Synergistic Anti Inflammatory Effects of Galangin and Piperine in a Bleomycin Induced Pulmonary Fibrosis Model in Mice
K. Vanitha Sree *
Administrative Office, P.V. Narsimha Rao Telangana Veterinary University, Hyderabad, India.
M. Usha Rani
College of Veterinary Science, Mamnoor, India.
A. Gopala Reddy
College of Veterinary Science, Rajendra Nagar, India.
B. Rajender
College of Veterinary Science, Rajendra Nagar, India.
B. Swathi
College of Veterinary Science, Mamnoor, India.
D. Durga Veera Hanuman
College of Veterinary Science, Rajendra Nagar, India.
K V Venkata Rao
College of Veterinary Science, Garividi, India.
D. Bhaskar
College of Veterinary Science, Rajendra Nagar, India.
*Author to whom correspondence should be addressed.
Abstract
Aims: To evaluate the anti‑inflammatory efficacy of galangin and piperine administered individually and in combination on broncho‑alveolar lavage fluid (BALF) cellularity and lung cytokine profiles in a bleomycin‑induced pulmonary fibrosis model in mice.
Study Design: Randomized controlled animal experiment.
Place and Duration of Study: Department of Veterinary Pharmacology and Toxicology, College of Veterinary Science, Hyderabad, India, from June 2021 to August 2021.
Methodology: Fifty male C57BL/6 mice (6–7 weeks) were acclimatized for 15 days and then allocated to seven groups (n = 6 for analysis). Group 1 received saline only; Group 2 received a single oropharyngeal dose of bleomycin (1.5 U kg⁻¹). Groups 3 and 4 received galangin (5 mg kg⁻¹ day⁻¹) and piperine (50 mg kg⁻¹ day⁻¹) alone respectively. Groups 5 and 6 received bleomycin plus galangin or piperine, respectively, at the same doses following day of bleomycin. Group 7 received bleomycin plus doses of galangin (2.5 mg kg⁻¹ day⁻¹) and piperine (25 mg kg⁻¹ day⁻¹). After 21 days, BALF was collected for total and differential cell counts; lung homogenates were analyzed by ELISA for TNF‑α, NF‑κB, IL‑6, and IL‑10. Data were expressed as mean ± SE and compared by one‑way ANOVA with Tukey’s post‑hoc test (P < 0.05).
Results: Bleomycin significantly increased BALF total cells (20.33 ± 0.33 × 10⁵ vs 9.53 ± 0.58 × 10⁵ in controls) and neutrophils (4.61 ± 0.24 × 10⁵ vs 1.39 ± 0.14 × 10⁵; P < 0.05). Galangin and piperine monotherapy reduced these counts by ~26 % and ~29 %, respectively, while combination therapy achieved a 34 % reduction (13.48 ± 0.12 × 10⁵). Bleomycin also elevated TNF‑α, NF‑κB, and IL‑6 (5‑ to 6‑fold) and suppressed IL‑10 (60 % decrease). Combination treatment lowered TNF‑α to 64.37 ± 1.13 pg mg⁻¹ (vs 145.70 ± 1.94 pg mg⁻¹ in bleomycin group) and NF‑κB to 77.88 ± 2.84 pg mg⁻¹ (vs 173.53 ± 2.14 pg mg⁻¹), while restoring IL‑10 toward baseline (37.25 ± 1.39 pg mg⁻¹ vs 43.73 ± 1.53 pg mg⁻¹ in controls) (all P < 0.05).
Conclusion: Galangin and piperine significantly attenuate bleomycin‑induced pulmonary inflammation by reducing leukocyte infiltration and re‑balancing pro‑ and anti‑inflammatory cytokines. Combination therapy displayed superior efficacy, suggesting a synergistic interaction and supporting the potential use of these phytocompounds as adjunct treatments for inflammatory lung diseases.
Keywords: Lung injury, cytokines, bleomycin, galangin, piperine