Aktivitas Imunomodulator Ekstrak Metanol dan Fraksi buah Talok (Muntingia calabura L.) pada Sel RAW 264.7

Tanti Azizah Sujono, Ika Trisharyanti Dian Kusumowati, Rima Munawaroh

Abstract

Buah talok (Muntingia calabura L) secara empiris telah diketahui mempunyai banyak manfaat bagi kesehatan, diantaranya adalah sebagai imunomodulator. Namun mekanisme aksinya sebagai imunomodulator belum diketahui secara pasti. Penelitian ini bertujuan untuk mengevaluasi efek imunomodulator ekstrak metanolik dan fraksi buah talok pada sel makrofag RAW 264.7. Sel RAW 264.7 diberi perlakuan dengan ekstrak metanolik buah talok (EMBT), fraksi heksan buah talok (FHBT), fraksi diklorometana buah talok (FDBT), fraksi etil asetat buah talok (FEABT) serta kelompok kontrol sel (KS), kontrol lipopolisakarida (LPS) dan deksametason. Viabilitas sel diukur dengan metode MTT assay dilanjutkan isolasi RNA, pembuatan cDNA dan PCR. Level ekspresi mRNA dari gen-gen seperti toll like receptor 4 (TLR4), interferon (IFN)-ɣ, interleukin (IL)-6, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α pada sel RAW 264.7 yang diinduksi lipopolisakarida (LPS) dianalisis dengan metode reverse transcription-PCR. Hasil penelitian menunjukkan bahwa deksametason menghambat ekspresi iNOS, IL-6, TNF-α, TLR4, IFN- ɣ dan NF-kB dibandingkan dengan kontrol sel dan LPS. Hal ini mengindikasikan bahwa deksametason mempunyai aktivitas antiinflamasi. Ekstrak metanolik (EMBT) dan fraksi buah talok (FHBT, FDBT dan FEABT) menurunkan ekspresi gen iNOS, TNF-α, IL-6, IFN- ɣ dan NF-kB jika dibandingkan dengan kontrol sel dan LPS, hal ini menunjukkan adanya aktivitas antiinflamasi. Ekstrak metanolik dan fraksi buah talok mempunyai  aktivitas imunomodulator melalui penghambatan inflamasi dengan menurunkan ekspresi gen iNOS, TNF-α, IL-6, IFN- ɣ dan NF-kB pada sel RAW 264.7 yang diinduksi LPS. 

Keywords

Muntingia calabura L; MTT assay; sel RAW 264.7; Reverse Transcription-Polymerase Chain Reaction (RT-PCR)

Full Text:

PDF

References

Ahmed, A. U. (2011). An overview of inflammation: Mechanism and consequences. Frontiers of Biology in China, 6(4), 274–281.

Auliafendri, N., Rosidah, Yuandani, Suryani, S., and Satria, D. (2019). The immunomodulatory activities of Picria Fel-Terrae lour herbs towards RAW 264.7 cells. Open Access Macedonian Journal of Medical Sciences, 7(1), 24–28.

Baratawidjaja, K. G., and Rengganis, I. (2016). Imunologi Dasar (11th ed.). Badan Penerbit Fakultas Kedokteran Universitas Indonesia.

Chuang, T. Y., Cheng, A. J., Chen, I. T., Lan, T. Y., Huang, I. H., Shiau, C. W., Hsu, C. L., Liu, Y. W., Chang, Z. F., Tseng, P. H., and Kuo, J. C. (2017). Suppression of LPS-induced inflammatory responses by the hydroxyl groups of dexamethasone. Oncotarget, 8(30), 49735–49748.

Darmawan, K. H., Martien, R., Erlangga, N. D., Sitohang, S. M., and Pambudi, H. (2017). Utilization Of Nano Ethanolic Extract Combination Chamber Bitter (Phyllanthus Niruri L.) And Garlic (Allium Sativum L.) As A Natural Immunomodulator In Nanoherbal Development, In Silico And In Vitro Study. JPSCR : Journal of Pharmaceutical Science and Clinical Research, 2(02), 110.

