Endocrinol Metab.  2016 Mar;31(1):174-184. 10.3803/EnM.2016.31.1.174.

Propylthiouracil, Perchlorate, and Thyroid-Stimulating Hormone Modulate High Concentrations of Iodide Instigated Mitochondrial Superoxide Production in the Thyroids of Metallothionein I/II Knockout Mice

• Duan Q
• Wang T
• Zhang N
• Perera V
• Liang X
• Abeysekera IR
• Yao X

• KMID: 2169710
• DOI: http://doi.org/10.3803/EnM.2016.31.1.174

Abstract

BACKGROUND
Increased oxidative stress has been suggested as one of the underlying mechanisms in iodide excess-induced thyroid disease. Metallothioneins (MTs) are regarded as scavengers of reactive oxygen species (ROS) in oxidative stress. Our aim is to investigate the effects of propylthiouracil (PTU), a thyroid peroxidase inhibitor, perchlorate (KClO4), a competitive inhibitor of iodide transport, and thyroid stimulating hormone (TSH) on mitochondrial superoxide production instigated by high concentrations of iodide in the thyroids of MT-I/II knockout (MT-I/II KO) mice.
METHODS
Eight-week-old 129S7/SvEvBrd-Mt1(tm1Bri) Mt2(tm1Bri)/J (MT-I/II KO) mice and background-matched wild type (WT) mice were used.
RESULTS
By using a mitochondrial superoxide indicator (MitoSOX Red), lactate dehydrogenase (LDH) release, and methyl thiazolyl tetrazolium (MTT) assay, we demonstrated that the decreased relative viability and increased LDH release and mitochondrial superoxide production induced by potassium iodide (100 µM) can be relieved by 300 µM PTU, 30 µM KClO4, or 10 U/L TSH in the thyroid cell suspensions of both MT-I/II KO and WT mice (P<0.05). Compared to the WT mice, a significant decrease in the relative viability along with a significant increase in LDH release and mitochondrial superoxide production were detected in MT-I/II KO mice(P<0.05).
CONCLUSION
We concluded that PTU, KClO4, or TSH relieved the mitochondrial oxidative stress induced by high concentrations of iodide in the thyroids of both MT-I/II KO and WT mice. MT-I/II showed antioxidant effects against high concentrations of iodide-induced mitochondrial superoxide production in the thyroid.

Keyword

Propylthiouracil; Perchlorate; Thyrotropin; Metallothionein I/II knockout; Iodides

MeSH Terms

Animals
Antioxidants
Iodide Peroxidase
Iodides
L-Lactate Dehydrogenase
Metallothionein*
Mice
Mice, Knockout*
Oxidative Stress
Potassium Iodide
Propylthiouracil*
Reactive Oxygen Species
Superoxides*
Suspensions
Thyroid Diseases
Thyroid Gland*
Thyrotropin*
Antioxidants
Iodide Peroxidase
Iodides
L-Lactate Dehydrogenase
Metallothionein
Potassium Iodide
Propylthiouracil
Reactive Oxygen Species
Superoxides
Suspensions
Thyrotropin

Figure

• 

  Fig. 1 Effect of propylthiouracil (PTU) on high concentrations of iodide induced relative viability, lactate dehydrogenase (LDH) release and mitochondrial superoxide production in metallothionein I/II knockout (MT-I/II KO) and wild type (WT) mice. (A, B) Effects of PTU on high concentrations of iodide induced mitochondrial superoxide production. Histogram analysis was performed on the mean fluorescence intensity of MitoSOX Red as measured by flow cytometry. Experiments were repeated 3 times with similar results. (C) Effects of PTU on high concentrations of iodide induced LDH release. (D) Effects of PTU on high concentrations of iodide induced relative viability. One-way analysis of variance with a least significant difference test was used. aP<0.05 compared with the control group of WT or MT-I/II KO mice respectively; bP<0.05 compared with the potassium iodide (KI) group of WT or MT-I/II KO mice respectively; cP<0.05, WT mice compared with the MT-I/II KO mice under the same treatment.

• 

  Fig. 2 Effect of perchlorate (KClO4) on high concentrations of iodide induced relative viability, lactate dehydrogenase (LDH) release and mitochondrial superoxide production in metallothionein I/II knockout (MT-I/II KO) and wild type (WT) mice. (A, B) Effects of KClO4 on high concentrations of iodide induced mitochondrial superoxide production. Histogram analysis was performed on the mean fluorescence intensity of MitoSOX Red as measured by flow cytometry. Experiments were repeated 3 times with similar results. (C) Effects of KClO4 on high concentrations of iodide induced LDH release. (D) Effects of KClO4 on high concentrations of iodide induced relative viability. One-way analysis of variance with a least significant difference test was used. aP<0.05 compared with the control group of WT or MT-I/II KO mice respectively; bP<0.05 compared with the potassium iodide (KI) group of WT or MT-I/II KO mice respectively; cP<0.05 WT mice compared with the MT-I/II KO mice under the same treatment.

• 

  Fig. 3 Effect of thyroid stimulating hormone (TSH) on high concentrations of iodide induced relative viability, lactate dehydrogenase (LDH) release and mitochondrial superoxide production in metallothionein I/II knockout (MT-I/II KO) and wild type (WT) mice. (A, B) Effects of TSH on high concentrations of iodide induced mitochondrial superoxide production. Histogram analysis was performed on the mean fluorescence intensity of MitoSOX Red as measured by flow cytometry. Experiments were repeated 3 times with similar results. (C) Effects of TSH on high concentrations of iodide induced LDH release. (D) Effects of TSH on high concentrations of iodide induced relative viability. One-way analysis of variance with a least significant difference test was used. aP<0.05 compared with the control group of WT or MT-I/II KO mice respectively; bP<0.05 compared with the potassium iodide (KI) group of WT or MT-I/II KO mice respectively; cP<0.05 WT mice compared with the MT-I/II KO mice under the same treatment.

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