title: N-acetylcysteine renoprotection in methotrexate induced nephrotoxicity and its effects on B-cell lymphoma.
abstract: BACKGROUND
  Nephrotoxicity is one of the known side effects of methotrexate (MTX) therapy despite the use of conventional protective measures. Our objectives were to evaluate the effects of N-acetylcysteine (NAC) on MTX-induced toxicity in renal tubular cells and to evaluate whether adjunctive use of NAC interferes with MTX antitumor activity in the B-cell lymphoma.
  
  
  METHODS
  Kidney Epithelial (Madin-Darby canine kidney [MDCK]) cells were exposed to MTX (10 μM or 100 μM) alone and with NAC (0.2 mM or 0.4 mM). Reactive oxygen species (ROS) generation at 1, 2, 4, and 24 h was measured by flow cytometer. Quantification of total glutathione (GSH) was performed by using GSH assay kit. To measure the impact of NAC on the antitumor activity of MTX, B lymphoma cells were exposed to MTX alone and with NAC. A percentage of apoptosis was measured using fluorescein isothiocyanate in both cell lines. Quantitative data was presented as a means ± standard deviation, and P values were analyzed using the Student's t-test.
  
  
  RESULTS
  Apoptosis in MDCK cells were observed after 24 h of incubation with both 10 μM and 100 μM MTX. Maximum ROS generation was observed at 4 h and corresponded to GSH production. Treatment with 0.2 and 0.4 mM of NAC led to decrease percentages of apoptotic MDCK cells. NAC did not change either proliferation or apoptosis of B-cell lymphoma.
  
  
  CONCLUSIONS
  Using NAC for kidney protection may not interfere with the antitumor activity of MTX. Further in vivo studies are warranted to confirm noninterference between MTX and NAC and assess synergistic antitumor effects.
authors.affiliation: Texas Tech Health Sciences Center, Amarillo, Texas 97106, USA.
authors.forename: Ruchi
authors.initials: R
authors.lastname: Singh
authors.affiliation: Texas Tech Health Sciences Center, Amarillo, Texas 97106, USA.
authors.forename: Rikin
authors.initials: R
authors.lastname: Shah
authors.affiliation: Texas Tech Health Sciences Center, Amarillo, Texas 97106, USA.
authors.forename: Curtis
authors.initials: C
authors.lastname: Turner
authors.affiliation: Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: Osvaldo
authors.initials: O
authors.lastname: Regueira
authors.affiliation: Texas Tech Health Sciences Center, Amarillo, Texas 97106, USA.
authors.forename: Tetyana L
authors.initials: TL
authors.lastname: Vasylyeva
keywords: B-cell lymphoma; Madin-Darby canine kidney epithelial cells; N-acetylcysteine; methotrexate; nephrotoxicity
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