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《Chinese Pharmacological Bulletin》 2008-09
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Effects of solanine on kinetic constants of NATase in HepG_2 cells

JI Yu-bin1,2,GAO Shi-yong1,2,JI Chen-feng1,2,ZOU Xiang1,2(1.Postdoctoral Programme,Institute of Materia Medica,Center of Research and Development on Life Sciences andEnviroment Sciences of Harbin University of Commerce;2.Engineering Research Center ofNatural Anticancer Drugs,Ministry of Education.Harbin 150076,China)  
Aim To explore the effects of solanine on Km and Vmax of NATase of HepG2.Methods MTT assay was adopted to determine the cytotoxicity to SGC-7901,HepG2,and Ls-174.Employing HPLC,using 2-AF as substrate,taking concentration of 2-AF as concentration of substrate,in intact HepG2 cells and their cytoplasm,taking the speed of 2-AF being acetylated to 2-AFF by NATase as the rate of NATase,using double reciprocal plot,taking 1/S(the reciprocal of concentration of 2-AF) and 1/V(reaction rate of NATase) as coordinates,regression equation was obtanined,and Km and Vmax were calculated.Results Solanine was cytotoxic to HepG2.Study on enzyme kinetics demonstrated,as for intact HepG2 cells,Km and Vmax of control group were(2.37×10-3±8.37×10-5) mmol·L-1,(9.16×10-4±7.54×10-5) nmol·(106 cells)-1 respectively,Km and Vmax of the solanine group were(2.22×10-3±9.05×10-5) mmol·L-1,(5.14×10-4±3.72×10-5)nmol·(106 cells)-1 respectively.As for the cytoplasm of HepG2 cells,Km and Vmax of control group were(8.95×10-3±2.61×10-4) mmol·L-1 and(2.55×10-6±1.92×10-8)μmol·min-1·g-1 protein,Km and Vmax of the solanine group were(9.48×10-3±3.63×10-4) mmol·L-1 and(2.43×10-6±1.32×10-8) μmol·min-1·mg-1 protein.Statistically,as for intact HepG2 cells and their cytoplasm,there was no difference between the Km of control group and that of solanine group,but there was remarkable difference between Vmax of control group and that of solanine group,P0.01 for intact cell and P0.05 for cytoplasm.Conclusion Solanine was a non-competitive inhibitor of NATs in HepG2.
【Fund】: 国家自然科学基金资助项目(No30400591);; 黑龙江省自然科学基金资助项目(NoD200505 D-2004-13);; 黑龙江省教育厅骨干教师创新能力资助计划(2006);; 哈尔滨市青年基金资助项目(No2004AFQXJ035)
【CateGory Index】: R285.5;R735.7
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