A fluorescence-based analysis of aristolochic acid-derived DNA adducts

Abstract

Aristolochic acids (AAs), major components of plant extracts from Aristolochia species form, after metabolic activation, pro-mutagenic DNA adducts in renal tissue. The DNA adducts can be used as biomarkers for studies of AA toxicity. Identification of these adducts is a complicated and time-consuming procedure. We present herein a fast, non-isotopic, fluorescence-based assay for the detection of AA-DNA adducts in multiple samples. This approach allows analysis of AA adducts in synthetic DNA with known nucleotide composition, analysis of DNA adducts formed from chemically diverse AAs in vitro. The method can be applied to compare AA-DNA adduct formation in cells and tissues.

Defining in vivo dose‐response curves for kidney DNA adduct formation of aristolochic acid I in rat, mouse and human by an in vitro and physiologically based kinetic modeling approach

“…At confluency of 80%‐90%, the cells were exposed for 24 hours to AAI at different concentrations ranging from 0.5 to 20 μ m (final concentration in the DMEM without serum) added from 200 times concentrated stock solutions in dimethyl sulfoxide. In line with previous studies (Romanov, Sidorenko, Rosenquist, Whyard, & Grollman, 2012; Sidorenko et al, 2014), exposure of cells to AAI was performed in serum‐free medium to prevent binding of AAI to serum proteins, which would decrease the free concentration of AAI to which the cells were exposed (Dickman, Sweet, Bonala, Ray, & Wu, 2011).…”

“…In vitro DNA adduct formation was determined using LLC‐PK1 cells, which are proximal tubular cells from pig kidney that have been frequently used to assess in vitro AAI‐induced toxicity (Balachandran, Wei, Lin, Khan, & Pasco, 2005; Hsin et al, 2006; Romanov et al, 2012). Although AAI‐DNA adduct formation in pig cells may differ from AAI‐DNA adduct formation in kidney cells of rat, mouse and human, we predicted the DNA adduct levels for these three using this cell line.…”

Nitroreduction: A Critical Metabolic Pathway for Drugs, Environmental Pollutants, and Explosives

“…However, noninvasive and less expensive methodology will be required for biomonitoring. In this regard, a fluorescence-based assay has been developed for adduct detection …”

“…In this regard, a fluorescence-based assay has been developed for adduct detection. 139 10. FUTURE DIRECTIONS 10.1.…”

Gold nanoparticle-enhanced luminol/ferricyanide chemiluminescence system for aristolochic acid-I detection in medicinal plants and slimming products

“…The described CZE method displayed an outstanding detection of AA-I in Chinese medicine with detection limits of 0.6 and 0.05 µg mL −1 using CZE-UV and CZE-MS detectors, respectively. Furthermore, the literature survey reported a spectroscopic technique based on fluorescence analysis for the detection of AA-I derived DNA adducts in renal tissues (13). Finally, an electrochemical detection method based on square wave voltammetry was introduced to detect AA-I in Chinese medicinal plants with a limit of detection of 1.6 × 10 −8 mol L −1 (14).…”