Difference in H2O2 toxicity between intact renal tubules and cultured proximal tubular cells.
The present study was undertaken to examine the response to H2O2 and t-butylhydroperoxide (t-BHP) in various in vitro model systems of renal proximal tubules: rabbit renal cortical slices, freshly isolated rabbit proximal tubules, rabbit primary cultured proximal tubular cells, and opossum kidney (OK) cells. t-BHP increased lactate dehydrogenase release and lipid peroxidation in a concentration-dependent manner over the concentration range of 0.2 to 3 mM in cortical slices, whereas H2O2 caused a similar concentration-dependent increase in both parameters at 5-100 mM. The sensitivity of isolated tubules to both peroxides was similar to that of cortical slices. In primary cultured cells and OK cells, however, the cytotoxicity of H2O2 was identical to that of t-BHP. The cytotoxicity of t-BHP was not different among all the systems examined. The specific activity of catalase in cortical slices was similar to that of isolated tubules, but it was much higher than that of primary cultured cells or opossum kidney cells. Glutathione (GSH) peroxidase activity was not different among all the systems examined. The expression of catalase mRNA in cortical slices and isolated tubules was higher than that in primary cultured cells, whereas those of superoxide dismutase, glutathione peroxidase, or beta-actin were not different among the systems. These results indicate that intact proximal tubules are more resistant to H2O2 than are cultured proximal tubular cells, and the resistance is due to a higher specific activity of catalase resulting from the increased expression of its mRNA.
Kim
Y K
YK
Department of Physiology, College of Medicine, Pusan National University, Korea.
Ko
S H
SH
Woo
J S
JS
Lee
S H
SH
Jung
J S
JS
D000818:Animals; D002374:Catalase / Q000235:genetics; D002478:Cells, Cultured; D005979:Glutathione Peroxidase / Q000235:genetics; D006861:Hydrogen Peroxide / Q000633:toxicity*; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D010545:Peroxides / Q000633:toxicity; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D020122:tert-Butylhydroperoxide
D010545:Peroxides; D012333:RNA, Messenger; D020122:tert-Butylhydroperoxide; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D002374:Catalase; D005979:Glutathione Peroxidase