补阳还五汤对脑出血大鼠PI3K/AKT信号通路的影响及其神经保护作用的机制

目的观察并探究补阳还五汤对脑出血大鼠脑组织磷脂酰肌醇3激酶/丝氨酸/苏氨酸蛋白激酶(PI3K/AKT)信号转导通路的影响及其神经保护作用的可能机制。方法 72只SD大鼠随机分为四组:假手术组、模型组、补阳还五汤组、银杏叶片组,每组18只。其中模型组、补阳还五汤组、银杏叶片组采用Rosenberg法制作脑出血大鼠模型。Garcia法检测大鼠神经功能评分,电镜观察神经元线粒体超微结构,western blot法检测磷酸化蛋白激酶(p-AKT)蛋白表达,TUNEL法检测细胞凋亡变化,免疫组化法检测B细胞淋巴瘤基因-2(bcl-2)蛋白、Bcl-2相关X蛋白(bax)的表达,甲酰胺法检测血脑屏障(BBB)通透性,干湿重法检测脑组织含水量。结果与脑出血模型组相比,补阳还五汤治疗明显提高神经功能学评分(P0.05),降低BBB通透性,减少脑组织水含量(P0.05),上调p-AKT、bcl-2蛋白表达,下调bax蛋白表达,减轻线粒体损伤,抑制神经元凋亡。结论补阳还五汤抗脑出血引起脑损伤的作用机制可能与其激活PI3K/AKT信号途径,抑制神经元凋亡,降低BBB通透性,减轻脑水肿有关。

Neuroprotective effect of Buyang Huanwu decoction on the PI3K/AKT signaling pathway in rats with cerebral hemorrhage and possible mechanisms

Objective To investigate the neuroprotective effect of Buyang Huanwu decoction on the phosphatidylinositol 3-kinase/serine/threonine protein kinase (PI3K/AKT) signaling pathway in the brain tissue of rats with cerebral hemorrhage and possible mechanisms.Methods A total of 72 Sprague-Dawley rats were randomly divided into sham-operation group, model group, Buyang Huanwu decoction group, and Ginkgo leaf tablet group, with 18 rats in each group. In the model group, Buyang Huanwu decoction group, and Ginkgo leaf tablet group, the Rosenberg method was used to establish the rat model of cerebral hemorrhage. The Garcia method was used to determine the neurological score, electron microscopy was performed to observe the neuronal mitochondrial ultrastructure, Western blot was used to measure the protein expression of phosphorylated protein kinase (p-AKT), TUNEL was used to measure the changes in cell apoptosis, immunohistochemistry was used to measure the expression of B cell lymphoma-2 protein (bcl-2) and bcl-2-associated X protein (bax), formamide assay was performed to determine the permeability of blood-brain barrier (BBB), and the wet-dry weighting method was used to measure the water content in brain tissue.Results Compared with the cerebral hemorrhage model group, Buyang Huanwu decoction significantly increased neurological score (P<0.05), reduced the permeability of BBB, reduced the water content in brain tissue (P<0.05), upreguated the protein expression of p-AKT and bcl-2, downregulated the protein expression of bax, relieved mitochondrial injury, and inhibited neuronal apoptosis.Conclusions The mechanism of the protective effect of Buyang Huanwu decoction against cerebral hemorrhage caused by brain injury may be related to the activation of the PI3K/AKT signaling pathway, inhibition of neuronal apoptosis, reduction in BBB permeability, and alleviation of cerebral edema.