Shahrekord University of Medical Sciences Journal of Herbmed Pharmacology 2345-5004 14 4 2025 10 01 In vitro and in vivo evaluation of Ficus religiosa stem bark extracts, including antioxidant, anti-inflammatory, antimicrobial, antidiarrheal, and antipyretic activities 426 434 10.34172/jhp.2025.52927 EN Samiha Mehnaz https://orcid.org/0009-0008-9220-9675 A.H.M. Nazmul Hasan https://orcid.org/0000-0001-6029-2728 Sanjeeda Ahmed https://orcid.org/0009-0001-4791-4788 Md. Khokon Miah Akanda https://orcid.org/0000-0002-1916-4553 Md. Hamiduzzaman https://orcid.org/0000-0001-8894-8426 Journal Article 10.34172/jhp.2025.52927 2025 01 25 2025 07 16 Introduction: Ficus religiosa has traditionally been used for skin ailments, menstrual disorders, infections, and inflammatory conditions. This study aimed to explore its antioxidant capacity, anti-microbial activity, thrombolytic potential, anti-inflammatory effects, and antipyretic activity. Methods: Antioxidant potential of methanolic and n-hexane extracts was assessed through total phenol, flavonoid, 2,2-diphenyl-1-picrylhydrazyl (DPPH), and nitric oxide (NO) scavenging assays. Anti-microbial activity was tested on selected microbes, cytotoxicity on brine shrimp, and anti-inflammatory properties through membrane-stabilizing potential. Analgesic, antidiarrheal and antipyretic effects were evaluated in mice. Results: The methanol extract had higher total flavonoid and antioxidant capacity content than the n-hexane extract. Both extracts showed substantial total phenolic, flavonoid, and antioxidant content. Methanol and n-hexane extracts showed free radical scavenging capacity against DPPH (IC50: 24.524 μg/mL and 319.669 μg/mL) and NO (IC50: 463.45 μg/mL and 69.45 μg/mL). n-Hexane extract exhibited antimicrobial efficacy against all tested pathogens. At 250 µg/disc, there was a 7 mm zone of inhibition against Pseudomonas spp. and Candida albicans and an 8 mm zone of inhibition against Bacillus subtilis, Escherichia coli, and Streptococcus aureus. Methanol extract had greater anti-inflammatory and thrombolytic activity than n-hexane extract. Maximum antidiarrheal activity was shown by methanol extract (83.48% defecation inhibition, P˂0.001), whereas n-hexane extract demonstrated notable peripheral analgesic activity (72.55% writhing inhibition, P˂0.01). Conclusion: The extract of F. religiosa stem bark has significant therapeutic potential, which might be explored to develop natural remedies for inflammation, microbial infections, and oxidative stress-related disorders. Further study is needed to evaluate its isolated bioactive components.