IMMUNOSTIMULATORY ACTIVITY OF SNAKE FRUIT (Salacca edulis Reinw.) CULTIVAR PONDOH HITAM EXTRACT ON THE ACTIVATION OF MACROPHAGES IN VITRO AND IN VIVO

SRI WIJANARTI, Prof. Dr. Ir. Eni Harmayani, M.Sc.; Dr. Ir. Supriyadi, M.Sc.

2015 | Tesis | S2 Ilmu dan Teknologi Pangan

Salak (Salacca edulis Reinw.) kultivar pondoh hitam adalah buah tropis yang tumbuh di Indonesia. Salak dikonsumsi segar atau olahan namun menghasilkan kulit sebagai limbah. Daging dan kulit salak mengandung polisakarida yang tinggi seperti pektin dan serat makanan. Selain itu, salak pondoh juga mengandung senyawa fenolik yang tinggi. Oleh karena itu, salak pondoh merupakan kandidat immunostimulator yang potensial. Namun, efek imunologi dari salak pondoh belum dilaporkan. Penelitian ini bertujuan untuk mempelajari aktivitas imunostimulatori ekstrak daging salak pondoh hitam (EDSPH) dan esktrak kulit salak pondoh hitam (EKSPH) pada aktivasi makrofag dievaluasi. EDSPH dan EKSPH diekstraksi dari daging buah dan kulit salak menggunakan air selama 15 jam pada suhu 4C. EDSPH digunakan untuk menstimulasi aktivitas fagositosis, produksi sitokin, dan ekspresi gen secara in vitro menggunakan sel makrofag J774.1 dan in vivo menggunakan tikus BALB/c. EKSPH digunakan untuk merangsang aktivitas fagositosis dan produksi sitokin in vitro. Analisis imunoblot dilakukan untuk mengevaluasi mekanisme aktivasi EKSPH pada makrofag. Hasil penelitian menunjukkan bahwa EDSPH dan EKSPH memiliki aktivitas imunostimulatori. EDSPH meningkatkan aktivitas fagositosis pada sel J774.1. Produksi sitokin seperti TNF-Alpha dan IL-6 serta ekspresi gennya ditingkatkan olehEDSPH in vitro dan in vivo. EKSPH meningkatkan aktivitas fagositosis, produksi TNF-Alpha dan IL-6, serta tingkat ekspresi gennya. Analisis imunoblot menunjukkan bahwa EKSPH mengaktifkan makrofag melalui jalur sinyal Nuclear Factor (NF)-kappa B dan dua anggota jalur sinyal Mitogen-Activated Protein Kinase yaitu JNK dan p38. Keseluruhan temuan menunjukkan bahwa EDSPH dan EKSPH meningkatkan sistem kekebalan tubuh dengan cara mengaktifkan makrofag.

Snake fruit (Salacca edulis Reinw) cultivar pondoh hitam is a tropical fruit produced in Indonesia. Snake fruit is consumed freshly or processed and left the peel as waste. Snake fruit flesh and the peel, contain high polisaccharides such as pectin, dietary fiber, and high phenolic compound. Therefore, snake fruit is a potential immunostimulator candidates but the immunological effect of snake fruit has not been reported. In the present study, immunostimulatory activity of snake fruit flesh extract (SFFE) and snake fruit peel extract (SFPE) on macrophages activation was evaluated. SFFE and SFPE were prepared by extracting from snake fruit flesh and peel powder in water for 15 h at 4C. SFFE was used for phagocytosis activity, cytokines production, and gene expression stimulation in vitro using murine macrophage-like J774.1 cells and in vivo using BALB/c mice. SFPE was used for stimulating phagocytosis activity and cytokines production in vitro. Then, immunoblot analysis was done to evaluate underlying mechanism of SFPE activation on macrophages. The results showed that SFFE and SFPE exhibited immunostimulatory activities. SFFE increased phagocytosis activity on J774.1 cells. Cytokines such as TNF-Alfa and IL-6 production as well as their genes expression level were elevated by SFFE in vitro and in vivo. SFPE increased phagocytosis activity, TNF-Alfa and IL-6 production, and their gene expression level. Immunoblot analysis revealed that SFPE activated macrophages through Nuclear Factor (NF)-kappa B signaling cascade and two member of Mitogen-Activated Protein Kinases signaling cascade which were JNK and p38. Overall findings suggest that SFFE and SFPE have a possible beneficial effects to promote the body health through activating macrophages.

Kata Kunci : salak, imunostimulatori, fagositosis, sitokin, sel J774.1, JNK, NF-kappa B