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目的观察黄芪注射液对缺氧缺血大鼠脑组织神经细胞形态及Bc1-2、Bax蛋白表达的影响,探讨其对脑组织的保护作用及其机制。方法应用右侧颈总动脉结扎、60mL.L-1氧、940mL.L-1氮混合气体处理4h的方法制备缺氧缺血大鼠模型。实验分为假手术组(A组,n=10),模型组(B组,n=20),黄芪注射液组(C组,n=20)。3组大鼠均于脑缺氧缺血24h取脑组织标本,检测Bc1-2、Bax蛋白表达及神经细胞凋亡。结果 1.HE染色:A组幼鼠额叶皮质神经元分布均匀,形态无异常。B组幼鼠额叶皮质神经元排列紊乱,神经元肿胀变性,胞浆空化,部分细胞核呈颗粒样变性,核染色质边集、碎裂和溶解。亦可见胞质浓缩变暗,整个神经细胞体积缩小,胞膜皱缩,呈三角形或不规则形。胞膜与周围分界明显,有的神经元周围有空泡形成。与B组比较,C组幼鼠神经元镜下改变明显减轻。2.尼氏染色:A组幼鼠额叶皮质细胞层次清晰,胞质内尼氏体均匀蓝染。B组幼鼠额叶皮质细胞数减少,神经元排列疏松,皱缩成三角形或锥形,神经元胞体和树突内尼氏体溶解消失。与B组比较,C组幼鼠神经元丢失明显减轻。3.免疫组织化学检测Bcl-2和Bax蛋白表达阳性的细胞数。A组未见Bcl-2和Bax蛋白阳性细胞数增多。B组幼鼠额叶皮层Bc1-2及Bax蛋白阳性细胞数增多(Pa<0.01)。与B组比较,C组幼鼠额叶皮层Bc1-2蛋白阳性细胞数增多,Bax蛋白阳性细胞数降低(P<0.01)。结论黄芪注射液可通过上调缺氧缺血大鼠脑组织Bc1-2蛋白表达,下调Bax蛋白表达,Bc1-2/Bax比值升高,从而抑制神经细胞凋亡,减轻缺氧缺血所致的脑组织损伤。
Objective To observe the effect of astragalus injection on the neuronal morphology and the expression of Bcl-2 and Bax proteins in the brain of hypoxic-ischemic rats and to explore its protective effect on brain and its mechanism. Methods Hypoxic-ischemic rat model was established by ligation of right common carotid artery, 60 mL.L-1 oxygen and 940 mL.L-1 nitrogen mixed gas for 4 h. The experiment was divided into sham operation group (group A, n = 10), model group (group B, n = 20) and Astragalus injection group (group C, n = 20). The brain tissue samples of all three groups were taken at 24 hours after hypoxic-ischemic brain injury to detect the expression of Bc1-2, Bax and the apoptosis of nerve cells. HE staining: A group of young rat frontal cortex neurons evenly distributed, no abnormal morphology. In group B, frontal cortex neurons were disordered, neurons were swollen and degenerated, cytoplasm cavitated, part of the nucleus was granular degeneration, nuclear chromatin margination, fragmentation and lysis. Can also be seen darkening of the cytoplasm concentration, the entire nerve cell volume decreases, cell membrane shrinkage, triangular or irregular shape. Membrane and the surrounding obvious boundaries, some neurons around the formation of vacuoles. Compared with group B, the changes of neurons in group C were significantly reduced. 2. Nissl staining: A group of young rat frontal cortex clear level of cytoplasm Nissl body uniform blue dye. In group B, the number of frontal lobe cortical cells decreased, neurons arranged loosely, and contracted into a triangle or pyramid. The neuronal somatic cells and dendritic Nissl bodies dissolved and disappeared. Compared with group B, the loss of neurons in group C was significantly reduced. Immunohistochemical detection of Bcl-2 and Bax protein positive cells. In group A, there was no increase in the number of Bcl-2 and Bax positive cells. The positive cells of Bcl-2 and Bax in frontal cortex of group B were increased (Pa <0.01). Compared with group B, the number of Bc1-2 positive cells and the number of Bax positive cells in frontal cortex of group C increased (P <0.01). Conclusion Astragalus injection can up-regulate the expression of Bcl-2 protein, decrease the expression of Bax protein and increase the ratio of Bc1-2 / Bax in hypoxic-ischemic brain tissue, thereby inhibiting neuronal apoptosis and alleviating hypoxia-ischemia Brain tissue injury.