Immunomodulatory Effects of Unripe Sapodilla (Manilkara zapota) Fruit Extract Through Inflammatory Cytokine Regulation in Type 1 Diabetic Mice

Abstract

Background:One of the most prevalent metabolic disorderin the world is diabetes mellitus (DM), marked by chronic inflammation of pancreatic β-cells.To alleviate this disease, the response of the immune system can be manipulated to produce anti-inflammatory molecules.This research aims to evaluate the immunomodulatory activities of unripe sapodilla fruit (USF) extract according to the profile of TNF-aand IFN—γ that expressed from CD4 T cells, CD4+CD25+IL-10+, and malondialdehyde (MDA) in type 1 diabetes mellitus (T1DM).Methods:Unripe Sapodilla Fruit (USF) was extracted with water bymaceration and freeze-drying process.Liquid ChromatographyHigh Resolution Mass Spectrophotometry (LC-HRMS)was used to identify the phytochemical content of USF extract. Twenty-five male BALB/c mice were randomized into five groups (n=5). A single high dose (145 mg/kgBW) of streptozotocin (STZ) was administered intraperitoneally to induce T1DM. USF extract was administered orally once a day for 14 days. Blood glucose levels and body weight were assessed once every three days for 15 days. Splenic cells were immunostained with antibodies. The sample underwent a flow cytometric analysis and ANOVA was used to statistically assess all the data with p-value of<0.05 were significant. Results:LC-HRMSrevealed that there were 12 major bioactive compounds in aqueous USF extract, which have activity of anti-diabetic, anti-inflammatory, free radical scavenging, and NF-Kβ inhibitor.The results of USF administration showed that glucose levels in diabetic mice were reduced but not significant. CD4+TNF-a+, CD4+IFN—γ+and MDA expression in diabetic mice decreased after USF administration. Meanwhile, CD4+CD25+IL-10+ expression in diabetic mice increased after USF administration.Conclusion: USF extract acts as an immunomodulator by regulating inflammatory cytokines in a non-dosedependent manner. The ability to regulate inflammatory cytokines is shown by enhancing IL-10 as an anti-inflammatory cytokine to suppress TNF-α and IFN-γ. We also show a decrease in MDA expression as a result of USF's antioxidant activity, which aids in the suppression of inflammatory cytokines in diabetic mice.