【摘要】： 第一部分皮質(zhì)脊髓束在大鼠脊髓內(nèi)的精確定位 目的：確定大鼠皮質(zhì)脊髓束在延髓和脊髓白質(zhì)內(nèi)的走行和精確定位。方法：選用成年SD大鼠，采用Luxol fast blue(LFB、)染色法和protein kinase Cγ(PKCγ)特異性抗體免疫組織化學(xué)染色法在冠狀面、矢狀面、橫斷面三種不同切面上對(duì)正常大鼠皮質(zhì)脊髓束進(jìn)行解剖定位。結(jié)果：在延髓錐體中，LFB染色標(biāo)記的深藍(lán)色的有髓纖維經(jīng)錐體交叉至脊髓頸、胸、腰、骶段后索腹側(cè)，與周圍淡染的纖維易于區(qū)別；PKCγ陽(yáng)性反應(yīng)產(chǎn)物呈棕色，分布于延髓腹側(cè)面的錐體中，在延髓錐體下部絕大部分交叉到背側(cè)面，在脊髓白質(zhì)內(nèi)沿后索腹側(cè)下行，至脊髓的骶尾段。同時(shí)延髓和脊髓背角神經(jīng)元中也有PKCγ免疫陽(yáng)性物質(zhì)存在，在脊髓前索和外側(cè)索未見(jiàn)有PKCγ陽(yáng)性反應(yīng)產(chǎn)物。LFB陽(yáng)性纖維和PKCγ免疫陽(yáng)性反應(yīng)產(chǎn)物在延髓錐體和脊髓后索內(nèi)的定位分布與大鼠皮質(zhì)脊髓束在延髓和脊髓中的位置和走行相一致。結(jié)論：運(yùn)用LFB染色和PKCγ特異性抗體免疫組織化學(xué)染色可以清晰對(duì)正常大鼠皮質(zhì)脊髓束進(jìn)行精確解剖定位，為進(jìn)一步研究皮質(zhì)脊髓束的損傷和功能重建提供形態(tài)學(xué)基礎(chǔ)。 第二部分大鼠皮質(zhì)脊髓束半橫段損傷模型的建立 目的：成功制作大鼠皮質(zhì)脊髓束半橫斷損傷模型。方法：選擇性切割大鼠延髓左側(cè)錐體，建立大鼠皮質(zhì)脊髓束半橫斷損傷模型，采用斜板實(shí)驗(yàn)進(jìn)行行為學(xué)檢測(cè)，運(yùn)用LFB染色法、生物素化葡聚糖胺(BDA)神經(jīng)示蹤法和PKCγ免疫組織化學(xué)染色法對(duì)模型進(jìn)行形態(tài)學(xué)評(píng)判。結(jié)果：皮質(zhì)脊髓束損傷組大鼠術(shù)后右側(cè)前后肢癱瘓，，斜板實(shí)驗(yàn)顯示運(yùn)動(dòng)功能受限；LFB染色可以看出損傷部位左側(cè)皮質(zhì)脊髓束無(wú)Luxol Fast Blue濃染神經(jīng)纖維束；BDA神經(jīng)示蹤法結(jié)果顯示在錐體交叉平面，僅見(jiàn)一束B(niǎo)DA陽(yáng)性反應(yīng)產(chǎn)物經(jīng)右側(cè)交叉至左側(cè)，在脊髓后索的左側(cè)半下行至骶尾段，而在損傷平面以下，未見(jiàn)有BDA陽(yáng)性反應(yīng)產(chǎn)物經(jīng)左側(cè)錐體交叉至右側(cè)，在整個(gè)脊髓后索的右側(cè)半未見(jiàn)有BDA陽(yáng)性反應(yīng)產(chǎn)物。PKCγ免疫組織化學(xué)染色結(jié)果發(fā)現(xiàn)延髓錐體右側(cè)半有一束PKCγ陽(yáng)性反應(yīng)產(chǎn)物經(jīng)右側(cè)交叉至左側(cè)，脊髓后索腹側(cè)部左側(cè)半有PKCγ陽(yáng)性反應(yīng)產(chǎn)物，而脊髓后索腹側(cè)部右側(cè)半未見(jiàn)有PKCγ陽(yáng)性反應(yīng)產(chǎn)物。結(jié)論：選擇在延髓切斷一側(cè)錐體制作皮質(zhì)脊髓束半橫斷損傷模型，定位準(zhǔn)確，重復(fù)性好，結(jié)果可靠，是研究皮質(zhì)脊髓束的可塑性和神經(jīng)軸突再生的理想動(dòng)物模型。BDA神經(jīng)束路示蹤法和PKCγ免疫組織化學(xué)染色法可作為皮質(zhì)脊髓束半橫斷損傷模型的形態(tài)學(xué)指標(biāo)。 第三部分大鼠皮質(zhì)脊髓束半橫斷損傷后皮質(zhì)脊髓束超微結(jié)構(gòu)的變化 目的：觀察皮質(zhì)脊髓束半橫斷損傷后皮質(zhì)脊髓束超微結(jié)構(gòu)的變化。方法：選擇性切割大鼠延髓左側(cè)錐體，建立皮質(zhì)脊髓束半橫斷損傷模型，模型制作后4d、14d、28d，采用透射電鏡觀察受損皮質(zhì)脊髓束超微結(jié)構(gòu)的變化。結(jié)果：皮質(zhì)脊髓束半橫斷損傷后，受損皮質(zhì)脊髓束髓鞘和軸索發(fā)生腫脹，形態(tài)不規(guī)則。隨著時(shí)間的延長(zhǎng)，潰變進(jìn)行性加重，受損皮質(zhì)脊髓束主要表現(xiàn)為髓鞘破壞、溶解及軸索脫髓鞘病變、胞漿濃縮、細(xì)胞器增多及空泡樣變性。脊髓頸、胸、腰段受損皮質(zhì)脊髓束的潰變比損傷部位嚴(yán)重。結(jié)論：皮質(zhì)脊髓束半橫斷損傷后，受損皮質(zhì)脊髓束中的髓鞘和軸索發(fā)生進(jìn)行性潰變。
[Abstract]:The first part is the precise location of corticospinal tract in the spinal cord of rats.
Objective: To determine the course and precise location of the corticospinal tract in the medulla oblongata and white matter of the spinal cord in rats.Methods: Adult SD rats were selected and the cortex of normal rats was treated with Luxol fast blue (LFB,) staining and protein kinase C gamma (PKC gamma) specific antibody immunohistochemical staining on coronal, sagittal and transverse sections. Results: In the medullary pyramid, the dark blue myelinated fibers labeled by LFB staining crossed through the pyramid to the cervical, thoracic, lumbar, and ventral sacral posterior funiculus, easily distinguished from the lightly stained fibers around them; the PKC-gamma positive products were brown, distributed in the pyramidal body of the ventral side of the medulla oblongata, and mostly intersected in the lower part of the medullary pyramid. PKC-gamma immunoreactive substances were also