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. 2021 Jul;239(1):136-150.
doi: 10.1111/joa.13408. Epub 2021 Mar 13.

Cimetidine-induced androgenic failure causes cell death and changes in actin, EGF and V-ATPase immunoexpression in rat submandibular glands

Affiliations

Cimetidine-induced androgenic failure causes cell death and changes in actin, EGF and V-ATPase immunoexpression in rat submandibular glands

Mariane Castro Manzato et al. J Anat. 2021 Jul.

Abstract

Submandibular gland (SMG) is responsive to androgens via androgen receptor (AR). We verified whether cimetidine induces androgenic dysfunction in SMG, and evaluated the structural integrity, cell death and immunoexpression of actin, EGF and V-ATPase in androgen-deficient SMG. Male rats received cimetidine (CMTG) and control animals (CG) received saline. Granular convoluted tubules (GCTs) diameter and number of acinar cell nuclei were evaluated. TUNEL and immunofluorescence reactions for detection of AR, testosterone, actin, EGF and V-ATPase were quantitatively analysed. In CG, testosterone immunolabelling was detected in acinar and ductal cells cytoplasm. AR-immunolabelled nuclei were observed in acinar cells whereas ductal cells showed AR-immunostained cytoplasm, indicating a non-genomic AR action. In CMTG, the weak testosterone and AR immunoexpression confirmed cimetidine-induced androgenic failure. A high cell death index was correlated with decreased number of acinar cells, GCTs diameter and EGF immunoexpression under androgenic dysfunction. Actin immunofluorescence decreased in the SMG cells, but an increased and diffuse cytoplasmic V-ATPase immunolabelling was observed in striated ducts, suggesting a disruption in the actin-dependent V-ATPase recycling due to androgenic failure. Our findings reinforce the androgenic role in the maintenance of SMG histophysiology, and point to a potential clinical use of cimetidine against androgen-dependent glandular tumour cells.

Keywords: AR; antiandrogen; apoptosis; salivary glands; testosterone.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

