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目的:研究白藜芦醇(resveratrol,Res)预处理对脑缺血再灌注(ischemia reperfusion,I/R)大鼠海马区Wnt/β-catenin信号通路相关蛋白的表达量、星形胶质细胞及神经元形态的影响,探讨白藜芦醇对脑缺血再灌注大鼠海马区神经元损伤的保护作用机制。方法:将SD大鼠60只随机分为四组,每组15只:正常对照组(Control)、假手术组(Sham)、缺血再灌注组(I/R)和白藜芦醇治疗组(Res)。Res组大鼠连续7 d腹腔注射白藜芦醇(每日30 mg/kg,由2%DMSO溶解),Sham组和I/R组注射同等量2%DMSO;7 d后Res组和I/R组采用改良线栓法制备大鼠大脑中动脉阻塞(middle cerebral artery occlusion,MACO)模型,缺血90 min,再灌注24 h。Sham组仅分离暴露一侧颈内动脉,不插入线栓;24 h后对各组大鼠进行神经功能学评分;脑组织TTC染色;尼氏染色;免疫组化检测海马区GSK-3β和星型胶质细胞(GFAP)数量及形态变化;Western Blot分别检测Wnt信号通路相关因子GSK-3β、p-GSK-3β、β-catenin蛋白水平表达量。结果:再灌注24 h后,尼氏染色显示与I/R组相比Res组海马区仅有少量的神经元破碎和消失,形态尚完整(P<0.05),同时行为学评分和梗死面积也低于I/R组(P<0.05)。免疫组化显示,与I/R组相比Res组GSK-3β的表达量明显降低,伴随星形胶质细胞活化状态和数量减少(P<0.05)。Western Blot显示,与免疫组化表达趋势结果一致Res组GSK-3β的的表达量明显低于I/R组,而p-GSK-3β、β-catenin的表达量均增高(P<0.05)。结论:白藜芦醇可能是通过调节Wnt/β-catenin信号通路相关蛋白的表达进而有效的降低大鼠脑缺血再灌注后神经元的损伤。
OBJECTIVE: To investigate the effect of resveratrol (Res) preconditioning on the expression of Wnt / β-catenin signal pathway related proteins in hippocampus of rats with cerebral ischemia-reperfusion (I / R) And neuron morphology, to explore the protective effect of resveratrol on neuronal damage in hippocampus after cerebral ischemia-reperfusion in rats. Methods: Sixty SD rats were randomly divided into four groups (15 rats in each group): control group, sham group, I / R group and resveratrol group (Res). Rats in Res group were injected intraperitoneally with resveratrol (30 mg / kg daily, dissolved in 2% DMSO) for 7 days. Sham group and I / R group received the same amount of 2% DMSO. After 7 days, Res group and I / Rats in R group were given middle cerebral artery occlusion (MACO) model by modified suture method for 90 min ischemia and 24 h reperfusion. Sham group was isolated only exposed side of the internal carotid artery, not plug; 24 h after the neurological score of each group of rats; brain TTC staining; Nissl staining; immunohistochemical detection of GSK-3β and (GFAP) were detected by Western Blot. The expression of GSK-3β, p-GSK-3β and β-catenin in Wnt signaling pathway were detected by Western Blot. Results: After 24 h of reperfusion, Nissl staining showed that only a small number of neurons in Res hippocampus were broken and disappeared in Res group compared with I / R group (P <0.05), and both behavioral score and infarct size Lower than I / R group (P <0.05). Immunohistochemistry showed that the expression of GSK-3β in Res group was significantly lower than that in I / R group, accompanied by decreased activation status and number of astrocytes (P <0.05). Western Blot showed that the expression level of GSK-3β in Res group was significantly lower than that in I / R group and the expression of p-GSK-3β and β-catenin was increased (P <0.05). Conclusion: Resveratrol may reduce the damage of neurons after cerebral ischemia-reperfusion in rats by regulating the expression of Wnt / β-catenin signaling pathway related proteins.