present in the medulla oblongata and dorsal horn neurons. No PKC-gamma immunoreactive products were found in the anterior and lateral spinal cord. LFB-positive fibers and PKC-gamma immunoreactive products were identified in the medulla oblongata and the posterior spinal cord. CONCLUSION: LFB staining and PKC gamma specific antibody immunohistochemical staining can be used to precisely locate the corticospinal tract in normal rats and provide morphological basis for further study of corticospinal tract injury and functional reconstruction.
The second part is the establishment of a semi transverse injury model of corticospinal tract in rats.
Objective: To establish a rat model of corticospinal tract hemisection injury. Methods: The left pyramidal body of medulla oblongata was cut selectively to establish a rat model of corticospinal tract hemisection injury. Results: In the corticospinal tract injury group, the right anterior and posterior limbs were paralyzed, and the oblique plate test showed that the motor function was limited; LFB staining showed that there was no Luxol Fast Blue densely stained nerve fiber bundle in the left corticospinal tract; BDA nerve tracing showed only one nerve bundle in the pyramidal cross plane. BDA positive products crossed to the left side through the right side, and then to the sacrococcygeal segment in the left half of the posterior funiculus of the spinal cord. No BDA positive products crossed to the right side through the left pyramid below the injury level. No BDA positive products were found in the right half of the posterior funiculus of the whole spinal cord. There was a bunch of PKC-gamma positive products crossed from right side to left side in the right half, and PKC-gamma positive products were found in the left side of the ventral part of the posterior funiculus, but no PKC-gamma positive products were found in the right side of the ventral part of the posterior funiculus. BDA tract tracing method and PKC-gamma immunohistochemical staining method can be used as morphological indexes for the model of corticospinal tract hemisection injury.
The third part is the ultrastructural changes of corticospinal tract after corticospinal tract hemisection in rats.
Objective:To observe the ultrastructural changes of the corticospinal tract after hemisection of the corticospinal tract.Methods:The left pyramid of the medulla oblongata was cut selectively to establish a model of hemisection of the corticospinal tract in rats. After transection injury, the myelin sheath and axon of the injured corticospinal tract were swollen and irregular in shape. With the prolongation of time, the degeneration of the injured corticospinal tract was progressively aggravated. The main manifestations of the injured corticospinal tract were myelin sheath destruction, lysis and axonal demyelination, cytoplasmic concentration, increased organelles and vacuole-like degeneration. Conclusion: The myelin sheath and axons in the injured corticospinal tract degenerate progressively after hemisection of corticospinal tract.
【學(xué)位授予單位】：南通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R322

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