GARCINIA COWA ROXB. ETHANOL EXTRACT INHIBITS INFLAMMATION IN LPS-INDUCED RAW 264.7 MACROPHAGES

IRENE PUSPA DEWI; FATMA WAHYUNI; YUFRI ALDI; DACHRIYANUS

doi:10.22159/ijap.2023.v15s1.01

Authors

IRENE PUSPA DEWI Prayoga Pharmacy Academy, Jalan Jenderal Sudirman, Padang, Indonesia, Faculty of Pharmacy, Andalas University, Padang, Indonesia
FATMA WAHYUNI Faculty of Pharmacy, Andalas University, Padang, Indonesia
YUFRI ALDI Faculty of Pharmacy, Andalas University, Padang, Indonesia
DACHRIYANUS Faculty of Pharmacy, Andalas University, Padang, Indonesia

DOI:

https://doi.org/10.22159/ijap.2023.v15s1.01

Keywords:

Garcinia cowa Roxb, LPS-induced RAW 264.7 macrophages, Inflammation

Abstract

Objective: The purpose of this study was to examine the effect of Garcinia cowa Roxb. Ethanol (EGC) extract in LPS-induced Raw 264.7 macrophages by observing the release of Tumor Necrosis Factor (TNF) and Interleukin-6 (IL-6).

Methods: Using the MTT method, a cell viability assay was performed to observe the cytotoxic effect on Raw 264.7 macrophages. For 24 h, Raw 264.7 macrophages were incubated with various EGC concentrations (100, 50, 10, 1 and 0.1 µg/ml). The medium was taken out after 48 h of incubation, and 100 µl of MTT 0.5 mg/ml was then added. 100 µl DMSO was used to dissolve the crystals and absorbance was measured using a microplate reader. To investigate the activity of EGC to LPS-induced Raw 264.7 macrophages, the ELISA method was used. Supernatant was obtained after treating Raw 264.7 macrophages with complete medium, EGC samples, and LPS (10 g/ml) for 24 h. IL-6 and TNF-α levels were assessed using supernatants with ELISA kit.

Results: Cytotoxic effect of EGC to Raw 264.7 macrophages occurred at a concentration of 100 µg/ml with the cell viability value of 59.5%. At a concentration of 50 µg/ml, no cytotoxic effect occurred and the cell viability value was 105.5%. So, the higher concentration of EGC used for further investigation is 50 µg/ml. It was shown that the production of IL 6 was suppressed by EGC at a concentration of 12.5 µg/ml. The inhibition of TNF-α production was only seen at the concentration of 12.5, 25 and 50 µg/ml; there was an increase of TNF-α production.

Conclusion: It can be concluded that EGC can be developed as a natural immunomodulator that can inhibit inflammation by suppressing IL-6 production to prevent immune system disorders.

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References

Sindhu RK, Goyal A, Das J, Neha, Choden S, Kumar P. Immunomodulatory potential of polysaccharides derived from plants and microbes: a narrative review. Carbohydrate Polymer Technologies and Applications. 2021;2. doi: 10.1016/j.carpta.2021.100044.

Zheng T, Gu D, Wang X, Shen X, Yan L, Zhang W. Purification, characterization and immunomodulatory activity of polysaccharides from Leccinum crocipodium (Letellier.) watliag. Int J Biol Macromol. 2020;148:647-56. doi: 10.1016/j.ijbiomac.2020.01.155, PMID 31958555.

Ahmad W, Jantan I, Kumolosasi E, Haque MA, Bukhari SNA. Immunomodulatory effects of Tinospora crispa extract and its major compounds on the immune functions of RAW 264.7 macrophages. Int Immunopharmacol. 2018;60(April):141-51. doi: 10.1016/j.intimp.2018.04.046, PMID 29730557.

Babich O, Sukhikh S, Prosekov A, Asyakina L, Ivanova S. Medicinal plants to strengthen immunity during a pandemic. Pharmaceuticals (Basel). 2020;13(10):1-18. doi: 10.3390/ph13100313, PMID 33076514.

Jantan I, Ahmad W, Bukhari SNA. Plant-derived immunomodulators: an insight on their preclinical evaluation and clinical trials. Front Plant Sci. 2015;6:655. doi: 10.3389/fpls.2015.00655, PMID 26379683.

Wahyuni FS, Stanslas J, Lajis NH, Dachriyanus. Cytotoxicity studies of tetraprelyltoluquinone, a prenilated hydroquinone from Garcina cowa Roxb on H-460, MCF-7 and DU-145. Int J Pharm Pharm Sci. 2015;7(3):60-3.

Wahyuni FS, Triastuti DH, Arifin H. Cytotoxicity study of ethanol extract of the leaves of asam kandis (Garcinia cowa Roxb.) on T47D breast cancer cell line. Phcog J. 2015;7(6):369-71. doi: 10.5530/pj.2015.6.9.

Wahyuni FS, Shaari K, Stanslas J, Lajis NH, Hamidi D. Cytotoxic properties and complete nuclear magnetic resonance assignment of isolated xanthones from the root of Garcinia cowa Roxb. Pharmacogn Mag. 2016;12(Suppl 1):S52-6. doi: 10.4103/0973-1296.176115, PMID 27041859.

