Effects of Buyang Huanwu Decoction on CXCR4-PI3K autophagy signal transduction pathway and Beclin-1 in a rat model of cerebral hemorrhage

doi:10.3969/j.issn.2095-4344.2016.27.006

Abstract

Abstract:

BACKGROUND: Buyang Huanwu Decoction is commonly used in clinical medicine in treatment of intracerebral hemorrhage. Previous studies have been found that it has excellent neuroprotective effect, can efficiently inhibit the apoptosis of nerve cells. Autophagic activity is closely related to apoptosis of nerve cells. CXCR4-PI3K autophagy signaling pathway has been verified in clinic. However, the effect of Buyang Huanwu Decoction is poorly understood. There is no study on Beclin-1 in the neuroprotective studies of Buyang Huanwu Decoction.
OBJECTIVE: To explore the effects of Buyang Huanwu Decoction on CXCR4-PI3K autophagy signaling pathway and Beclin-1 in rats with cerebral hemorrhage and related mechanisms.
METHODS: According to Rosenberg method, a rat model of cerebral hemorrhage was replicated and intragastrically administered Buyang Huanwu Decoction. Western blot assay was used to measure Beclin-1 protein. Immunohistochemical method was utilized to detect the expression of PI3K, AKT, stromal cell derived factor 1 and CXCR-4 protein. TUNEL assay was applied to identify apoptosis.
RESULTS AND CONCLUSION: (1) After administration, Buyang Huanwu Decoction could reduce the number of neuronal apoptosis in rats with intracerebral hemorrhage, up-regulate the expression of Beclin-1, PI3K, AKT, stromal cell derived factor 1, and CXCR-4 protein, and exert neuroprotective effect. (2) Buyang Huanwu Decoction could activate CXCR4-PI3K autophagy signal transduction pathway, both to stimulate autophagy and to regulate autophagy state, can inhibit apoptosis, and exert cerebral protective effect.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

Key words: Models, Animal, Cerebral Hemorrhage, Apoptosis, Immunochemistry

CLC Number:

R318

Qiu Zhi-fu, Wu Xiao-guang, Zu Bing-xue, Shi Zhao-ping, Miao Hui, Li Meng-meng. Effects of Buyang Huanwu Decoction on CXCR4-PI3K autophagy signal transduction pathway and Beclin-1 in a rat model of cerebral hemorrhage[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(27): 3992-3998.

Figures/Tables 2

2.1 实验动物数量分析实验选用大鼠72只，分成4组，实验过程无脱失，全部进入结果分析。 2.2 细胞凋亡数量的变化用TUNEL法来计算脑组织的神经细胞凋亡数目，TUNEL阳性细胞形态学表现为胞体缩小，核固缩，染色质凝聚，呈棕黄色或黄褐色颗粒，出现明显的凋亡特征。假手术组大鼠大脑皮质内可见零星的、散在的、极为少量的TUNEL阳性细胞，TUNEL阳性细胞中大部分为神经元，少部分为胶质细胞和血管内皮细胞。模型组大鼠血肿周围皮质TUNEL阳性细胞较假手术组显著增加，差异有显著性意义(P < 0.05)；补阳还五汤组和银杏叶片组TUNEL阳性细胞较模型组显著减少，差异有显著性意义(P < 0.05)，详见表1。 2.3 PI3K、AKT蛋白阳性表达时吸光度的变化 PI3K蛋白和AKT蛋白均位于胞浆上，阳性表达呈棕黄色的物质沉积。与假手术组比较，模型组PI3K、AKT蛋白阳性表达显著增高，差异有显著性意义(P < 0.05)；与模型组比较，补阳还五汤组和银杏叶片组PI3K、AKT蛋白阳性表达显著增高，差异有显著性意义(P < 0.05)，详见表2及图1，2。 2.4 Beclin-1的表达变化 Beclin-1条带结果显示，各组中的Beclin-1蛋白表达水平差异均有显著性意义(P < 0.05)。与假手术组相比，模型组的Beclin-1蛋白表达显著升高(P < 0.05)；与模型组相比，补阳还五汤组、银杏叶片组的Beclin-1蛋白表达均显著降低(P < 0.05)，详见表3。 2.5 基质细胞衍生因子1α、CXCR4阳性细胞表达变化基质细胞衍生因子1α阳性细胞表达情况：假手术组中可见0-1个阳性表达，模型组及补阳还五汤组阳性细胞均高于假手术组，且补阳还五汤组阳性细胞数目高于模型组，差异有显著性意义(P < 0.05)，详见表4。 CXCR4阳性细胞表达情况：假手术组可见0-1个阳性表达，模型组及补阳还五汤组阳性细胞均高于假手术组，且补阳还五汤组阳性细胞数目高于模型组，差异有显著性意义(P < 0.05)，详见表4。"

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