神经酰胺在Wilson病铜累积诱导肝损伤和肝细胞凋亡中的作用和机制  

Role and mechanism of ceramide in copper accumulation induced liver injury and hepatocyte apoptosis in Wilson’s disease

作　　者：李潇瑾 李艳萌[1] 李振坤 徐安健[1] 杨晓曦 黄坚[1] LI Xiao-jin;LI Yan-meng;LI Zhen-kun(Experimental Center,Beijing Friendship Hospital,Capital Medical University/Beijing Clinical Medicine Institute,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院科研实验中心,北京市临床医学研究所,北京100050

出　　处：《临床和实验医学杂志》2022年第11期1121-1125,共5页Journal of Clinical and Experimental Medicine

基　　金：国家自然科学基金(编号:81602032);北京市优秀人才项目(编号:2016000021469G224)。

摘　　要：目的探讨神经酰胺在威尔逊病(WD)铜累积诱导肝细胞损伤凋亡中的调控作用及其机制。方法对WD小鼠肝组织进行转录组测序并对神经酰胺相关差异基因进行富集分析和反转录实时荧光定量聚合酶链式反应(q RT-PCR)验证。采用Gene MANIA数据库构建神经酰胺相关差异蛋白和凋亡效应蛋白互作网络(PPI)。利用不同浓度高铜溶液处理肝癌细胞,分析胞内铜累积、细胞凋亡率与胞内神经酰胺含量的相关性。利用多球壳菌素抑制神经酰胺合成并检测细胞凋亡率和凋亡蛋白Bax、Bcl-2表达的变化,明确神经酰胺作用机制。结果与野生型小鼠相比,WD小鼠肝内神经酰胺相关基因Smpd3和Cerk表达显著升高,差异均有统计学意义(P<0.05)。肝癌细胞内铜累积诱导细胞凋亡率显著升高,神经酰胺合成量也显著增加,差异均有统计学意义(P<0.05)。抑制神经酰胺的合成可通过调控凋亡蛋白Bax、Bcl-2的表达显著减少细胞凋亡率,差异有统计学意义(P<0.05)。结论WD中肝脏内累积的高铜可通过促进神经酰胺的合成调控下游凋亡效应分子Bax、Bcl-2的表达而引起肝细胞凋亡和肝脏损伤。Objective To investigate the regulatory effect of ceramide on Wilson’s disease(WD)copper accumulation induced hepatocyte apoptosis and its mechanism.Methods RNA-sequencing of the WD mice liver tissues was conducted,and the ceramide-related differentially expressed genes were enriched and verified by quantitative reverse transcription polymerase chain reaction(q RT-PCR).Gene MANIA database was used to construct the protein-protein interaction(PPI)between ceramide associated differential expressed proteins and apoptosis-related proteins.Hepatocellular carcinoma cells were treated with copper solutions at different concentrations,and the correlation of intracellular copper accumulation,cell apoptosis rate and intracellular ceramide content was analyzed.Myriocin was used to inhibit ceramide synthesis,and the changes of apoptosis rate and expression of apoptotic proteins Bax and Bcl-2 were detected to clarify the mechanism of ceramide action.Results Compared with wild-type mice,the expression of ceramide-related genes Smpd3 and Cerk in the liver of WD mice were significantly increased,the differences were statistically significant(P<0.05).Copper accumulation in hepatocellular carcinoma cells significantly increased apoptosis rate and ceramide synthesis,the differences were statistically significant(P<0.05).Inhibition of ceramide synthesis could significantly reduce the apoptosis rate by regulating the expression of apoptotic proteins Bax and Bcl-2,and the differences were statistically significant(P<0.05).Conclusion The copper accumulation in the liver of WD can induce hepatocyte apoptosis and liver injury by promoting ceramide synthesis and regulating the expression of downstream apoptotic proteins Bax and Bcl-2.

关 键 词：神经酰胺 威尔逊病 肝豆状核变性 铜累积 凋亡 

分 类 号：R742.4[医药卫生—神经病学与精神病学]