Dewi, K., Widyarto, B., Erawijantari, P., and Widowati, W. (2015). In vitro study of Myristica fragrans seed (Nutmeg) ethanolic extract and quercetin compound as anti-inflammatory agent. International Journal of Research in Medical Sciences, 3(9), 2303–2310.

Fard, M., Arulselvan, P., Karthivashan, G., Adam, S., and Fakurazi, S. (2015). Bioactive extract from moringa oleifera inhibits the pro-inflammatory mediators in lipopolysaccharide stimulated macrophages. Pharmacognosy Magazine, 11(44), 556.

Fitri, D., Kiromah, N. Z. W., and Widiastuti, T. C. (2020). Formulasi Dan Karakterisasi Nanopartikel Ekstrak Etanol Daun Salam (Syzygium polyanthum) Pada Berbagai Variasi Komposisi Kitosan Dengan Metode Gelasi Ionik. JPSCR: Journal of Pharmaceutical Science and Clinical Research, 5(1), 61.

García-Lafuente, A., Guillamón, E., Villares, A., Rostagno, M. A., and Martínez, J. A. (2009). Flavonoids as anti-inflammatory agents: Implications in cancer and cardiovascular disease. Inflammation Research, 58(9), 537–552.

González, R., Ballester, I., López-Posadas, R., Suárez, M. D., Zarzuelo, A., Martínez-Augustin, O., and Sánchez de Medina, F. (2011). Effects of flavonoids and other polyphenols on inflammation. Critical Reviews in Food Science and Nutrition, 51(4), 331–362.

Grigore, A. (2017). Plant Phenolic Compounds as Immunomodulatory Agent. In Phenolic Compound-Biological Activity (pp. 75–88).

Hikmah, N., and Dewanti, I. D. A. R. (2011). Peran Toll Like Receptors (TLRs) pada Innate Immunity (the Role of Toll Like REceptors to Innate Immunity). Stomatognatic, 8(1), 21–26.

Joo, T., Sowndhararajan, K., Hong, S., Lee, J., Park, S. Y., Kim, S., and Jhoo, J. W. (2014). Inhibition of nitric oxide production in LPS-stimulated RAW 264.7 cells by stem bark of Ulmus pumila L. Saudi Journal of Biological Sciences, 21(5), 427–435.

Kim, T., Song, B., Cho, K. S., and Lee, I. S. (2020). Therapeutic potential of volatile terpenes and terpenoids from forests for inflammatory diseases. International Journal of Molecular Sciences, 21(6).

Kwon, D. H., Cheon, J. M., Choi, E.-O., Jeong, J. W., Lee, K. W., Kim, K. Y., Kim, S. G., Kim, S., Hong, S. H., Park, C., Hwang, H.-J., and Choi, Y. H. (2016). The Immunomodulatory Activity of Mori folium, the Leaf of Morus alba L., in RAW 264.7 Macrophages In Vitro . Journal of Cancer Prevention, 21(3), 144–151.

Martins, G. R., Gelaleti, G. B., Moschetta, M. G., Maschio-Signorini, L. B., and De Campos Zuccari, D. A. P. (2016). Proinflammatory and Anti-Inflammatory Cytokines Mediated by NF-κB Factor as Prognostic Markers in Mammary Tumors. Mediators of Inflammation, 2016.

Muniandy, K., Gothai, S., Badran, K. M. H., Kumar, S. S., Esa, N. M., and Arulselvan, P. (2018). Suppression of proinflammatory cytokines and mediators in LPS-Induced RAW 264.7 macrophages by stem extract of alternanthera sessilis via the inhibition of the NF-κB pathway. Journal of Immunology Research, 2018.

Murray, P. J., and Wynn, T. A. (2011). Protective and pathogenic functions of macrophage subsets. Nature Reviews Immunology, 11(11), 723–737.

