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
作者
邢云昆
陈洁
周小琰
马雪
周川
付娟玲
祁妍敏
郝卫东
姚碧云
赵鹏
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)
出处
《中国药理学与毒理学杂志》
CAS
北大核心
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)。
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
lanthanum sulfate
lanthanum chloride
lanthanum nitrate
acid radical ion
cytochrome P450 family 1 subfamily A member 1
cells
HepG2
