Cytoprotective And Anti-Inflammatory Effects Of Manilkara Zapota Ethanolic Flower Extract Against H₂O₂-Induced Oxidative Stress In Raw 264.7 Macrophages

Abstract

Background:

Oxidative stress plays a crucial role in the initiation of inflammation and cellular damage. Hydrogen peroxide (H₂O₂) is commonly used to induce oxidative stress in vitro. The present study evaluates the cytoprotective, antioxidant, and anti-inflammatory properties of the ethanolic extract of Manilkara zapota flowers (SFE) on H₂O₂-induced RAW 264.7 macrophage cells.

Methods:

Sapota flowers were collected, shade-dried, and ground into powder. Ethanolic extracts were prepared and tested for cytoprotective and anti-inflammatory activities using MTT assay, nitric oxide (NO) and prostaglandin E2 (PGE2) quantification, DCFDA staining for reactive oxygen species (ROS) assessment, and gene expression analysis via real-time PCR. Morphological changes were observed using phase contrast microscopy.

Results:

SFE was non-toxic up to 200 µg/ml and significantly improved cell viability in H₂O₂-treated cells. SFE effectively reduced NO, PGE2, and ROS levels in a dose-dependent manner. Morphological analysis confirmed the protective effect of SFE against oxidative damage. Gene expression studies showed that SFE downregulated pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-8, Cox-2a) and moderated IL-10 levels, indicating its anti-inflammatory potential.

Conclusion:

The ethanolic extract of Manilkara zapota flowers exhibits strong cytoprotective, antioxidant, and anti-inflammatory activities, suggesting its potential as a therapeutic agent against oxidative stress-related inflammatory conditions.