Possible hemoglobin enhancing effect of phytol in methotrexate-induced folate deficient Swiss albino mice: In vivo and in silico studies

Abstract

The diterpenoid phytol is evidently acting against anemia in experimental animals. However, the molecular mechanisms behind this issue are yet to be discovered. This study aimed to evaluate phytol's effect on methotrexate-induced folate-deficient animals through in vivo and in silico studies. For this, a total of thirty adult male Swiss albino mice were randomly divided into six different groups, namely the normal control (vehicle), the negative control (folate deficiency inducer, methotrexate 3 ​mg/kg), the standard (folic acid 1.5 ​mg/kg), two test groups comprising phytol 25 and 50 ​mg/kg, and a combined group composed of the standard and highest test doses of phytol. Except for the vehicle, all groups were treated with methotrexate for the first 3 days (once/day) to induce folate deficiency. Then followed by the respective treatment once a day for 3 days. Hemoglobin (Hb) level was measured from the peripheral blood (by tail cutting) on days 1st (before treatment), 4th (after methotrexate treatment), and 7th (after treatment). On the other hand, the computational studies were performed by PyMol, PyRex, Discovery Studio, and other complementary tools. Findings suggest that phytol significantly (p ​< ​0.05) augmented Hb levels that are altered by methotrexate-induced reduction of Hb levels in animals dose-dependently. The combination also augmented Hb levels in animals; however, its effect was slightly lower than the individual groups (standard and test). In the in silico study, phytol showed good binding capacity (binding energy: −7.0 ​kcal/mol) with dihydrofolate reductase (DHFR). In conclusion, phytol may act against folate deficiency by altering methotrexate's impacts in animals, possibly through interacting with DHFR. Further validated research is necessary to develop phytol as an anti-anemia drug in the near future.

Keywords

Hemoglobin / Folate deficiency / Phytol / Methotrexate / Folic acid

Data availability

This published article contains the data collected and studied during this investigation. Information will be provided upon valid request.

Ethics approval

This study was granted by the Ethical Committee of Bangabandhu Sheikh Mujibur Rahman Science and Technology University [#BSMRSTU-RC/PHR-2023-33].

Funding

Researcher supporting project number (PHR-2023-33), Bangabandhu Sheikh Mujibur Rahman Science and Technology University, Gopalganj 8100, Bangladesh.

CRediT authorship contribution statement

Muhammad Torequl Islam: Conceptualization. Raihan Chowdhury: Investigation, Data curation. Md Sakib Al Hasan: Investigation, Formal analysis. Salehin Sheikh: Methodology, Data curation. Md Shimul Bhuia: Methodology, Formal analysis. Sumaya Akter Bithi: Software. Most Israt Jahan Oni: Writing - original draft. Mehedi Hasan Bappi: Resources, Methodology. Siddique Akber Ansari: Supervision. Elaine C.P. Lucetti: Validation, Methodology. Catarina M. Tahim: Supervision, Methodology. Henrique Douglas Melo Coutinho: Project administration. Irfan Aamer Ansari: Supervision.

Declaration of competing interest

The authors declare that they have no known competing financialinterestsor personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgment

We are thankful to the researcher supporting project (PHR-2023-33), gabandhu Sheikh Mujibur Rahman Science and Technology University, Gopalganj 8100, Bangladesh.

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