Okamoto, I., Iwaki, K., Koya-Miyata, S., Tanimoto, T., Kohno, K., Ikeda, M., and Kurimoto, M. (2002). The flavonoid kaempferol suppresses the graft-versus-host reaction by inhibiting type 1 cytokine production and CD8+ T cell engraftment. Clinical Immunology, 103(2), 132–144.

Patel, S., and Vajdy, M. (2015). Induction of cellular and molecular immunomodulatory pathways by vitamin A and flavonoids. Expert Opinion on Biological Therapy, 15(10), 0.

Pereira, G. A., Arruda, H. S., de Morais, D. R., Eberlin, M. N., and Pastore, G. M. (2018). Carbohydrates, volatile and phenolic compounds composition, and antioxidant activity of calabura (Muntingia calabura L.) fruit. Food Research International, 108(March), 264–273.

Prakash, V. (2017). Terpenoids as source of anti-inflammatory compounds. Asian Journal of Pharmaceutical and Clinical Research, 10(3), 68–76.

Prakosa, A. P. (2014). Imunomodulasi Ekstrak Buah Talok (Muntingia calabura Linn) terhadap Makrofag Peritoneum Tikus Wistar Jantan yang Diinduksi Actinobacillus actionycetemcomitans. Universitas Jendral Soedirman.

Preethi, K., Premasudha, P., and Keerthana, K. (2012). Anti-inflammatory activity of Muntingia calabura fruits. Pharmacognosy Journal, 4(30), 51–56.

Preethi, K., Vijayalakshmi, N., Shamna, R., and Sasikumar, J. M. (2010). In Vitro Antioxidant Activity of Extracts from Fruits of Muntingia calabura Linn . from India. Pharmacognosy Journal, 2(14), 11–18.

Rosidah, Yuandani, Widjaja, S. S., Lubis, M. F., and Satria, D. (2019). The Immunomodulatory Activities of Saurauia vulcani Korth Leaves towards RAW 264 . 7 cell. International Summit on Science Technology and Humanity (ISETH), 586–593.

Sarimanah, J., Ketut Adnyana, I., Sukandar, E. Y., and Kurniati, N. F. (2017). The antirheumatic activity of Muntingia calabura L. Leaves ethanol extract and its fraction. Asian Journal of Pharmaceutical and Clinical Research, 10(1), 84–86.

Sasmito, E. (2017). Imunomodulator Bahan Alami. Rapha Publising Penerbit ANDI Yogyakarta.

Senet, M. R. M., Parwata, I. M. O. A., and Sudiarta, I. W. (2012). Kandungan Total Fenol dan Flavonoid dari Buah Kersen (Muntingia calabura) serta Aktivitas Antioksidannya. Jurnal Kimia, 11(2), 187–193.

Siddiqua, A., Premakumari, K. B., Sultana, R., Vithya, and Savitha. (2010). Antioxidant activity and estimation of total phenolic content of Muntingia calabura by colorimetry. International Journal of ChemTech Research, 2(1), 205–208.

Sujono, T. A., Kusumowati, I. T. D., and Munawaroh, R. (2020). Immunomodulatory Activity of Muntingia calabura L Fruits using Carbon Clearance Assay and Their Total Flavonoid and Phenolic Contents. Asian Journal of Pharmaceutical and Clinical Research, 13(2), 140–145.

Tungmunnithum, D., Thongboonyou, A., Pholboon, A., and Yangsabai, A. (2018). Flavonoids and Other Phenolic Compounds from Medicinal Plants for Pharmaceutical and Medical Aspects: An Overview. Medicines, 5(3), 93.

Yahfoufi, N., Alsadi, N., Jambi, M., and Matar, C. (2018). The immunomodulatory and anti-inflammatory role of polyphenols. Nutrients, 10(11), 1–23.

Yanti, Pramudito, T. E., Nuriasari, N., and Juliana, K. (2011). Lemon Pepper Fruit Extract (Zanthoxylum acanthopodium DC.) Suppresses the Expression of Inflammatory Mediators in Lipopolysaccharide-Induced Macrophages In Vitro. American Journal of Biochemistry and Biotechnology, 7(4), 190–195.

Refbacks

  • There are currently no refbacks.