镧离子和酸根离子在镧盐诱导HepG2细胞特定基因表达改变中的作用  

Roles of lanthanum ions and acid radical ions in changes of lanthanum salts-induced specific gene expressions in HepG2 cells

作　　者：邢云昆 陈洁 周小琰 马雪 周川 付娟玲[1] 祁妍敏[3] 郝卫东[1] 姚碧云[1] 赵鹏[1] XING Yun-kun;CHEN Jie;ZHOU Xiao-yan;MA Xue;ZHOU Chuan;FU Juan-ling;QI Yan-min;HAO Wei-dong;YAO Bi-yun;ZHAO Peng(Department of Toxicology,Beijing Key Laboratory of Toxicology Research and Risk Assessment for Food Safety,School of Public Health,Peking University,Beijing 100191,China;The Affiliated High School of Peking University,Beijing 100190,China;Civil Aviation Medicine Center,Civil Aviation Administration of China,Beijing 100123,China)

机构地区：[1]北京大学公共卫生学院毒理学系,食品安全毒理学研究与评价北京市重点实验室,北京100191 [2]北京大学附属中学,北京100190 [3]中国民用航空局民用航空医学中心,北京100123

出　　处：《中国药理学与毒理学杂志》2022年第12期910-918,共9页Chinese Journal of Pharmacology and Toxicology

基　　金：国家重点研发计划(2017YFC1600203);国家自然科学基金(81370079);国家自然科学基金(81001253);北京市自然科学基金(7132122)。

摘　　要：目的探究镧离子(La^(3+))与酸根离子(SO_(4)^(2-),Cl^(-)和NO_(3)^(-))在镧盐诱导Hep G2细胞特定基因表达改变中的作用。方法分别将含有La_(2)(SO_(4))_(3)0.71 mmol·L^(-1)、La Cl_(3)1.41 mmol·L^(-1)、La(NO_(3))_(3)1.41 mmol·L^(-1)、H_(2)SO_(4)2.12 mmol·L^(-1)、HCl 4.23 mmol·L^(-1)和HNO_(3)4.23 mmol·L^(-1)的培养液在CO_(2)培养箱中进行酸碱平衡1 h,p H值可恢复到7.25,随后分别处理对数生长期Hep G2细胞48 h。采用CCK-8试剂盒检测受试物对细胞存活率的影响。采用实时荧光定量PCR(RT-q PCR)检测丙氨酰氨基肽酶(ANPEP)、细胞色素P450家族1A1(CYP1A1)、氧化固醇结合蛋白样7(OSBPL7)、CYP3A5、钙电压门控通道辅助亚单位γ4(CACNG4)、双特异性磷酸酶4(DUSP4)、Ras蛋白特异性鸟嘌呤核苷酸释放因子1(RASGRF1)和CYP17A1 m RNA表达水平。结果CCK-8法检测结果显示,与细胞对照组比较,3种镧盐和HCl处理组细胞存活率均降低(P<0.01),而H_(2)SO_(4)和HNO_(3)处理组细胞存活率均未见明显改变。RT-q PCR结果显示,与细胞对照组比较,La_(2)(SO_(4))_(3),LaCl_(3)和La(NO_(3))_(3)均可诱导Hep G2细胞ANPEP,CYP1A1,OSBPL7,CYP3A5,CACNG4,DUSP4,RASGRF1和CYP17A1等8个基因m RNA表达上调(P<0.05);SO_(4)^(2-)诱导ANPEP和CYP1A1表达(P<0.05);Cl^(-)诱导CYP1A1表达(P<0.05);NO_(3)^(-)诱导ANPEP和CYP3A5 m RNA表达,而抑制CYP1A1和CACNG4 m RNA表达(P<0.05)。归因分析发现,La_(2)(SO_(4))_(3)和LaCl_(3)的La^(3+)均能诱导Hep G2细胞上述8个基因m RNA表达上调(P<0.05),但对CYP1A1,CYP3A5,DUSP4和CYP17A1m RNA表达的诱导作用低于La_(2)(SO_(4))_(3)(P<0.05),对CYP3A5和RASGRF1 m RNA表达的诱导作用低于LaCl_(3)(P<0.05);La(NO_(3))_(3)的La^(3+)虽能诱导上述8个基因m RNA表达上调,但ANPEP m RNA的表达上调无统计学意义,且对CYP3A5和CYP17A1 m RNA表达的诱导作用低于La(NO_(3))_(3)(P<0.05)。此外,3种镧盐的La^(3+)对ANPEP,OSBPL7,CYP3A5,CACNG4,DUSP4,RASGRF1和CYP17A1等7个基因m RNA表达的诱导作用彼此�OBJECTIVE To explore the roles of lanthanum ions(La^(3+))and acid radical ions(SO_(4)^(2-),Cl^(-)and NO_(3)^(-))in the expression changes of specific genes in Hep G2 cells induced by lanthanum salts,respectively.METHODS A culture medium containing 0.71 mmol·L^(-1)lanthanum sulfate﹝La_(2)(SO_(4))_(3)﹞,1.41 mmol·L^(-1)lanthanum chloride(LaCl_(3)),1.41 mmol·L^(-1)lanthanum nitrate﹝La(NO_(3))_(3)﹞,2.12 mmol·L^(-1)H_(2)SO_(4),4.23 mmol·L^(-1)HCl or 4.23 mmol·L^(-1)HNO_(3)was placed in a CO2incubator for acid-base balance for 1 h,and the p H value was restored to 7.25.Hep G2 cells in the logarithmic growth phase were treated with a acid-base-balanced culture medium containing the test chemical for 48 h.The effect of test chemicals on cell proliferation was detected by Cell Counting Kit-8(CCK-8)assay.The m RNA expression levels of the target genes[alanyl aminopeptidase(ANPEP),cytochrome P450 family 1 subfamily A member 1(CYP1A1),oxysterol binding protein like 7(OSBPL7),cytochrome P450 family 3 subfamily A member 5(CYP3A5),calcium voltage-gated channel auxiliary subunit gamma 4(CACNG4),dual specificity phosphatase 4(DUSP4),Ras protein specific guanine nucleotide releasing factor 1(RASGRF1)and cytochrome P450 family 17 subfamily A member 1(CYP17A1)]were determined by real-time quantitative PCR(RT-q PCR).RESULTS CCK-8 assay results showed that compared with the cell control group,the relative proliferation rate(RPR)of cells decreased in lanthanum salt-treated and HCl^(-)treated groups(P<0.01),but remained unchanged in the H_(2)SO_(4)-treated or HNO_(3)^(-)treated groups.RT-q PCR results showed that compared with the cell control group,La_(2)(SO_(4))_(3),LaCl_(3)and La(NO_(3))_(3)all induced up-regulation of 8 genes detected in Hep G2 cells(P<0.05),which were ANPEP,CYP1A1,OSBPL7,CYP3A5,CACNG4,DUSP4,RASGRF1 and CYP17A1,respectively.SO_(4)^(2-)induced up-regulation of ANPEP and CYP1A1(P<0.05)while Cl^(-)induced up-regulation of CYP1A1(P<0.05).NO_(3)^(-)induced the expressions of ANPEP and CYP3A5,but repressed th

关 键 词：硫酸镧 氯化镧 硝酸镧 酸根离子 细胞色素P450家族1A1 细胞 HEPG2 

分 类 号：R995[医药卫生—毒理学]