Toxic effects of non-steroidal anti-inflammatory drugs in a human intestinal epithelial cell line (HCT-8), as assessed by the MTT and neutral red assays

Allen, C.N.; Harpur, Ernest S.; Gray, T.J.B.; Hirst, Barry H.

doi:10.1016/0887-2333(91)90016-7

Cited by 30 publications

(9 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Compounds 1–3 were assayed at concentrations of 5 μg/mL, 10 μg/mL, 20 μg/mL, and 40 μg/mL against a human colorectal (HRT-18) cell line to determine their effects to mammalian cells in vitro, as described elsewhere 45 . Cells were cultured in RPMI-1640 medium with 10% fetal horse serum at 37 °C with 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Phenylthiazole Antibacterial Agents Targeting Cell Wall Synthesis Exhibit Potent Activity in Vitro and in Vivo against Vancomycin-Resistant Enterococci

Mohammad

Younis

et al. 2017

J. Med. Chem.

View full text Add to dashboard Cite

The emergence of antibiotic-resistant bacterial species, such as vancomycin-resistant enterococci (VRE), necessitates the development of new antimicrobials. Here, we investigate the spectrum of antibacterial activity of three phenylthiazole-substituted aminoguanidines. These compounds possess potent activity against VRE, inhibiting growth of clinical isolates at concentrations as low as 0.5 μg/mL. The compounds exerted a rapid bactericidal effect, targeting cell wall synthesis. Transposon mutagenesis suggested three possible targets: YubA, YubB (undecaprenyl diphosphate phosphatase (UPPP)) and YubD. Both UPPP as well as undecaprenyl diphosphate synthase were inhibited by compound 1. YubA and YubD are annotated as transporters and may also be targets since 1 collapsed the proton motive force in membrane vesicles. Using Caenorhabditis elegans, we demonstrate that two compounds (1, 3, at 20 μg/mL) retain potent activity in vivo, significantly reducing the burden of VRE in infected worms. Taken altogether, the results indicate that compounds 1 and 3 warrant further investigation as novel antibacterial agents against drug-resistant enterococci.

show abstract

Section: Methodsmentioning

confidence: 99%

Phenylthiazole Antibacterial Agents Targeting Cell Wall Synthesis Exhibit Potent Activity in Vitro and in Vivo against Vancomycin-Resistant Enterococci

Mohammad

Younis

et al. 2017

J. Med. Chem.

View full text Add to dashboard Cite

show abstract

“…The plates were then incubated for a further 20 min at 37°C. The cell monolayers were fixed by the further addition of 100 glA of buffered formalin, pH 7.0, to each well for 1 h. After overnight drying in air, the formazan product was extracted by addition of 100 pl acidified isopropanol to each well (1 ml 1 M HCI acid per 100 ml isopropanol) before absorbance at 570 nm was measured with a Dynatech MR700 ELISA plate reader (Allen et al, 1991).…”

Section: Measurement Of Bidirectional Transepithelial [3h]-vinblastinmentioning

confidence: 99%

“…This solution was then discarded and the plates were allowed to dry in air before the dye was extracted with a solution containing 1% acetic acid, 50% ethanol (1001l per well) over a period of 45 min with shaking. Absorbance was measured at 540 nm with an ELISA plate reader (Allen et al, 1991).…”

Section: Measurement Of Bidirectional Transepithelial [3h]-vinblastinmentioning

confidence: 99%

Epithelial secretion of vinblastine by human intestinal adenocarcinoma cell (HCT-8 and T84) layers expressing P-glycoprotein

