Clin Exp Otorhinolaryngol.  2016 Sep;9(3):244-251. 10.21053/ceo.2015.01011.

Radioprotective Effect of Epigallocatechin-3-Gallate on Salivary Gland Dysfunction After Radioiodine Ablation in a Murine Model

• Choi JS
• An HY
• Park IS
• Kim SK
• Kim YM
• Lim JY

• KMID: 2353623
• DOI: http://doi.org/10.21053/ceo.2015.01011

Abstract

OBJECTIVES
Radioiodine (RI) therapy is known to subject cellular components of salivary glands (SG) to oxidative stress leading to SG dysfunction. However, the protective effects of antioxidants on RI-induced SG damage have not been well investigated. The authors investigated the morphometric and functional effects of epigallocatechin-3-gallate (EGCG) administered prior to RI therapy and compared this with the effects of amifostine (a well-known antioxidant) in a murine model of RI sialadenitis.
METHODS
Four-week-old female C57BL/6 mice (n=48) were divided into four groups; a normal control group, a RI-treated group (0.01 mCi/g mouse, orally), an EGCG and RI-treated group, and an amifostine and RI-treated group. Animals in these groups were divided into 3 subgroups and euthanized at 15, 30, and 90 days post-RI treatment. Salivary flow rates and lag times were measured, and morphologic and histologic examinations and TUNEL (terminal deoxynucleotidyl transferase biotin-dUDP nick end labeling) assays were performed. Changes in salivary (99m)Tc pertechnetate uptake and excretion were followed by single-photon emission computed tomography.
RESULTS
Salivary flow rates and lag times to salivation in the EGCG or amifostine groups were better than in the RI-treated group. Histologic examinations of SGs in the EGCG or amifostine group showed more mucin-rich parenchyma and less periductal fibrosis than in the RI-treated group. Fewer apoptotic cells were observed in acini, ducts, and among endothelial cells in the EGCG or amifostine group than in the RI group. In addition, patterns of (99m)Tc pertechnetate excretion were quite different in the EGCG or amifostine group than in the RI group.
CONCLUSION
EGCG supplementation before RI therapy could protect from RI-induced SG damage in a manner comparable to amifostine, and thus, offers a possible means of preventing SG damage by RI.

Keyword

Radiation; Salivary Glands; Thyroid Neoplasms; Tomography, Emission-Computed, Single-Photon; Models, Animal

MeSH Terms

Amifostine
Animals
Antioxidants
DNA Nucleotidylexotransferase
Endothelial Cells
Female
Fibrosis
Humans
In Situ Nick-End Labeling
Mice
Models, Animal
Oxidative Stress
Salivary Glands*
Salivation
Sialadenitis
Sodium Pertechnetate Tc 99m
Thyroid Neoplasms
Tomography, Emission-Computed
Tomography, Emission-Computed, Single-Photon
Amifostine
Antioxidants
DNA Nucleotidylexotransferase
Sodium Pertechnetate Tc 99m

Figure

• 

  Fig. 1. Comparisons of mouse weights (A), salivary gland weights (B), salivary lag times (C), and flow rates (D). (A) Mice weight in the EGCG or amifostine treated groups was heavier. (B) SG weight in the normal control group was heavier. (C) Lag times and salivary flow rates in the EGCG or amifostine group were shorter and greater respectively. Kruskal-Wallis test and Dunn post hoc multiple comparison test (All P<0.05, respectively). Group I, the normal control; group II, RI exposed group; group III, administration of EGCG before RI exposure; group IV, administration of amifostine before RI exposure; EGCG, epigallocatechin-3-gallate; RI, radioiodine. a)Compared to group I. b)Compared to group II. C)Compared to group III.

• 

  Fig. 2. Histological analysis of salivary glands. Mucin-containing acini stained with Alcian blue appeared to be more numerous in the EGCG or amifostine group than in the RI group. MT staining showed the EGCG or amifostine group exhibited less periductal and perivascular fibrosis than the RI group. Group I, the normal control; group II, RI exposed group; group III, administration of EGCG before RI exposure; group IV, administration of amifostine before RI exposure; EGCG, epigallocatechin-3-gallate; RI, radioiodine; MT, Masson’s trichrome (scale bar, 20 μm).

• 

  Fig. 3. Histological analysis of tongues. Tongue staining showed irregular epithelium, a cornified layer of filiform papillae, and debris on tongue surfaces in the RI group (arrowhead). However, animals in the EGCG or amifostine group showed uniform, smooth epithelium, and clean tongue surfaces as compared with the RI group (arrow). (A) Group I, the normal control; (B) group II, RI exposed group; (C) group III, administration of EGCG before RI exposure; and (D) group IV, administration of amifostine before RI exposure. EGCG, epigallocatechin-3-gallate; RI, radioiodine (scale bar, 100 μm).

• 

  Fig. 4. Quantitative analysis by terminal deoxynucleotidyl transferase biotin-dUDP nick end labeling (TUNEL) assay. (A) Total numbers of TUNEL-positive cells, (B) TUNEL-positive acinar, (C) ductal cells, and (D) endothelial cells. (A) Analysis showed that total numbers of TUNEL-positive cells were significantly reduced in the EGCG or amifostine group as compared with RI group. (B–D) Numbers of TUNEL-positive acinar, ductal, and endothelial cells were significantly higher in the RI group and decreased in EGCG or amifostine treated group. Kruskal-Wallis test and Dunn’s post hoc multiple comparison test (all P<0.05, respectively). Group I, the normal control; group II, RI exposed group; group III, administration of EGCG before RI exposure; group IV, administration of amifostine before RI exposure; EGCG, epigallocatechin-3-gallate; RI, radioiodine. a)Compared to group I. b)Compared to group II. c)Compared to group III.

• 

  Fig. 5. Dynamics of 99mTc pertechnetate at 90 days post-RI. At 90 days posttreatment, 99mTc pertechnetate excretion was markedly lower in the RI-exposed group than other groups, but 99mexcretions in the EGCG or amifostine group were similar to that observed in the normal control group. Group I, the normal control; group II, RI exposed group; group III, administration of EGCG before RI exposure; group IV, administration of amifostine before RI exposure; EGCG, epigallocatechin-3-gallate; RI, radioiodine (Asterisks denote significant points).

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