论文部分内容阅读
目的探讨细胞外信号调节蛋白激酶(extracellular signal-regulated protein kinase,ERK)1/2和核转录因子-κB(NF-κB)在胆红素诱导的海马神经细胞凋亡过程中的作用。方法将体外培养的海马神经细胞分为处理组(胆红素处理神经元)、抑制组(胆红素+ERK1/2抑制剂PD98059)以及对照组(DMSO)。采用改良噻唑蓝比色法(MTT法)和Hoechst33342DNA染色法,观察各组细胞存活率及形态学特征;免疫细胞化学检测NF-κB蛋白的表达。结果处理组细胞存活率(85.418±1.406)%明显低于对照组(99.990±2.782)%(P<0.01),而高于抑制组(63.951±1.148)%(P<0.01);对照组和抑制组NF-κB蛋白光密度值、阳性细胞率[分别为(0.157±0.037)、(0.986±0.795)%和(0.249±0.059)、(2.622±1.552)%]均明显低于处理组[分别为(0.306±0.072)、(6.882±2.626)%,P<0.01]。结论胆红素可激活原代培养的海马神经细胞ERK/NF-κB信号转导通路;抑制ERK1/2通路可抑制NF-κB活性而加剧胆红素的神经细胞毒性。
Objective To investigate the role of extracellular signal-regulated protein kinase (ERK) 1/2 and NF-κB in bilirubin-induced hippocampal neuronal apoptosis. Methods The hippocampal neurons cultured in vitro were divided into treatment group (bilirubin-treated neurons), inhibition group (bilirubin + ERK1 / 2 inhibitor PD98059) and control group (DMSO). The cell viability and morphological characteristics were observed by modified thiazolyl blue (MTT) assay and Hoechst33342 DNA staining. The expression of NF-κB protein was detected by immunocytochemistry. Results The cell viability (85.418 ± 1.406)% in the treatment group was significantly lower than that in the control group (99.990 ± 2.782)% (P <0.01), but higher than that in the control group (63.951 ± 1.148)% The optical density and the positive rate of NF-κB in group [(0.157 ± 0.037), (0.986 ± 0.795)% and (0.249 ± 0.059), (2.622 ± 1.552)%] were significantly lower than those in the treatment group [ (0.306 ± 0.072), (6.882 ± 2.626)%, P <0.01]. Conclusion Bilirubin can activate ERK / NF-κB signal transduction pathway in primary cultured hippocampal neurons. Inhibition of ERK1 / 2 pathway can inhibit NF-κB activity and aggravate neurotoxicity of bilirubin.