Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Sep 3;16(9):e68515.
doi: 10.7759/cureus.68515. eCollection 2024 Sep.

Melatonin Mitigates Cisplatin-Induced Submandibular Gland Damage by Inhibiting Oxidative Stress, Inflammation, Apoptosis, and Fibrosis

Alaa M Badawy  1 , Mohie Ibrahim  2   1 , Medhat Taha  3   1 , Azza I Helal  4 , Ahmed Abdel-Monem Elmetwally  5 , Ibrahim El-Shenbaby  5 , Sara Abubakr  1 , Emadeldeen Hussin  1 , Noha Hammad Sakr  6 , Tourki A S Baokbah  7 , Amira E Farage  6
Affiliations

Melatonin Mitigates Cisplatin-Induced Submandibular Gland Damage by Inhibiting Oxidative Stress, Inflammation, Apoptosis, and Fibrosis

Alaa M Badawy et al. Cureus. .

Abstract

Background: The study aims to examine the possible effect of melatonin against cisplatin-induced submandibular degeneration in experimental rats exploring its ameliorative mechanisms.

Methods: Rats were classified into four experimental groups; control group; melatonin group; cisplatin group; and cisplatin+melatonin group. Submandibular tissues were collected. Biochemical, histopathological, and immunohistopathological examination and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis were performed.

Results: The results indicate that intraperitoneal administration of melatonin (30 mg/kg body weight) alongside cisplatin significantly elevated submandibular glands (SMG) and reduced glutathione (GSH) and superoxide dismutase (SOD) levels (p < 0.001), while it reduced malondialdehyde (MDA) levels, NF-κB gene expression, the protein level of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), immunoexpression of low-dose cyclooxygenase-2 (Cox-2), and CD68. Moreover, melatonin reduced immune and gene expression of alpha-smooth muscle actin (α-SMA), immunoexpression of caspase-3, and gene expression of Bax in comparison to the cisplatin group.

Conclusion: Melatonin attenuated cisplatin-induced submandibular destruction alleviating SMG oxidative stress, inflammation, and fibrosis in addition to halting cellular apoptosis, sheds light on its usage in clinical application.

Keywords: apoptosis; cisplatin; fibrosis; inflammation; melatonin; oxidative stress; submandibular gland.

PubMed Disclaimer

Conflict of interest statement

Human subjects: All authors have confirmed that this study did not involve human participants or tissue. Animal subjects: The study was conducted in accordance with the Canadian Council on Animal Care Guidelines and approved by the Committee of Research Ethics, Kafrelsheikh University Issued protocol number KFS-IACUC/204/2024. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work.

