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Anti-apoptotic and antioxidant effects of melatonin protect spleen of whole body γ-irradiated male Sprague-dawley rats

Published on Wednesday, 22 October 2025

Abstract

Background: Spleen is the largest lymphatic organ that is seriously affected during irradiation. Radiation exposure reduces both of spleen size and weight; that in turn decreases the numbers of immune cells. Melatonin is an effective free radicals scavenger. Therefore, this study aimed to evaluate the effects of melatonin on both blood and spleen of whole body γ-irradiated male Sprague dawley rats.

Materials and methods: Animals were intraperitoneally injected with 100 mg/kg melatonin prior to radiation exposure by 30 minutes. Experimental groups were group I: control rats, group II: irradiated rats, group III: melatonin administrated unirradiated rats and group IV: melatonin administrated irradiated rats. Blood and spleen samples were collected 24 hours post irradiation for biochemical, immunological and blastogenesis measurements. Apoptosis, pro- and anti-apoptotic proteins of spleen cells were measured by flow cytometry techniques.

Results: Melatonin significantly upregulated the activities of superoxide dismutase (SOD), glutathione (GSH) and catalase (CAT); and reduced malondialdehyde (MDA). It down regulated the expression of pro-apoptotic proteins (p53, Bax, caspase-3 and caspase-8) and up regulated the expression of anti-apoptotic protein Bcl-2 in spleen cells; that in turn reduced the radiation-induced apoptosis. Levels of pro-inflammatory cytokines (TNF-α, IFN-γ and IL-1β) were significantly reduced in group IV. Blastogenesis assay showed that melatonin protects PBMC and spleen B lymphocytes and stabilized their proliferation.

Conclusion: Melatonin administration prior to whole body γ-radiation successfully protected rat's spleen from the consequences of radiation exposure. This was due to its free radicle scavenger nature, its reduction of lipid peroxidation and its anti-apoptotic effects.

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- The Di Bella Method (A Fixed Part - Melatonin tablets. From 30-40mg/day up to 200mg/day and more orally in patients with advanced stage of cancer disease and/or patients without respond to traditional treatments);

- Melatonin use in cancer patients have started in 1974, when melatonin prepared according to Prof. Di Bella’s formulation [...]. For 11 days was administered to the patient, admitted to the general medical ward at the Maggiore-Pizzardi Hospital in Bologna, very slowly (over approx. 8 hours) and intravenously administered 1000 mg of melatonin for 11 days. During the course of each day, the patient was intravenously administered 4 saline drips of 500 ml, each containing ten 25 mg bottles of freeze-dried melatonin, lasting 2 hours, totaling 1000 mg per day. No other drug of any kind was administered in order to ascertain the effect of the MLT without interference [...]. From Melatonin with adenosine solubilized in water and stabilized with glycine for oncological treatment - technical preparation, effectivity and clinical findings;

- About Melatonin - In vitro, review and in vivo publications;

- Publication: Melatonin anticancer effects: Review (from Di Bella's Foundation);

- Publication: Key aspects of melatonin physiology: 30 years of research (from Di Bella's Foundation);

- The Di Bella Method (A Fixed Part - Alpha tocopheryl acetate/Vitamin E, approximately 20 grams per day orally);

- Cancer and Vitamin E (analogues and/or derivatives) and cancer - In vitro, review and in vivo publications;

- Solution of retinoids in vitamin E in the Di Bella Method biological multitherapy;

- The Di Bella Method (A Fixed Part - All-Trans Retinoic Acid, Analogues and/or Derivatives - Approximately 60mg per day orally: 40mg per day Beta-Carotene/β-Carotene, 10mg per day ATRA and 10mg per day Axerophthol palmitate);

- All-Trans-Retinoic Acid (ATRA - analogues and/or derivatives) - In vitro, review and in vivo publications;

- The Di Bella Method (A Fixed Part - Cyclophosphamide 50mg tablets and/or Hydroxyurea 500mg tablets, one or two per day);

- The Di Bella Method (A Variable Part - Omega 3 Essential/Unsaturated Fatty Acids. From 1.5 grams up to 3.0 grams per day orally);

The Di Bella's Method: Use of Melatonin since 1974 - together with others chemical compounds - in several Oncological Pathologies:

- A retrospective observational clinical study of triple negative breast cancer cases treated with Di Bella Method: A preliminary data;