Parola M, Biocca M E, Leonarduzzi G, Albano E, Dianzani M U, Gilmore K S, Meyer D J, Ketterer B, Slater T F, Cheeseman K H
Dipartimento di Medicina ed Oncologia Sperimentale, Università di Torino, Italy.
Biochem J. 1993 Apr 15;291 ( Pt 2)(Pt 2):641-7. doi: 10.1042/bj2910641.
The constitutive and inducible cytosolic glutathione S-transferase (EC 2.5.1.18) subunit compositions of parenchymal cells (hepatocytes) and biliary epithelial cells (BEC) from rat liver have been quantitatively analysed using reverse-phase h.p.l.c. Hepatocytes, analysed in the absence of non-parenchymal cells, expressed constitutively the following subunits, in order of their concentration: 3, 4, 2, 1a, 1b, 8, 6 and 10. BEC express constitutively only four of the GST subunits expressed by hepatocytes and these are, in order of their concentration: subunits 2, 7, 4 and 3. Notable differences from hepatocytes are that BEC completely lack the Alpha-class subunits 1a and 1b that are major subunits in hepatocytes, Mu-class subunits make up a very low proportion of the total, and the Pi-class subunit 7 is a major subunit in BEC, whereas it is essentially absent from hepatocytes. For the first time, the effects of the inducing agents phenobarbitone (PB), beta-naphthoflavone (beta-NF) and ethoxyquin (EQ) have been characterized in a comprehensive and quantitative manner in both cell types. PB, beta-NF and EQ increased total GST protein in hepatocytes by approx. 2-fold, 3-fold and 4-fold respectively. Subunits significantly induced in hepatocytes were (in order of fold-induction): by PB, 1b > 8 > 3 > 2 > 4; by beta-NF, 1b > 8 > 2 > 3 > 4; and by EQ, 7 > 1b > 10 > 8 > 3 > 2 > 1a > 4. In BEC, neither PB nor beta-NF had significant effects on the total amount of GST protein, although PB did significantly induce subunit 3 at the expense of other subunits. EQ increased total GST protein nearly 5-fold in BEC, subunits 7 and 3 being induced dramatically above constitutive levels.
采用反相高效液相色谱法对大鼠肝脏实质细胞(肝细胞)和胆管上皮细胞(BEC)中组成型和诱导型胞质谷胱甘肽S-转移酶(EC 2.5.1.18)的亚基组成进行了定量分析。在无非实质细胞的情况下分析肝细胞,其组成型表达以下亚基,按浓度顺序为:3、4、2、1a、1b、8、6和10。BEC仅组成型表达肝细胞所表达的GST亚基中的四个,按浓度顺序为:亚基2、7、4和3。与肝细胞的显著差异在于,BEC完全缺乏肝细胞中的主要亚基α类亚基1a和1b,μ类亚基在总量中所占比例极低,而π类亚基7是BEC中的主要亚基,在肝细胞中基本不存在。首次以全面和定量的方式对诱导剂苯巴比妥(PB)、β-萘黄酮(β-NF)和乙氧喹(EQ)在这两种细胞类型中的作用进行了表征。PB、β-NF和EQ使肝细胞中的总GST蛋白分别增加了约2倍、3倍和4倍。在肝细胞中显著诱导的亚基(按诱导倍数顺序)为:PB诱导的1b>8>3>2>4;β-NF诱导的1b>8>2>3>4;EQ诱导的7>1b>10>8>3>2>1a>4。在BEC中,PB和β-NF对GST蛋白总量均无显著影响,尽管PB确实以牺牲其他亚基为代价显著诱导了亚基3。EQ使BEC中的总GST蛋白增加了近5倍,亚基7和3被显著诱导至高于组成型水平。