Sulforaphane and its glutathione conjugate but not sulforaphane nitrile induce UDP-glucuronosyl transferase (UGT1A1) and glutathione transferase (GSTA1) in cultured cells.
Glucoraphanin in Brassica vegetables breaks down to either sulforaphane or sulforaphane nitrile depending on the conditions, and sulforaphane can be further conjugated with glutathione. Using a high-throughput microtitre plate assay and TaqMan real time quantitative RT-PCR to measure mRNA, we show that sulforaphane and its glutathione conjugate, but not the nitrile, increased significantly (P < 0.05) both UGT1A1 and GSTA1 mRNA levels in HepG2 and HT29 cells. These changes were accompanied by an increase in UGT1A1 protein, as assessed by immunoblotting, and a 2-8-fold increase in bilirubin glucuronidation. When treated together, the nitrile derivative did not affect sulforaphane induction. The induction of UGT1A1 and GSTA1 mRNA by sulforaphane was time and concentration dependent. The results show a functional induction of glucuronidation by sulforaphane but not sulforaphane nitrile, and show that the pathway of metabolism of glucosinolates in Brassica vegetables is important in determining the resulting biological and anticarcinogenic activities.
Basten
Graham P
GP
Institute of Food Research, Norwich Research Park, Conley Lane, Norwich, NR4 7UA, UK.
Bao
Yongping
Y
Williamson
Gary
G
D001483:Base Sequence; D001663:Bilirubin / Q000378:metabolism; D017931:DNA Primers; D004790:Enzyme Induction; D020719:Glucuronides / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000096:biosynthesis* / Q000235:genetics; D005978:Glutathione / Q000737:chemistry*; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D017879:Isothiocyanates; D009570:Nitriles / Q000737:chemistry; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013454:Sulfoxides; D013861:Thiocyanates / Q000737:chemistry / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D000099148:UGT1A1 Enzyme
D001663:Bilirubin; D017931:DNA Primers; D020719:Glucuronides; D014453:Glucuronosyltransferase; D005978:Glutathione; D005982:Glutathione Transferase; D017879:Isothiocyanates; D009570:Nitriles; D012333:RNA, Messenger; D013454:Sulfoxides; D013861:Thiocyanates; D000099148:UGT1A1 Enzyme; C016766:sulforaphane