PMID- 25822105
OWN - NLM
STAT- MEDLINE
DCOM- 20160111
LR  - 20171210
IS  - 1868-8551 (Electronic)
IS  - 1868-596X (Linking)
VI  - 32
IP  - 3
DP  - 2015
TI  - Simple and rapid in vitro assay for detecting human thyroid peroxidase 
      disruption.
PG  - 191-200
LID - 10.14573/altex.1412201 [doi]
AB  - A simple and rapid luminometric assay for the detection of chemical inhibitors of 
      human thyroid peroxidase (hTPO) activity was developed and validated with 10 
      model compounds. hTPO was derived from the human thyroid follicular cell line 
      Nthy-ori 3-1 and its activity was quantified by measuring the oxidation of 
      luminol in the presence of hydrogen peroxide (H2O2), which results in the 
      emission of light at 428 nm. In this assay,hTPO activity was shown to be 
      inhibited by 5 known TPO inhibitors and not inhibited by 5 non-inhibitors. 
      Similar results were obtained with porcine TPO (pTPO).The inhibition of hTPO by 
      the model compounds was also tested with guaiacol and Ampliflu Red as alternative 
      indicator substrates. While all substrates allowed the detection of pTPO activity 
      and its inhibition, only the Ampliflu Red and luminol-based methods were 
      sensitive enough to allow the quantification of hTPO activity from Nthy-ori 3-1 
      cell lysates. Moreover, luminol gave results with a narrower 95% confidence 
      interval and therefore more reliable data.Whole extracts of fast-growing Nthy-ori 
      3-1 cells circumvent the need for animal-derived thyroid organs,thereby reducing 
      costs, eliminating potential contamination and providing the possibility to study 
      human instead of porcine TPO. Overall, the application of luminol and Nthy-ori 
      3-1 cell lysate for the detection of the disruption of hTPO activity was found to 
      represent a valuable in vitro alternative and a possible candidate for inclusion 
      within a high throughput integrated testing strategy for the detection of 
      compounds that potentially interfere with normal thyroid function in vivo.
FAU - Jomaa, Barae
AU  - Jomaa B
AD  - Division of Toxicology, Wageningen University, Wageningen, The Netherlands.
FAU - de Haan, Laura H J
AU  - de Haan LH
AD  - Division of Toxicology, Wageningen University, Wageningen, The Netherlands.
FAU - Peijnenburg, Ad A C M
AU  - Peijnenburg AA
AD  - RIKILT - Institute of Food Safety, Wageningen University and Research Centre, 
      Wageningen, The Netherlands.
FAU - Bovee, Toine F H
AU  - Bovee TF
AD  - RIKILT - Institute of Food Safety, Wageningen University and Research Centre, 
      Wageningen, The Netherlands.
FAU - Aarts, Jac M M J G
AU  - Aarts JM
AD  - Division of Toxicology, Wageningen University, Wageningen, The Netherlands.
FAU - Rietjens, Ivonne M C M
AU  - Rietjens IM
AD  - Division of Toxicology, Wageningen University, Wageningen, The Netherlands.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150330
PL  - Germany
TA  - ALTEX
JT  - ALTEX
JID - 100953980
RN  - 0 (Antithyroid Agents)
RN  - 0 (Autoantigens)
RN  - 0 (Iron-Binding Proteins)
RN  - 0 (Luminescent Agents)
RN  - 0 (Thyroid Hormones)
RN  - 5EXP385Q4F (Luminol)
RN  - EC 1.11.1.7 (TPO protein, human)
RN  - EC 1.11.1.8 (Iodide Peroxidase)
SB  - IM
MH  - *Animal Testing Alternatives
MH  - Animals
MH  - *Antithyroid Agents
MH  - Autoantigens/metabolism
MH  - Cells, Cultured
MH  - Humans
MH  - In Vitro Techniques
MH  - Iodide Peroxidase/*antagonists & inhibitors/metabolism
MH  - Iron-Binding Proteins/*antagonists & inhibitors/metabolism
MH  - Luminescent Agents/chemistry
MH  - Luminol/chemistry
MH  - Swine
MH  - Thyroid Gland/drug effects
MH  - Thyroid Hormones/metabolism
OTO - NOTNLM
OT  - Nthy-ori 3-1
OT  - alternatives to animal testing
OT  - human thyroid peroxidase
OT  - in vitro assay
OT  - thyroid disruption
EDAT- 2015/03/31 06:00
MHDA- 2016/01/12 06:00
CRDT- 2015/03/31 06:00
PHST- 2014/12/20 00:00 [received]
PHST- 2015/03/24 00:00 [accepted]
PHST- 2015/03/31 06:00 [entrez]
PHST- 2015/03/31 06:00 [pubmed]
PHST- 2016/01/12 06:00 [medline]
AID - 10.14573/altex.1412201 [doi]
PST - ppublish
SO  - ALTEX. 2015;32(3):191-200. doi: 10.14573/altex.1412201. Epub 2015 Mar 30.