jié)和脊髓背根神經(jīng)節(jié)行冰凍切片,進(jìn)行p75,Nestin免疫組織化學(xué)和Brdu/ Nestin,Brdu/ p75免疫熒光組織化學(xué)檢測。另取上述神經(jīng)節(jié)進(jìn)行原代培養(yǎng)和傳代培養(yǎng),觀察其體外生長情況。以胎牛血清對傳代培養(yǎng)細(xì)胞進(jìn)行誘導(dǎo)分化。對原代細(xì)胞進(jìn)行Nestin免疫組織化學(xué)和Brdu免疫熒光鑒定;對傳代誘導(dǎo)細(xì)胞進(jìn)行NF、GFAP免疫熒光鑒定。結(jié)果在胚胎、新生、成年大鼠的三叉神經(jīng)節(jié)、脊髓背根神經(jīng)節(jié)內(nèi)均見有Brdu/ Nestin,Brdu/ p75免疫熒光雙標(biāo)細(xì)胞。取自胚胎和新生大鼠三叉神經(jīng)節(jié)和脊髓背根神經(jīng)節(jié)的細(xì)胞經(jīng)原代培養(yǎng),呈現(xiàn)Nestin和Brdu免疫熒光陽性。培養(yǎng)第三代細(xì)胞加入胎牛血清后出現(xiàn)分化,呈現(xiàn)NF、GFAP免疫熒光陽性。結(jié)論:在胚胎、新生、成年大鼠的三叉神經(jīng)節(jié)、脊髓背根神經(jīng)節(jié)和成年大鼠的頸交感神經(jīng)節(jié)中存在有增殖分裂的神經(jīng)前體細(xì)胞,這些細(xì)胞可以分化為神經(jīng)元和神經(jīng)膠質(zhì)細(xì)胞。 第二部分疼痛刺激對成年大鼠脊髓背根神經(jīng)節(jié)內(nèi)神經(jīng)前體細(xì)胞增殖與分化的影響 目的評價疼痛刺激是否會激發(fā)成年大鼠的脊髓背根神經(jīng)節(jié)內(nèi)的神經(jīng)前體細(xì)胞的增殖與分化。方法用1%福爾馬林注射于大鼠左前掌制造疼痛模型,將疼痛模型分為3天組和7天組,各4只,另設(shè)正常對照組4只。3組皆連續(xù)給予Brdu標(biāo)記3天, 3天后將動物分別于不同時間點處死。對正常對照組和模型3天組大鼠的背根節(jié)進(jìn)行Brdu/ Nestin免疫熒光雙標(biāo)檢測,比較疼痛通路上的背根節(jié)內(nèi),與非疼痛通路上的背根節(jié)和正常組的大鼠背根節(jié)內(nèi)Brdu/ Nestin免疫熒光雙標(biāo)細(xì)胞數(shù)目。模型7天組的疼痛通路上背根節(jié)與非疼痛通路上背根節(jié)行BrdU/NF、BrdU/GFAP免疫熒光雙標(biāo)檢測,比較各組雙標(biāo)細(xì)胞數(shù)。結(jié)果3組大鼠各節(jié)段背根節(jié)內(nèi)都可見到Brdu/ Nestin雙標(biāo)細(xì)胞,但疼痛通路上的雙標(biāo)細(xì)胞數(shù)量并未見與非疼痛通路和正常組的雙標(biāo)細(xì)胞數(shù)量有明顯差別。7天組背根節(jié)內(nèi)的BrdU/GFAP雙標(biāo)細(xì)胞數(shù)量多于BrdU/NF雙標(biāo)細(xì)胞。結(jié)論疼痛刺激對成年大鼠背根節(jié)內(nèi)神經(jīng)前體細(xì)胞的增殖作用并不明顯,成年大鼠背根節(jié)內(nèi)增殖的前體細(xì)胞在疼痛刺激的作用下主要分化為神經(jīng)膠質(zhì)細(xì)胞。
[Abstract]:Part one: observation of neural precursor cells in trigeminal ganglion, cervical sympathetic ganglion and dorsal root ganglion of spinal cord in rats of different ages objective to observe the trigeminal ganglion of embryonic, newborn and adult rats, Whether neural progenitor cells exist in cervical sympathetic ganglia and dorsal root ganglion of spinal cord. Methods Frozen sections of trigeminal ganglion, cervical sympathetic ganglia and dorsal root ganglia of spinal cord were obtained from embryonic, newborn and adult rats after perfusion. P75, nestin immunohistochemistry and Brdu/ Nestin, were performed. Brdu/ p75 immunofluorescence histochemistry. In addition, the primary culture and subculture of the above ganglia were performed to observe the growth of the ganglia in vitro. Fetal bovine serum was used to induce the differentiation of subcultured cells. Primary cells were identified by Nestin immunohistochemistry and Brdu immunofluorescence, and subcultured cells were identified by NF,GFAP immunofluorescence. Results Brdu/ Nestin,Brdu/ p75 immunofluorescence double labeled cells were found in trigeminal ganglion and dorsal root ganglia of embryonic, neonatal and adult rats. The cells from trigeminal ganglion and spinal dorsal root ganglion of embryonic and neonatal rats were primarily cultured and showed positive immunofluorescence of Nestin and Brdu. The third generation cells were differentiated with fetal bovine serum and showed NF,GFAP immunofluorescence positive. Conclusion: in trigeminal ganglion of embryonic newborn adult rat dorsal root ganglion of spinal cord and cervical sympathetic ganglia of adult rat there are proliferating and mitotic neural progenitor cells which can differentiate into neurons and glial cells. Part two effect of pain stimulation on proliferation and differentiation of neural precursor cells in spinal dorsal root ganglion of adult rats objective to evaluate whether pain stimulation can stimulate the spinal dorsal root nerve of adult rats Proliferation and differentiation of neural precursor cells. Methods the pain model was made by injecting 1% formalin into the left anterior metacarpal of rats. The pain model was divided into 3-day group and 7-day group with 4 rats in each group, and 4 rats in the normal control group. All of the 3 groups were labeled with Brdu for 3 days. The animals were killed at different time points after 3 days. The dorsal root ganglia of normal control group and model 3 day group were detected by Brdu/ Nestin immunofluorescence double labeling, and the pain pathway in the dorsal root ganglia was compared. The number of Brdu/ Nestin immunofluorescence double labeled cells in dorsal root ganglia with non-pain pathway and normal rats. The dorsal root ganglia on pain pathway and the dorsal root node on non-pain pathway in the 7-day model group were detected by BrdU/NF,BrdU/GFAP immunofluorescence double labeling, and the number of double-labeled cells in each group was compared. Results double labeled Brdu/ Nestin cells were observed in the dorsal root ganglia of the three groups. However, the number of double labeled cells in the pain pathway was not significantly different from that in the non-pain pathway and the normal group. The number of BrdU/GFAP double-labeled cells in the dorsal root ganglion of the 7-day group was higher than that of the BrdU/NF double-labeled cells. Conclusion the effect of pain stimulation on the proliferation of neural progenitor cells in the dorsal root ganglion of adult rats is not obvious. The neural progenitor cells proliferated in the dorsal root ganglion of adult rats are mainly differentiated into glial cells under the action of pain stimulation.
【學(xué)位授予單位】：東南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R329

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