卵泡抑素、激活素A与骨形成蛋白-4在缺血缺氧性脑损伤大鼠脑中的表达

发布时间：2019-05-20 00:21

【摘要】：目的本实验探讨卵泡抑素、激活素A与骨形成蛋白-4在正常大鼠和缺血缺氧性脑损伤大鼠脑组织皮层和海马区的表达规律及意义。综合卵泡抑素与两者的作用机制，找到二者信号的交叉点，为临床指导神经重建提供实验依据。方法将同期受孕60只SD（Sprague dawley）大鼠分为正常组和模型组各30只，每组按照胎鼠发育时间随机分为胚胎期8.5天、胚胎期13天、胚胎期18天，出生后3天、出生后7天、出生后30天6个亚组（即E8.5组，E13组，E18组，p3组，P7组，P30组六个亚组），胚胎期和出生后分别采用不同造模方法建立脑缺血缺氧模型，应用SABC免疫组化方法观察检测生长发育不同时期正常大鼠和脑缺血缺氧后大鼠卵泡抑素（Follistatin，FS），，激活素A（Activin A，ACT A)与骨形成蛋白-4（Bone morphogenic protein-4，BMP-4）蛋白的表达情况。通过RT-PCR方法，检测生长发育不同时期正常大鼠和脑缺血缺氧后大鼠卵泡抑素、激活素A与骨形成蛋白-4基因的表达情况。结果 HE染色结果显示缺血缺氧后的脑组织神经元坏死，部分细胞核核固缩深染，局限性神经元数目减少，神经纤维排列紊乱，呈急性缺血改变。神经功能检测发现神经评分随着大鼠年龄的增长而增高（P0.01），即年龄越大神经损伤越加重。通过免疫组织化学方法显示正常组和模型组随着大鼠发育的不同时期脑组织皮层和海马区的卵泡抑素和激活素A表达量逐渐减少（P0.01），BMP-4在胚胎期几乎不表达，出生后30天呈现低表达（P0.01）。RT-PCR方法检测显示缺血缺氧后模型组卵泡抑素、激活素A与骨形成蛋白-4的表达量基本都略高于正常组（P0.01），尤其以卵泡抑素和激活素A的表达增高最为显著，海马区三者的表达不如皮层区增高明显。结论大鼠脑缺血缺氧模型制造成功。卵泡抑素、激活素A与骨形成蛋白-4的表达与生长发育和年龄密切相关，随着大鼠年龄的增长，卵泡抑素和激活素A的表达逐渐减少，而骨形成蛋白-4的表达则在成年后明显。脑缺血缺氧损伤可诱导卵泡抑素、激活素A和骨形成蛋白-4的高表达，是神经损伤的保护性因子，且卵泡抑素和激活素A是生长发育早期的保护性因子。但是BMP-4的表达增高不如卵泡抑素和激活素A明显，推测卵泡抑素的主要功能可能是作为激活素A的抑制剂而不是骨形成蛋白-4的配体来调控神经发育。
[Abstract]:Objective to investigate the expression and significance of follicular somatostatin, activin A and bone morphogenetic protein-4 in cerebral cortex and hippocampus of normal rats and rats with ischemic and anoxic brain injury. To find the intersection of the two signals, and to provide experimental basis for clinical guidance of nerve reconstruction. Methods 60 SD (Sprague dawley) rats were divided into normal group (n = 30) and model group (n = 30). According to the development time of fetal rats, each group was randomly divided into embryonic stage (8.5 days), embryonic stage (13 days), embryonic stage (18 days) and postbirth (3 days). Cerebral ischemia and hypoxia models were established in 6 subgroup groups (E8.5 group, E13 group, E18 group, p3 group, P7 group, P30 group) 7 days after birth and 30 days after birth. SABC Immunohistochemical method was used to detect Follistatin,FS, Activin A and bone morphogenetic protein-4 (Bone morphogenic protein-4, in normal rats and rats after cerebral ischemia and hypoxia at different stages of growth and development. The expression of BMP-4) protein. The expression of follicle somatostatin, activin A and bone morphogenetic protein-4 gene in normal rats and rats after cerebral ischemia and hypoxia at different stages of growth and development were detected by RT-PCR. Results the results of HE staining showed that after ischemia and hypoxia, the neurons of brain tissue were necrotic, some of the nucleus was stained deeply, the number of localized neurons decreased, and the arrangement of nerve fibers was disordered, showing acute ischemic changes. Nerve function test showed that the nerve score increased with the increase of age (P 0.01), that is to say, the older the age, the more serious the nerve injury. The expression of follicle somatostatin and activin A in cerebral cortex and hippocampus of normal group and model group decreased gradually with the development of rats at different stages of development (P01), but BMP-4 was almost not expressed in embryonic stage. The expression of follicle somatostatin, activin A and bone morphogenetic protein-4 in the model group was slightly higher than that in the normal group after ischemia and hypoxia (P01). The expression of follicle somatostatin, activin A and bone morphogenetic protein-4 in the model group was slightly higher than that in the normal group (P 0.01). In particular, the expression of follicle somatostatin and activin A was the most significant, and the expression of follicle somatostatin and activin A in hippocampus was not as high as that in cortical area. Conclusion the rat model of cerebral ischemia and hypoxia is successful. The expression of follicle somatostatin, activin A and bone morphogenetic protein-4 was closely related to growth, development and age. With the increase of age, the expression of follicle somatostatin and activin A decreased gradually. The expression of bone morphogenetic protein-4 was obvious in adulthood. Cerebral ischemia and hypoxia injury can induce the high expression of follicle somatostatin, activin A and bone morphogenetic protein-4, which is the protective factor of nerve injury, and follicle somatostatin and activin A are protective factors in the early stage of growth and development. However, the increased expression of BMP-4 is not as obvious as that of follicular somatostatin and activin A. it is speculated that the main function of folliculatostatin may be to regulate nerve development as an inhibitor of activin A rather than as a ligand of bone morphogenetic protein-4.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743

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