镉暴露诱导小鼠脾脏免疫相关差异表达基因的分析与鉴定  

Analysis and identification of differentially expressed genes related to spleen immunity induced by cadmium exposure in mice

作　　者：阮嘉诚 史美琳 马海燕 施海峰[1] RUAN Jiacheng;SHI Meilin;MA Haiyan;SHI Haifeng(College of Life Sciences,Jiangsu University,Zhenjiang 212013,China)

机构地区：[1]江苏大学生命科学学院,镇江212013

出　　处：《华夏医学》2024年第1期9-17,共9页Acta Medicinae Sinica

基　　金：国家自然科学基金项目(31071030,31271272)。

摘　　要：目的本研究探讨镉暴露对脾组织的损伤和基因表达的影响,并探究镉免疫毒性的潜在分子机制。方法采用单次皮下注射氯化镉(5μmol/kg)建立镉暴露小鼠模型,使用Synergy H4混合多模酶标仪检测脾组织中丙二醛(MDA)含量和人B淋巴细胞瘤细胞中Ca^(2+)含量,采用DNA基因芯片技术检测小鼠脾脏组织中差异表达基因,采用GO富集、KEGG信号通路和Reactome数据库分析镉暴露干扰的免疫相关功能,采用RT-qPCR、免疫荧光和免疫印迹等方法验证重要的免疫相关差异表达基因。结果镉暴露干扰细胞的自噬功能,引起细胞Ca^(2+)稳态失衡和组织氧化应激增强。镉暴露产生免疫相关脾组织差异表达基因934个,其中上调的基因687个,下调的基因247个。镉暴露干扰调节先天性和适应性免疫系统的4个关键基因Ppp3ca、Plcg2、Vav1和Cd247表达。结论镉暴露引起免疫调节关键基因Ppp3ca、Plcg2、Vav1、Cd247的表达改变可能是小鼠脾脏产生免疫毒性的分子机制。Objective This study aims to explore the effects of cadmium(Cd)exposure on spleen tissue damage and gene expression,and explore the underlying molecular mechanisms of cadmium immunotoxicity.Methods A single subcutaneous injection of CdCl 2(5μmol/kg)was used to expose the mouse model.The Synergy H4 hybrid microplate reader was used to detect malondialdehyde(MDA)content in spleen tissue and Ca^(2+)content in human B lymphocytoma cells,and the differentially expressed genes in the spleen were identified by DNA microarray analysis.GO enrichment,KEGG sinaling,and Reactome pathway database were used to analyze the immune related functions of Cd exposure interference.RT-qPCR,immunofluorescence and Western blot were used to differentially expressed genes.Results Cd exposure interferes with the autophagic function of cells,causing Ca^(2+)homeostasis imbalance and enhanced oxidative stress in tissues.There were 934 differentially expressed immune-related genes,including 687 up-regulated genes and 247 down-regulated genes.Cd-induced four key immune genes(Ppp3ca,Plcg2,Vav1,Cd247)regulated the innate and adaptive immune systems.Conclusion The expression changes of key immune regulatory genes(Ppp3ca,Plcg2,Vav1,and Cd247)induced by cadmium exposure may be the molecular mechanism of their immunotoxicity to mouse spleen.

关 键 词：镉 DNA芯片 免疫毒性 差异表达基因 

分 类 号：R995[医药卫生—毒理学]