Wang Hong, Shao Yihui, Zhang Saisai, Xie Anqi, Ye Yanna, Shi Lihua, Jin Leigang, Pan Xuebo, Lin Zhuofeng, Li Xiaokun, Yang Shulin
Department of Bioengineering, School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, China.
Department of Pharmacy, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325035, China.
Acta Biochim Biophys Sin (Shanghai). 2017 Jun 1;49(6):541-549. doi: 10.1093/abbs/gmx040.
Chemokine C-X-C ligand 16 (CXCL16), a single-pass Type I membrane protein belonging to the CXC chemokine family, is related to the inflammatory response in liver injury. In present study, we investigated the pathophysiological role of CXCL16, a unique membrane-bound chemokine, in acetaminophen (APAP)-induced hepatotoxicity in mice. Mice were injected with APAP, and blood and tissue samples were harvested at different time points. The serum high-mobility group box 1 and CXCL16 levels were quantified by sandwich immunoassays. The liver tissue sections were stained with hematoxylin-eosin or with dihydroethidium staining. The expressions of CXCL16 and other cytokines were examined by real-time polymerase chain reaction. Ly6-B, p-jun N-terminal kinase (p-JNK), and JNK expressions were measured by western blot analysis. Intracellular glutathione, reactive oxygen species, and malondialdehyde levels were also measured. APAP overdose increased hepatic CXCL16 mRNA and serum CXCL16 protein levels. CXCL16-deficient mice exhibited significantly less liver injury and hepatic necrosis, as well as a lower mortality than wild-type (WT) mice in response to APAP-overdose treatment. APAP elevated the production of oxidative stress and decreased mitochondrial respiratory chain activation in WT mice, which was strongly reversed in CXCL16-knockout mice. In addition, CXCL16 deficiency inhibited the neutrophil infiltration and the production of proinflammatory cytokines triggered by APAP-overdose treatment. Our study revealed that CXCL16 is a critical regulator of liver immune response to APAP-induced hepatotoxicity, thus providing a potential strategy for the treatment of drug-induced acute liver failure by targeting CXCL16.
趋化因子C-X-C配体16（CXCL16）是一种属于CXC趋化因子家族的单次跨膜I型膜蛋白，与肝损伤中的炎症反应相关。在本研究中，我们调查了独特的膜结合趋化因子CXCL16在对乙酰氨基酚（APAP）诱导的小鼠肝毒性中的病理生理作用。给小鼠注射APAP，并在不同时间点采集血液和组织样本。通过夹心免疫测定法定量血清高迁移率族蛋白B1和CXCL16水平。肝组织切片用苏木精-伊红染色或用二氢乙锭染色。通过实时聚合酶链反应检测CXCL16和其他细胞因子的表达。通过蛋白质印迹分析测量Ly6-B、磷酸化c-Jun氨基末端激酶（p-JNK）和JNK的表达。还测量了细胞内谷胱甘肽、活性氧和丙二醛水平。APAP过量增加了肝脏CXCL16 mRNA和血清CXCL16蛋白水平。与野生型（WT）小鼠相比，CXCL16缺陷型小鼠在接受APAP过量治疗后表现出明显较轻的肝损伤和肝坏死，以及较低的死亡率。APAP增加了WT小鼠的氧化应激产生并降低了线粒体呼吸链激活，而这在CXCL16基因敲除小鼠中得到了强烈逆转。此外，CXCL16缺陷抑制了APAP过量治疗引发的中性粒细胞浸润和促炎细胞因子的产生。我们的研究表明，CXCL16是肝脏对APAP诱导的肝毒性免疫反应的关键调节因子，从而为通过靶向CXCL16治疗药物性急性肝衰竭提供了一种潜在策略。