Hunter

Hirst²,

Simmons³

1991

Br J Cancer

View full text Add to dashboard Cite

Summary P-glycoprotein expression was demonstrated in two human intestinal adenocarcinoma cell-lines ileocaecal and T84, colonic) by immunoprecipitation of a 170-180 kDa protein with monoclonal antibody JSB-1. Both HCT-8 and T84 formed functional epithelial cell layers of high transepithelial electrical resistance (>700 Q cm2) when grown on permeable matrices. These epithelial layers demonstrated vectorial secretion (net vinblastine fluxes in the basal-to-apical direction of 0.135 and 0.452 pmol h-' cm-2 in HCT-8 and T84 cell layers, respectively, from bathing solutions containing 10 nM vinblastine). These vectorial vinblastine secretions were sensitive to inhibition by verapamil. Passive transepithelial vinblastine permeation was limited by the presence of intercellular (tight) junctions, as demonstrated by the high transepithelial electrical resistance, and verapamil increased this passive vinblastine permeation concomitant with a reduction in the electrical resistance. Cellular vinblastine loading was significantly greater from the basal side, and this was also susceptible to inhibition by basal verapamil. The demonstration of vectorial transport of vinblastine in human intestinal colonic adenocarcinoma cell layers is direct evidence in favour of the hypothesis that the function of mdrl in epithelia from the gastrointestinal tract is to promote detoxification by a process of epithelial secretion. This study also highlights that cellular vinblastine accumulation depends not only upon P-glycoprotein function, but also upon differential apparent membrane permeabilities and the presence of intercellular (tight) junctions that may restrict drug permeation and cellular accumulation to apical or basal membrane domains.

show abstract

“…These findings, coupled with the observation that indomethacin-induced permeability changes occur within minutes, suggest that at least some of the alterations in mucosal integrity caused by indomethacin can be attributed to its direct local toxicity. Indomethacin is known from in vitro studies to be one of the NSAIDs most toxic to cultured epithelial cells (Allen et al 1991). It has been also shown that indomethacin can change the cellular synthesis of adenosine triphosphate through the anaerobic glycolytic pathway, a process which is known to increase epithelial tight junction permeability (Duffey et al 1981;Kurtel et al 1992).…”

Section: Resultsmentioning

confidence: 99%

Role of Nitric Oxide in Indomethacin‐Induced Gastric Mucosal Dysfunction in the Rat

Gürbüz

Ali̇can

Yeğen

et al. 1999

Experimental Physiology

View full text Add to dashboard Cite

summaryThe present study was undertaken to explore the role of nitric oxide (NO) in the pathogenesis of experimental non-steroidal anti-inflammatory drug (NSAID)-induced gastropathy. We assessed the role of NO inhibition and donation in indomethacin-induced gastric mucosal dysfunction. The stomach was perfused with vehicle (control) for 20 min, followed by indomethacin (10 mg ml¢ in 1·25% sodium bicarbonate, pH 8·4) for 120 min. N G -nitro-¬_arginine methyl ester (¬_NAME, 5 and 10 mg kg¢, i.v. bolus), ¬_arginine, ª_arginine (100 mg kg¢ i.v. bolus, 10 mg kg¢ h¢, 2 h infusion) and the NO donor glyceryl trinitrate (GTN) were given at the same time (20, 40 and 80 ìg kg¢ min¢, 15 min infusion) as perfusion with indomethacin was started. Epithelial permeability was quantified by measuring blood-to-lumen clearance of 51 Cr-labelled EDTA. Indomethacin caused a 20-fold increase in 51 Cr-EDTA leakage compared with that of the control group. Treatment with ¬_NAME or ¬_arginine did not affect the indomethacin-induced alterations in mucosal permeability. Administration of GTN (20 ìg kg¢ min¢) significantly reduced the indomethacin-induced mucosal dysfunction. By contrast, higher doses of GTN (80 ìg kg¢ min¢) exacerbated epithelial dysfunction induced by indomethacin. Elevated levels of carbonyls and myeloperoxidase (MPO) observed after indomethacin administration were significantly reduced, to the control values, when GTN (20 ìg kg¢ min¢) was administered along with indomethacin.

show abstract

Toxic effects of non-steroidal anti-inflammatory drugs in a human intestinal epithelial cell line (HCT-8), as assessed by the MTT and neutral red assays

Cited by 30 publications

References 27 publications

Phenylthiazole Antibacterial Agents Targeting Cell Wall Synthesis Exhibit Potent Activity in Vitro and in Vivo against Vancomycin-Resistant Enterococci

Phenylthiazole Antibacterial Agents Targeting Cell Wall Synthesis Exhibit Potent Activity in Vitro and in Vivo against Vancomycin-Resistant Enterococci

Epithelial secretion of vinblastine by human intestinal adenocarcinoma cell (HCT-8 and T84) layers expressing P-glycoprotein

Role of Nitric Oxide in Indomethacin‐Induced Gastric Mucosal Dysfunction in the Rat

Contact Info

Product

Resources

About