SHINTA, Prima Ardinas (2025) Pengaruh Nanopartikel PLGA Sambiloto (Andrographis paniculata) Terhadap Ekspresi mRNA dan Protein IL-4, IL-10 pada Sel Raw 264.7 yang Diinduksi Lipopolisakarida. Masters thesis, Universitas Jenderal Soedirman.

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Abstract

Effect of PLGA Nanoparticles of Ethanolic Extract of Andrographis paniculata on mRNA Expression and Protein Levels of IL-4 and IL-10 in LPS-Induced RAW 264.7 Cells Background: Chronic inflammation contributes to various degenerative and autoimmune diseases. Interleukin-4 (IL-4) and Interleukin-10 (IL-10) are anti-inflammatory cytokines that play important roles in immune regulation, but their effectiveness may decrease under inflammatory conditions. Andrographis paniculata (sambiloto) is known for its anti-inflammatory activity; however, the bioavailability of its main active compound, andrographolide, is relatively low. Nanoparticle formulation using poly lactic-co-glycolic acid (PLGA) has the potential to enhance its effectiveness. Methods: This in vitro study used RAW 264.7 cells induced with lipopolysaccharide (LPS) at 100 ng/mL. Cells were divided into nine groups: media control (A), LPS control (B), sambiloto nanoparticles at 200 ppm (C), 300 ppm (D), 400 ppm (E), 500 ppm (F), and 600 ppm (G), ethanolic sambiloto extract 500 ppm (H), and blank nanoparticles (I). Treatments were administered for 24 hours. IL-4 and IL-10 protein levels were measured using enzyme-linked immunosorbent assay (ELISA), while mRNA expression was assessed using RT-qPCR. Normality was tested with Shapiro–Wilk. Non-normally distributed IL-4 data were analyzed using Kruskal–Wallis followed by Dunn’s post hoc test, while normally distributed IL-10 data were analyzed using ANOVA followed by Tukey’s post hoc test. Results: PLGA nanoparticles of sambiloto extract significantly increased both IL-4 and IL-10 levels and mRNA expression (p<0.05). The highest IL-4 and IL-10 levels were observed at 500 and 600 ppm. IL-10 mRNA expression showed significant upregulation compared to controls. Post hoc analysis revealed significant differences between treatment groups and media control, LPS control, and blank nanoparticles. Conclusion: Administration of PLGA-encapsulated ethanolic extract of Andrographis paniculata nanoparticles enhanced the mRNA expression and protein levels of IL-4 and IL-10 in LPS-induced RAW 264.7 cells, with the strongest effect observed at 500 ppm. These results indicate the potential role of PLGA-based Andrographis paniculata nanoparticles in modulating inflammatory responses.

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