Yin Xiao-Ping, Zhou Jun, Wu Dan, Chen Zhi-Ying, Bao Bing
Department of Neurology, Second Affiliated Hospital of Nanchang University Nanchang, China ; Department of Neurology, Affiliated Hospital of Jiujiang University Jiujiang, China.
Department of Neurology, Second Affiliated Hospital of Nanchang University Nanchang, China.
Int J Clin Exp Pathol. 2015 Sep 1;8(9):10436-43. eCollection 2015.
Previous studies indicate that the Nrf2-ARE signaling pathway plays a neruo-protective role in glia cell, however, the mechanism was also elusive. This study aims to explore the inhibitive function of all-trans-retinoic (ATRA) on Nrf2-ARE pathway in intracerebral hemorrhage (ICH), and investigate the mechanism. In this study, the femoral artery injection method was employed to establish ICH model. The model rats were randomly divided into four groups, including Sham group, ICH group, ATRA group and DMSO group. The neurological scores were evaluated for the four groups at different time points. Hematoxylin-Eosin staining was used to stain the CD11b positive glia cells. Double immunofluorescence staining method was utilized to observe the co-expression of HO-1, NF-κB, Nrf2 and TNF-α and CD11b marker in glia cells. Western blot assay was used to detect the Nrf2 protein (total and binding Nrf2), HO-1, NF-κB and TNF-α proteins in every group. The results indicated that neurologiclal scores were significantly decreased in ATRA group compared to ICH gorup (P < 0.05). The glia cells were significantly activated and accumulated in ICH rats. ATRA significantly decreased co-expression of Nrf2, HO-1 and CD11b, and increased co-expression of NF-κB, TNF-α and CD11b of glia cells. ATRA significantly decreased total Nrf2 expression and increased binding Nrf2 expression in ATRA group compared to ICH group (P < 0.05). ATRA decreased anti-oxygen protein Nrf2 and HO-1, and increases inflammatory factors NF-κB and TNF-α. In conclusion, the application of ATRA could inhibit the neuro-protective function effectively by blocking the Nrf2-ARE pathway in glia cells.
以往研究表明,Nrf2-ARE信号通路在胶质细胞中发挥神经保护作用,然而其机制尚不清楚。本研究旨在探讨全反式维甲酸(ATRA)对脑出血(ICH)中Nrf2-ARE通路的抑制作用,并研究其机制。本研究采用股动脉注射法建立ICH模型。将模型大鼠随机分为四组,包括假手术组、ICH组、ATRA组和二甲基亚砜(DMSO)组。在不同时间点对四组大鼠进行神经功能评分。采用苏木精-伊红染色法对CD11b阳性胶质细胞进行染色。利用双重免疫荧光染色法观察胶质细胞中血红素加氧酶-1(HO-1)、核因子κB(NF-κB)、Nrf2和肿瘤坏死因子-α(TNF-α)与CD11b标记物的共表达情况。采用蛋白质免疫印迹法检测各组中Nrf2蛋白(总Nrf2和结合型Nrf2)、HO-1、NF-κB和TNF-α蛋白。结果表明,与ICH组相比,ATRA组神经功能评分显著降低(P<0.05)。ICH大鼠的胶质细胞显著激活并聚集。ATRA显著降低了胶质细胞中Nrf2、HO-1与CD11b的共表达,增加了NF-κB、TNF-α与CD11b的共表达。与ICH组相比,ATRA组总Nrf2表达显著降低,结合型Nrf2表达增加(P<0.05)。ATRA降低了抗氧化蛋白Nrf2和HO-1的表达,增加了炎症因子NF-κB和TNF-α的表达。综上所述,应用ATRA可通过阻断胶质细胞中的Nrf2-ARE通路有效抑制神经保护作用。