Regulation of thioltransferase expression in human lens epithelial cells.

摘要: PURPOSE. To study how the expression of thioltransferase (TTase), a critical thiol repair and dethiolating enzyme, is regulated in human lens epithelial cells under oxidative stress. Also to examine whether depleting primary cellular antioxidant glutathione (GSH) these has any influence on TTase same conditions. METHODS. Human (B3) were grown confluence (1.6 million) gradually weaned from serum medium before exposing 0.1 mM H 2 O for hours. Cells removed at time intervals 0, 5, 10, 15, 30, 60, 120 minutes protein measurements GSH activity reverse transcription-polymerase chain reaction (RT-PCR) or Northern hybridization analysis quantify mRNA. The effect depletion mRNA was examined by treating with buthionine S,R-sulfoximine (BSO); 1-chloro, 2,4-dinitrobenzene (CDNB); 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU). Lens cells, depleted treatment BCNU, subjected stress level. RESULTS. A transient increase detected after 5 treatment. upregulation reached maximum 80% above normal level 10 decreased as oxidant detoxified cells. Manipulation BSO, CDNB, BCNU resulted minimum change expression. It noteworthy that when stress, also found be strongly upregulated. CONCLUSIONS. These observations suggest could an adaptive response combat restore vital functions proteins enzymes. Such regulation independent concentration.