Su Li, Du Hongli, Dong Xin, Zhang Xiuhua, Lou Ziyang
School of Pharmacy, Second Military Medical University, 325 Guo-He Road, Shanghai 200433, China.
J Clin Biochem Nutr. 2016 Sep;59(2):86-92. doi: 10.3164/jcbn.15-128. Epub 2016 Jun 10.
Raf-1 kinase inhibitory protein (RKIP) is a critical molecule for cellular responses to stimuli. In this study, we investigated whether RKIP is responsible for neural cell apoptosis induced by oxygen-glucose deprivation (OGD) and explored the role of NF-κB and ERK pathways regulated by RKIP under OGD stimuli. RKIP was overexpressed or knocked down using lentivirus in PC12 cells, which were then challenged by OGD. RKIP overexpression significantly increased the cell viability of OGD cells, and attenuated apoptosis, cell cycle arrest, and reactive oxygen species generation. RKIP knockdown induced reverse effects. Moreover, we found that RKIP interacted with TAK1, NIK, IKK, and Raf-1 and negatively regulated the NF-κB and ERK pathways. RKIP overexpression significantly inhibited IKK, IκBα, and P65 phosphorylation in NF-κB pathway and MEK, ERK, and CREB phosphorylation in ERK pathway, respectively. RKIP knockdown induced reverse effects. Furthermore, a NF-κB inhibitor BAY 11-7082 and a MEK inhibitor U0126 blocked the changes caused by RKIP down-regulation after OGD. In conclusion, these results demonstrate that RKIP plays a key role in neural cell apoptosis caused by OGD partly via regulating NF-κB and ERK pathways. The present study may provide new insights into the role of RKIP in ischemic stroke.
Raf-1激酶抑制蛋白(RKIP)是细胞对刺激作出反应的关键分子。在本研究中,我们调查了RKIP是否与氧糖剥夺(OGD)诱导的神经细胞凋亡有关,并探讨了在OGD刺激下RKIP调节的NF-κB和ERK信号通路的作用。使用慢病毒在PC12细胞中过表达或敲低RKIP,然后用OGD刺激这些细胞。RKIP过表达显著提高了OGD处理细胞的活力,并减轻了细胞凋亡、细胞周期阻滞和活性氧的产生。RKIP敲低则产生相反的效果。此外,我们发现RKIP与TAK1、NIK、IKK和Raf-1相互作用,并对NF-κB和ERK信号通路起负调节作用。RKIP过表达分别显著抑制了NF-κB信号通路中IKK、IκBα和P65的磷酸化以及ERK信号通路中MEK、ERK和CREB的磷酸化。RKIP敲低则产生相反的效果。此外,NF-κB抑制剂BAY 11-7082和MEK抑制剂U0126可阻断OGD后RKIP下调所引起的变化。总之,这些结果表明,RKIP在OGD引起的神经细胞凋亡中起关键作用,部分是通过调节NF-κB和ERK信号通路实现的。本研究可能为RKIP在缺血性卒中中的作用提供新的见解。