Panthong K, Hutadilok Towatana N, Panthong A. Cowaxanthone F, a new tetraoxygenated xanthone, and other anti-inflammatory and antioxidant compounds from Garcinia cowa. Can J Chem. 2009;87(11):1636-40. doi: 10.1139/V09-123.

Wahyuni FS, Shaari K, Stanslas J, Lajis N, Hamidi D. Cytotoxic compounds from the leaves of Garcinia cowa roxb. J App Pharm Sci. 2015;5(2):6-11. doi: 10.7324/JAPS.2015.50202.

Rao RR. Ethnobotany of Meghalaya: medicinal plants used by Khasi and Garo tribes. Econ Bot. 1981;35(1):4-9. doi: 10.1007/BF02859208.

Mahabusarakam W, Chairerk P, Taylor WC. Xanthones from Garcinia cowa Roxb. latex. Phytochemistry. 2005;66(10):1148-53. doi: 10.1016/j.phytochem.2005.02.025, PMID 15924919.

Jabit ML, Wahyuni FS, Khalid R, Israf DA, Shaari K, Lajis NH. Cytotoxic. Pharm Biol. 2009;47(11):1019-26. doi: 10.3109/13880200902973787.

Wahyuni FS, Israf Ali DA, Lajis NH, DD. Anti-inflammatory activity of isolated compounds from the stem bark of Garcinia cowa Roxb. Pharmacogn J. 2016;9(1):55-7. doi: 10.5530/pj.2017.1.10.

Nakatani K, Nakahata N, Arakawa T, Yasuda H, Ohizumi Y. Inhibition of cyclooxygenase and prostaglandin E2 synthesis by γ-mangostin, a xanthone derivative in mangosteen, in C6 rat glioma cells. Biochem Pharmacol. 2002;63(1):73-9. doi: 10.1016/s0006-2952(01)00810-3, PMID 11754876.

Guo C, Bi J, Li X, Lyu J, Liu X, Wu X. Immunomodulation effects of polyphenols from thinned peach treated by different drying methods on RAW264.7 cells through the NF-κB and Nrf2 pathways. Food Chem. 2021;340(2):127931. doi: 10.1016/j.foodchem.2020.127931. PMID 32871358.

Wen L, Huang L, Li Y, Feng Y, Zhang Z, Xu Z. New peptides with immunomodulatory activity identified from rice proteins through peptidomic and in silico analysis. Food Chem. 2021;364(June):130357. doi: 10.1016/j.foodchem.2021.130357. PMID 34174647.

Yang L, Liu J, Xia X, Wong IN, Chung SK, Xu B. Sulfated heteropolysaccharides from Undaria pinnatifida: structural characterization and transcript-metabolite profiling of immunostimulatory effects on RAW264.7 cells. Food Chem X. 2022;13:100251. doi: 10.1016/j.fochx.2022.100251. PMID 35498964.

Toopcham T, Mes JJ, Wichers HJ, Yongsawatdigul J. Immunomodulatory activity of protein hydrolysates derived from Virgibacillus halodenitrificans SK1-3-7 proteinase. Food Chem. 2017;224:320-8. doi: 10.1016/j.foodchem.2016.12.041, PMID 28159274.

Feng D, Ling WH, Duan RD. Lycopene suppresses LPS-induced NO and IL-6 production by inhibiting the activation of ERK, p38MAPK, and NF-κB in macrophages. Inflamm Res. 2010;59(2):115-21. doi: 10.1007/s00011-009-0077-8, PMID 19693648.

Lee TK, Trinh TA, Lee SR, Kim S, So HM, Moon E. Bioactivity-based analysis and chemical characterization of anti-inflammatory compounds from Curcuma zedoaria rhizomes using LPS-stimulated RAW264.7 cells. Bioorg Chem. 2019;82:26-32. doi: 10.1016/j.bioorg.2018.09.027, PMID 30267971.

Lauro R, Irrera N, Eid AH, Bitto A. Could antigen-presenting cells represent a protective element during sars-cov-2 infection in children? Pathogens. 2021;10(4):1-16. doi: 10.3390/pathogens10040476, PMID 33920011.

Novilla A, Djamhuri DS, Nurhayati B, Rihibiha DD, Afifah E, Widowati W. Anti-inflammatory properties of oolong tea (Camellia sinensis) ethanol extract and epigallocatechin gallate in LPS-induced RAW 264.7 cells. Asian Pac J Trop Biomed. 2017;7(11):1005-9. doi: 10.1016/j.apjtb.2017.10.002.

Zelova H, Hosek J. TNF-α signalling and inflammation: interactions between old acquaintances. Inflamm Res. 2013;62(7):641-51. doi: 10.1007/s00011-013-0633-0, PMID 23685857.

Koyama T, Uchida K, Fukushima K, Ohashi Y, Uchiyama K, Inoue G. Elevated levels of TNF-α, IL-1β and IL-6 in the synovial tissue of patients with labral tear: a comparative study with hip osteoarthritis. BMC Musculoskelet Disord. 2021;22(1):33. doi: 10.1186/s12891-020-03888-w, PMID 33407301.

Zhang X, Wang G, Gurley EC, Zhou H. Flavonoid apigenin inhibits lipopolysaccharide-induced inflammatory response through multiple mechanisms in Macrophages. Plos One. 2014;9(9):e107072. doi: 10.1371/journal.pone.0107072, PMID 25192391.