Figures

Figure 1
Figure 1. Representative photomicrograph of the submandibular gland from different treatment groups.
(A, B) Control and melatonin group showing the normal histological architecture of glandular acini. (C) Cisplatin group showing marked dilation of mucous acini with pressure on interstitial connective tissue (thick arrow). (D) Cisplatin showing marked replacement of acinar architecture with cellular mass formed of abundant lymphoplasmacytic cells (thin arrow). (E) Cisplatin+melatonin showing focal peri-glandular aggregation of inflammatory cells (thin arrow). Image magnification: 400x
Figure 2
Figure 2. Markers of oxidative stress in submandibular glands of different experimental groups ((A) MDA, (B) GSH, and (C) SOD).
Data were expressed in mean±SD. !!! indicates p < 0.001 significant versus control rats; ~~~ indicates p < 0.001 significant versus cisplatin group. MDA: malondialdehyde; GSH: glutathione; SOD: superoxide dismutase; SMG: submandibular gland
Figure 3
Figure 3. Effect of melatonin on cisplatin-induced inflammatory markers: (A) Gene expression of NF-κB. (B, C, D) Protein level of TNF-α, IL-6, IL-1β by Elisa assay.
Data were expressed in mean±SD. !!! indicates p < 0.001 significant versus control rats; ~~~ indicates p < 0.001 significant versus cisplatin group. NF-κB: nuclear factor kappa B; TNF-α: tumor necrosis factor-alpha; IL-6: interleukin-6; IL-1β: interleukin-1 beta; SMG: submandibular gland
Figure 4
Figure 4. Representative IHC of Cox-2 expression in submandibular gland sections of different treatment groups.
(A) Normal group showing few immunoreactivities for Cox-2 in peri-glandular cells. (B) Melatonin group showing few immunoreactivities for Cox-2 in interstitial cells. (C, D) Cisplatin group revealing moderate to high immunoreactivity for Cox in peri-glandular cells with cytoplasmic expression in invading cellular mass. (E) Cisplatin+melatonin showing minimal immunoreactivity for Cox-2 in peri-glandular cells. Image magnification: 400x; arrowhead: positive glandular epithelial cells; thick arrow: positive cellular mass; thin arrow: positive interstitial cells. (F) Quantitative analysis of Cox-2 immunostaining (% area). Data were expressed in mean±SD. !!! indicates p < 0.001 significant versus control rats; ~~~ indicates p < 0.001 significant versus cisplatin group. IHC: immunohistochemistry
Figure 5
Figure 5. Representative IHC of CD68 expression in submandibular gland sections of different treatment groups.
(A, B) Normal and melatonin groups showing few immunoreactivities for CD68 in peri-glandular cells. (C, D) Cisplatin group showing moderate to high immunoreactivity for CD68 in peri-glandular cells and invading cellular mass. (E) Cisplatin+melatonin showing minimal to mild immunoreactivity for CD68 in peri-glandular cells. Image magnification: 400x; thick arrow: positive cellular mass; thin arrow: positive interstitial cells. (F) Quantitative analysis of CD68 immunostaining (% area). Data were expressed in mean±SD. !!! indicates p < 0.001 significant versus control rats; ~~~ indicates p < 0.001 significant versus cisplatin group. IHC: immunohistochemistry
Figure 6
Figure 6. Representative IHC of α-SMA expression in submandibular gland sections of different treatment groups.
(A) Normal group showing mild to moderate positive expression of fibril α-SMA around the glandular acini. (B) Melatonin group showing mild to moderate immunostained myocytes. (C, D) Cisplatin group showing high positive nuclear and cytoplasmic staining of invading cells with fibril expression around the dilated gland. (E) Cisplatin+melatonin showing moderate immunostaining in peri-glandular cells. Image magnification: 400x; thin arrow: positive interstitial tissue; thick arrow: positive inflammatory cell. (F) Quantitative analysis of α-SMA immunostaining (% area). (G) Gene expression of α-SMA. Data were expressed in mean±SD. !!! indicates p < 0.001 significant versus control rats; ~~~ indicates p < 0.001 significant versus cisplatin group. IHC: immunohistochemistry; α-SMA: α-smooth muscle actin
Figure 7
Figure 7. Representative IHC of TGF-β expression in submandibular gland sections of different treatment groups.
(A, B) Normal and melatonin groups showing few nuclear immunoreactivities for TGF-β in glandular and peri-glandular cells. (C, D) Cisplatin group showing moderate nuclear with cytoplasmic immunoreactivity for TGF-β in glandular cells and invading cellular mass. (E) Cisplatin+melatonin showing minimal to mild nuclear immunoreactivity for TGF-β in peri-glandular cells. Image magnification: 400x; arrowhead: positive glandular epithelial cells; thick arrow: positive cellular mass; thin arrow: positive interstitial cells. (F) Quantitative analysis of TGF-β immunostaining (%area). (G) Gene expression of α TGF-β. Data were expressed in mean±SD. !!! indicates p < 0.001 significant versus control rats; ~~~ indicates p < 0.001 significant versus cisplatin group. IHC: immunohistochemistry; TGF-β: transforming growth factor beta
Figure 8
Figure 8. Representative IHC of caspase-3 expression in submandibular gland sections of different treatment groups.
(A) Normal group showing few immunoreactivities for caspase-3 in peri-glandular cells. (B) Melatonin group showing few immunoreactivity for caspase-3 in interstitial cells. (C, D) Cisplatin group showing high immunoreactivity for caspase-3 in the cytoplasm of glandular acini with cytoplasmic expression in invading cellular mass. (E) Cisplatin+melatonin showing minimal immunoreactivity for caspase-3 in peri-glandular cells. Image magnification: 400x; thick arrow: positive glandular epithelial cells; thin arrow: positive cellular mass and interstitial cells. (F) Quantitative analysis of caspase-3 immunostaining (% area). (G) Gene expression of α TGF-β. Data were expressed in mean±SD. !!! indicates p < 0.001 significant versus control rats; ~~~ indicates p < 0.001 significant versus cisplatin group. IHC: immunohistochemistry; TGF-β: transforming growth factor beta; SMG: submandibular gland; Bax: Bcl-2-associated X protein
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Cisplatin in cancer therapy: molecular mechanisms of action. Dasari S, Tchounwou PB. Eur J Pharmacol. 2014;740:364–378. - PMC - PubMed
    1. Phase I/II trial of chemotherapy with docetaxel, cisplatin, and S-1 for unresectable advanced squamous cell carcinoma of the esophagus. Ojima T, Nakamura M, Nakamori M, et al. Oncology. 2018;95:116–120. - PubMed
    1. Histological and immunohistochemical study to evaluate the effects of chamomile versus green tea extracts on the salivary glands of methotrexate treated male albino rats (Doctoral dissertation, Fayoum University) Abdulmonem AM. https://www.fayoum.edu.eg/english/Med/Histology/pdf/DrAlshimaaPhdE.pdf Nat Sci. 2020;18:74–95.
    1. Amburana cearensis leaf extract protects against cisplatin-induced ovarian damage through regulation of p-PTEN and p-Akt proteins in mice. Gouveia BB, Barberino RS, Menezes VG, et al. Iran J Basic Med Sci. 2022;25:683–689. - PMC - PubMed
    1. The effect of different routes of injection of bone marrow mesenchymal stem cells on parotid glands of rats receiving cisplatin: a comparative study. Hany E, Sobh MA, ElKhier MT, ElSabaa HM, Zaher AR. Int J Stem Cells. 2017;10:169–178. - PMC - PubMed

LinkOut - more resources