FIGURE 1
FIGURE 1
(a–f) Photomicrographs of SMG sections of animals from CG and CMTG stained by H.E. In a and b, the glandular histoarchitecture shows numerous acini (A) and GCTs. In b, GCTs (double‐head arrows) are apparently reduced in comparison with CG. Either in b, d and f (CMTG), note reduced nuclear density in the acini (A), when compared with CG (a, c and e). In CMTG (d, f), either acinar cell nuclei (arrows), or GCT cell nuclei (arrowheads) are picnotic and/or irregularly outlined, in comparison with CG (c and e). Secretory granules (asterisks). Bars: 30 μm (a and b), 6 μm (c–f). (g) GCTs diameter (μm) in SMG of animals from CG and CMTG. (h) Numerical density of acinar cell nuclei/μm2 of glandular acini in CG and CMTG
FIGURE 2
FIGURE 2
(a–d) Photomicrographs of SMG sections subjected to TUNEL method and counterstained with haematoxylin. In a (CG), scarce TUNEL‐positive cells (arrows) are found in contrast with numerous TUNEL‐labelled cells (arrows) in b (CMTG). In c and d, TUNEL‐labelled cells are observed either in acini (thin arrows) or in GCTs (thick arrows). Bars: 54 μm (a and b), 6 μm (c and d), 16 μm (inset). (e) Number of TUNEL‐positive cells in SMG of rats from CG and CMTG
FIGURE 3
FIGURE 3
(a–f) Photomicrographs of SMG sections subjected to immunofluorescence reaction for detection of testosterone (c–f) and nuclear staining with DAPI (a and b). In c and e (CG), a cytoplasmic immunofluorescence is seen diffusely throughout the acinar cells (asterisks) whereas an evident testosterone immunoexpression is observed in the basal portion of GCT (arrows) and striated ductal (arrowheads) cells. In CMTG (d and f), no immunofluorescence is detected in the acini; the striated ducts (arrowheads) and GCTs (arrows) show scarce and weak signals of cytoplasmic immunolabelling. Bars: 10 μm. (g) Testosterone immunofluorescent area/μm2 of striated ducts in SMG of animals from CG and CMTG. (h) Testosterone immunofluorescent area/μm2 of GCT in SMG of animals from CG and CMTG
FIGURE 4
FIGURE 4
(a–f) Photomicrographs of SMG sections of animals from CG and CMTG subjected to immunofluorescence for detection of AR (c–f) and nuclear staining with DAPI (a and b). In c and e (CG), an intense immunofluorescence is noted in numerous acinar cell nuclei (arrows) whereas in d and f (CMTG), scarce nuclei are immunolabelled (arrows). Note that in CG (c and e), AR immunolabelling is also observed in the cytoplasm of GCT (thin arrows) and striated duct (arrowheads) cells. Bars: 14 μm. (g) AR immunofluorescent nuclear area/μm2 of acinar area in animals from CG and CMTG
FIGURE 5
FIGURE 5
(a–h) Photomicrographs of SMG sections subjected to immunofluorescence reaction for detection of actin (c–h) and nuclear staining with DAPI (a and b). In CG (c and e), a strong immunoreaction is observed throughout the structures (acini and ducts) of the glandular section, in comparison with the weak immunoexpression observed in CMTG (d and f). In g, note a strong and continuous immunofluorescence in the acinar periphery (thin arrows) and in the basal portion of GCTs (thick arrows) and striated ducts (arrowheads). The apical portion of striated duct cells is also immunolabelled (asterisks). On the other hand, in CMTG (h), a weak immunofluorescence is noted in some portions of the acinar periphery (thin arrows), and both GCTs (thick arrows) and striated ducts (arrowheads) also show weak immunoreaction. Bars: 30 μm (a–f), 14 μm (g and h). (i) Actin immunofluorescent area/μm2 of glandular tissue in the animals from CG and CMTG
FIGURE 6
FIGURE 6
(a–f) Photomicrographs of SMG sections subjected to EGF immunofluorescence reaction (c‐f) and nuclear staining with DAPI (a and b). In c–f, the EGF immunofluorescence is observed in the GCTs either in CG or CMTG. In c and e, intense EGF immunoreaction is observed filling the cytoplasm of all GCT cells (arrows) whereas in CMTG (d and f), a sparse immunofluorescence is noted only in some cells of the GCTs (arrows). Bars: 14 μm. (g) EGF immunofluorescent area/μm2 of GCT in SMG sections of animals from CG and CMTG
FIGURE 7
FIGURE 7
(a–f) Photomicrographs of SMG sections subjected to V‐ATPase immunofluorescence reaction (c‐f) and nuclear staining with DAPI (a and b). In c–f, V‐ATPase immunofluorescence is observed in the striated ducts (arrowheads) either in CG or CMTG. In CG (c and e), V‐ATPase immunoreaction is normally concentrated in the apical portion of striated duct cells (asterisks). In CMTG (d and f), a diffuse immunoreaction is observed throughout the cytoplasm of these cells (asterisks). Bars: 18 μm. (g) V‐ATPase immunofluorescent area/μm2 of striated duct in SMG sections of animals from CG and CMTG

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References

    1. Adişen, E. , Aral, A. , Aybay, C. & Gürer, M.A. (2008) Salivary epidermal growth factor levels in Behçet's disease and recurrent aphthous stomatitis. Dermatology, 217, 235–240. - PubMed
    1. Adthapanyawanich, K. , Kumchantuek, T. , Nakata, H. , Yamamoto, M. , Wakayama, T. , Nishiuchi, T. et al. (2015) Morphology and gene expression profile of the submandibular gland of androgen‐receptor‐deficient mice. Archives of Oral Biology, 60, 320–332. - PubMed
    1. Ahlner, B.H. , Hagelqvist, E. & Lind, M.G. (1994) Influence on rabbit submandibular gland injury by stimulation or inhibition of gland function during irradiation: Histology and morphometry after 15 gray. Annals of Otology, Rhinology & Laryngology, 103, 125–134. - PubMed
    1. Al‐Bataineh, M.M. , Gong, F. , Marciszyn, A.L. , Myerburg, M.M. & Pastor‐Soler, N.M. (2014) Regulation of proximal tubule vacuolar H+‐ATPase by PKA and AMP‐activated protein kinase. American Journal of Physiology ‐ Renal Physiology, 306, F981–F995. - PMC - PubMed
    1. Aldinucci, A. , Bonechi, E. , Manuelli, C. , Nosi, D. , Masini, E. , Passani, M.B. et al. (2016) Histamine regulates actin cytoskeleton in human toll‐like receptor 4‐activated monocyte‐derived dendritic cells tuning CD4+ T lymphocyte response. Journal of Biological Chemistry, 291, 14803–14814. - PMC - PubMed

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