四种硝基多环芳烃对HepaRG肝细胞脂质代谢影响的研究  

Effects of four nitrated polycyclic aromatic hydrocarbons on lipid metabolism in HepaRG liver cells

作　　者：龙玥含 何颖阳 顾雯[1] 石莹[1] 詹星宇 王开宜 陈圆圆 段链[1] LONG Yue-han;HE Ying-yang;GU Wen;SHI Ying;ZHAN Xing-yu;WANG Kai-yi;CHEN Yuan-yuan;DUAN Lian(China CDC Key Laboratory of Environment and Population Health/National Institute of Environmental Health,Chinese Center for Disease Control and Prevention,Beijing 100021,China;Institute of Medical Information,Chinese Academy of Medical Sciences)

机构地区：[1]中国疾病预防控制中心环境与人群健康重点实验室/中国疾病预防控制中心环境与健康相关产品安全所,北京100021 [2]中国医学科学院医学信息研究所

出　　处：《环境卫生学杂志》2025年第4期286-293,共8页JOURNAL OF ENVIRONMENTAL HYGIENE

基　　金：国家自然科学基金青年基金(22206178)。

摘　　要：目的探讨1-硝基芘、3-硝基荧蒽、1-硝基萘和9-硝基蒽4种硝基多环芳烃(nitrated polycyclic aromatic hydrocarbons,NPAHs)对HepaRG肝细胞的毒性作用及其对脂质代谢的影响。方法利用GlutaMAXTM添加剂和HepaRGTM无血清诱导培养基补充剂诱导细胞分化3 d,通过实时荧光定量PCR(quantitative real-time polymerase chain reaction,qRT-PCR)方法检测细胞药物代谢酶基因表达水平(CYP2B6、CYP3A4、GSTA1/2和UGT1A1)和吸光度法检测细胞白蛋白(albumin,ALB)水平,验证HepaRG肝细胞分化模型构建情况。基于上述细胞分化模型,以不同剂量1-硝基芘、3-硝基荧蒽、1-硝基萘和9-硝基蒽作为受试物暴露染毒48 h,通过吸光度法检测染毒后分化HepaRG细胞存活率、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)、总胆固醇(total cholesterol,TC)和甘油三酯(triacylglycerol,TG)相对表达倍数;qRT-PCR方法检测脂质代谢相关基因FABP4、LPL和PPARG的相对表达水平。结果与未分化HepaRG细胞相比,诱导3 d后HepaRG细胞药物代谢相关基因CYP2B6、CYP3A4、GSTA1/2和UGT1A1表达水平显著上调,ALB水平明显上升,成功构建具有代谢功能的肝细胞模型。以含有0.1%DMSO暴露染毒的实验组为对照组,与对照组相比,4种NPAHs不同剂量范围暴露48 h后均导致细胞活性降低且具有剂量依赖性,AST、ALP、TC和TG相对表达倍数增加以及脂质代谢相关基因FABP4、LPL和PPARG显著上调。结论硝基多环芳烃能够导致肝细胞损伤,可能通过干扰脂质代谢稳态而引起脂质蓄积。Objective To investigate the toxic effects of four nitrated polycyclic aromatic hydrocarbons(NPAHs)including 1-nitropyrene,3-nitrofluoranthene,1-nitronaphthalene,and 9-nitroanthracene on HepaRG liver cells and their effects on lipid metabolism.Methods HepaRG liver cells were differentiated for 3 days using the GlutaMAXTM Supplement and HepaRGTM Serum-free Induction Medium Supplement.The expression levels of drug-metabolizing enzyme genes(CYP2B6,CYP3A4,GSTA1/2,and UGT1A1)were determined by quantitative real-time polymerase chain reaction(qRT-PCR),and the levels of albumin(ALB)were measured by absorbance to validate the construction of HepaRG liver cell differentiation model.Different doses of 1-nitropyrene,3-nitrofluoranthene,1-nitronaphthalene,and 9-nitroanthracene were used to treat the cell differentiation model,with exposure to these compounds for 48 hours.The survival rate of differentiated HepaRG cells after exposure to poisonous substances,along with the relative expression levels of aspartate aminotransferase(AST),alkaline phosphatase(ALP),total cholesterol(TC),and triacylglycerol(TG)was determined by absorbance.The relative expression levels of lipid metabolism-related genes FABP4,LPL,and PPARG were determined using the qRT-PCR method.Results Compared with undifferentiated HepaRG cells,the expression levels of drug metabolism-related genes CYP2B6,CYP3A4,GSTA1/2,and UGT1A1 were significantly upregulated in HepaRG cells,and the level of ALB was significantly increased after 3 days of induction.The above results indicated that a liver cell model with metabolic function was successfully constructed in this study.Using differentiated HepaRG exposed to 0.1%DMSO as the control,differentiated HepaRG exposed to the four NPAHs at different dose ranges for 48 hours resulted in a dose-dependent decrease in cell viability,an increase in the relative expres-sion levels of AST,ALP,TC,and TG,and a significant upregulation of lipid metabolism-related genes FABP4,LPL and PPARG.ConclusionNPAHs cause hepatocellular injury,wh

关 键 词：硝基多环芳烃 肝损伤 脂质代谢紊乱 

分 类 号：R12[医药卫生—环境卫生学] R114[医药卫生—公共卫生与预防医学]