大鼠皮质脊髓束的应用解剖学研究

发布时间：2018-08-22 12:54

【摘要】： 第一部分皮质脊髓束在大鼠脊髓内的精确定位目的：确定大鼠皮质脊髓束在延髓和脊髓白质内的走行和精确定位。方法：选用成年SD大鼠，采用Luxol fast blue(LFB、)染色法和protein kinase Cγ(PKCγ)特异性抗体免疫组织化学染色法在冠状面、矢状面、横断面三种不同切面上对正常大鼠皮质脊髓束进行解剖定位。结果：在延髓锥体中，LFB染色标记的深蓝色的有髓纤维经锥体交叉至脊髓颈、胸、腰、骶段后索腹侧，与周围淡染的纤维易于区别；PKCγ阳性反应产物呈棕色，分布于延髓腹侧面的锥体中，在延髓锥体下部绝大部分交叉到背侧面，在脊髓白质内沿后索腹侧下行，至脊髓的骶尾段。同时延髓和脊髓背角神经元中也有PKCγ免疫阳性物质存在，在脊髓前索和外侧索未见有PKCγ阳性反应产物。LFB阳性纤维和PKCγ免疫阳性反应产物在延髓锥体和脊髓后索内的定位分布与大鼠皮质脊髓束在延髓和脊髓中的位置和走行相一致。结论：运用LFB染色和PKCγ特异性抗体免疫组织化学染色可以清晰对正常大鼠皮质脊髓束进行精确解剖定位，为进一步研究皮质脊髓束的损伤和功能重建提供形态学基础。第二部分大鼠皮质脊髓束半横段损伤模型的建立目的：成功制作大鼠皮质脊髓束半横断损伤模型。方法：选择性切割大鼠延髓左侧锥体，建立大鼠皮质脊髓束半横断损伤模型，采用斜板实验进行行为学检测，运用LFB染色法、生物素化葡聚糖胺(BDA)神经示踪法和PKCγ免疫组织化学染色法对模型进行形态学评判。结果：皮质脊髓束损伤组大鼠术后右侧前后肢瘫痪，，斜板实验显示运动功能受限；LFB染色可以看出损伤部位左侧皮质脊髓束无Luxol Fast Blue浓染神经纤维束；BDA神经示踪法结果显示在锥体交叉平面，仅见一束BDA阳性反应产物经右侧交叉至左侧，在脊髓后索的左侧半下行至骶尾段，而在损伤平面以下，未见有BDA阳性反应产物经左侧锥体交叉至右侧，在整个脊髓后索的右侧半未见有BDA阳性反应产物。PKCγ免疫组织化学染色结果发现延髓锥体右侧半有一束PKCγ阳性反应产物经右侧交叉至左侧，脊髓后索腹侧部左侧半有PKCγ阳性反应产物，而脊髓后索腹侧部右侧半未见有PKCγ阳性反应产物。结论：选择在延髓切断一侧锥体制作皮质脊髓束半横断损伤模型，定位准确，重复性好，结果可靠，是研究皮质脊髓束的可塑性和神经轴突再生的理想动物模型。BDA神经束路示踪法和PKCγ免疫组织化学染色法可作为皮质脊髓束半横断损伤模型的形态学指标。第三部分大鼠皮质脊髓束半横断损伤后皮质脊髓束超微结构的变化目的：观察皮质脊髓束半横断损伤后皮质脊髓束超微结构的变化。方法：选择性切割大鼠延髓左侧锥体，建立皮质脊髓束半横断损伤模型，模型制作后4d、14d、28d，采用透射电镜观察受损皮质脊髓束超微结构的变化。结果：皮质脊髓束半横断损伤后，受损皮质脊髓束髓鞘和轴索发生肿胀，形态不规则。随着时间的延长，溃变进行性加重，受损皮质脊髓束主要表现为髓鞘破坏、溶解及轴索脱髓鞘病变、胞浆浓缩、细胞器增多及空泡样变性。脊髓颈、胸、腰段受损皮质脊髓束的溃变比损伤部位严重。结论：皮质脊髓束半横断损伤后，受损皮质脊髓束中的髓鞘和轴索发生进行性溃变。
[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.
【学位授予单位】：南通大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R322

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