Kong Ling Lei, Wang Zhi Yuan, Hu Jin-Feng, Yuan Yu-He, Han Ning, Li Hua, Chen Nai Hong
(a)State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Neuroscience Center, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China; (b)The Key Lab of Drug Metabolism and Pharmacokinetics, Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China.
The Lab of Biopharmaceutics, Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China.
Neurochem Int. 2014 Oct;76:91-8. doi: 10.1016/j.neuint.2014.07.004. Epub 2014 Jul 18.
Chemokine-like factor 1 (CKLF1) is a novel C-C chemokine, and plays important roles in immune response and brain development. In previous study, we have found that the expression of CKLF1 increased after focal cerebral ischemia and inhibition of CKLF1 using antagonist C19 peptide protected against cerebral ischemia. However, few studies have focused on the role of CKLF1 on neuronal apoptosis. The objective of present study is to investigate the role of CKLF1 on neuronal apoptosis by applying anti-CKLF1 antibodies in rat focal cerebral ischemia and reperfusion model. Antibodies against CKLF1 was applied to the right cerebral ventricle immediately after transient middle cerebral artery occlusion (MCAO), and infarct volume, neurological score, glucose metabolism and apoptosis-related protein were measured. Treatment with anti-CKLF1 antibody decreased infarct volume and neurological score, and inhibited neuronal apoptosis in a dose-dependent manner at 24h after reperfusion. Anti-CKLF1 antibody also reduced the level of phosphorylation of Akt (P-Akt), and led to decrease of pro-apoptotic protein Bcl-2 associated X protein (Bax) and increase of anti-apoptotic protein B cell lymphoma-2 protein (Bcl-2) and the ratio of Bcl-2/Bax, and inhibited caspase-3 at last. In addition, positron emission tomography (PET) indicated that anti-CKLF1 antibody increased glucose metabolism in ischemic hemisphere. These results suggest that CKLF1 is associated with neuronal apoptosis after cerebral ischemia and reperfusion. Neutralization of CKLF1 with antibodies shows neuroprotective effects against cerebral ischemia, which may be involved in inhibition of Akt pathway, regulation of apoptosis-related protein expression, and improvement glucose metabolism in ischemic hemisphere. Therefore, CKLF1 may be a novel target for the treatment of stroke.
趋化因子样因子1(CKLF1)是一种新型的C-C趋化因子,在免疫应答和脑发育中发挥重要作用。在先前的研究中,我们发现局灶性脑缺血后CKLF1的表达增加,并且使用拮抗剂C19肽抑制CKLF1可预防脑缺血。然而,很少有研究关注CKLF1在神经元凋亡中的作用。本研究的目的是通过在大鼠局灶性脑缺血再灌注模型中应用抗CKLF1抗体来研究CKLF1在神经元凋亡中的作用。在短暂性大脑中动脉闭塞(MCAO)后立即将抗CKLF1抗体注入右侧脑室,并测量梗死体积、神经功能评分、葡萄糖代谢和凋亡相关蛋白。抗CKLF1抗体治疗可减少梗死体积和神经功能评分,并在再灌注后24小时以剂量依赖方式抑制神经元凋亡。抗CKLF1抗体还降低了Akt的磷酸化水平(P-Akt),导致促凋亡蛋白Bcl-2相关X蛋白(Bax)减少,抗凋亡蛋白B细胞淋巴瘤-2蛋白(Bcl-2)增加以及Bcl-2/Bax比值增加,最终抑制了caspase-3。此外,正电子发射断层扫描(PET)表明抗CKLF1抗体增加了缺血半球的葡萄糖代谢。这些结果表明CKLF1与脑缺血再灌注后的神经元凋亡有关。用抗体中和CKLF1显示出对脑缺血的神经保护作用,这可能涉及抑制Akt通路、调节凋亡相关蛋白表达以及改善缺血半球的葡萄糖代谢。因此,CKLF1可能是治疗中风的新靶点。