• Yosie Andriani Institute of Marine Biotechnology, University of Malaysia Terengganu, Kuala Nerus, Terengganu, Malaysia.
  • Leni Marlina Institute of Marine Biotechnology, University of Malaysia Terengganu, Kuala Nerus, Terengganu, Malaysia.
  • Habsah Mohamad Institute of Marine Biotechnology, University of Malaysia Terengganu, Kuala Nerus, Terengganu, Malaysia.
  • Hermansyah Amir Educational Chemistry Program, Faculty of Teacher Training and Education, Bengkulu University, Bengkulu, Indonesia.
  • Siti Aisha M Radzi Institute of Marine Biotechnology, University of Malaysia Terengganu, Kuala Nerus, Terengganu, Malaysia.
  • Jasnizat Saidin School of Marine Sciences and Environmental, Universiti Malaysia Terengganu, Kuala Nerus, Malaysia



Haliclona amboinensis, Associated bacteria, Anti-inflammatory, Inducible nitric oxide, Cyclooxygenase-1, Cyclooxygenase-2, Secretory phospholipase A



 Objective: This study aimed to investigate the anti-inflammatory activity of methanol extract and fractions of bacteria associated with sponge (Haliclona amboinensis) and to evaluate their effect in reducing NO production and inhibiting cyclooxygenase-1 (COX-1), cyclooxgenase-2 (COX-2) and secretory phospholipase A2 (sPLA2) activity.

Methods: All bacterial isolates were cultured and supernatants were collected for the extraction of secondary metabolites using diaion HP-20 to obtain methanol extracts. Evaluation of cytotoxicity property was carried out on macrophage cell lines (RAW264.7) by 3-(4,5-dimethylthiazol- 2-yl) 2,5-diphenyl tetrazoliumbromide assay. Anti-inflammatory screening was done by inducible nitric oxide assay on RAW264.7 cell lines with lipopolysaccharide (LPS) stimulation. Dianion HP-20 was used to remove salt content. A selected methanol extract was subjected to further fractionations by C-18 reverse phase and their anti-inflammatory potential was evaluated by COX-1 and COX-2, and sPLA2 enzymatic assay.

Results: Seven methanol extracts showed no cytotoxic property against RAW 264.7 cell line (inhibitory concentration 50% > 30 μg/ml) and selected for anti-inflammatory screening assay. Result showed methanol extract HM 1.2 reduced NO production >80% and it has been selected for phytochemical screening, further fractionations and assay. Phytochemical screening showed alkaloids and terpenoids present in the HM 1.2. The HM 1.2 and its fractions (F1, F2, F1C1, F1C2, F1C3, and F1C4) were proven to inhibit COX-1, COX-2, and sPLA2 activity in the range of 60.516-116.886%, 20.554- 116.457%, and 70.2667-114.8148%, respectively.

Conclusions: This study revealed that bacteria associated with H. amboinensis have produced anti-inflammatory activity via reducing NO production and inhibiting COX-1, COX-2, and sPLA2 activity.



Lubberts E, Van den Berg WB. Cytokines in the Pathogenesis of Rheumatoid Arthritis and Collagen-Induced Arthritis. Madame Curie Bioscience Database. Austin, TX: Landes Bioscience; 2000. p. 2000-13.

Cushnie TP, Cushnie B, Lamb AJ. Alkaloids: An overview of their antibacterial, antibiotic-enhancing and anti-virulence activities. Int J Antimicrob Agents 2014;44(5):377-86.

Mabhiza D, Chitemerere T, Mukanganyama S. Antibacterial properties of alkaloid extracts from Callistemon citrinus and Vernonia adoensis against Staphylococcus aureus and Pseudomonas aeruginosa. Int J Med Chem 2016;2016:6304163.

Li X, Li JR, Chen K, Zhu HL. A functional scaffold in marine alkaloid: An anticancer moiety for human. Curr Med Chem 2013;20(31):3903-22.

Tiong SH, Looi CY, Hazni H, Arya A, Paydar M, Wong WF, et al. Antidiabetic and antioxidant properties of alkaloids from Catharanthus roseus (L.) G. Don. Molecules 2013;18(8):9770-84.

Souto AL, Tavares JF, da Silva MS, Diniz Mde F, de Athayde-Filho PF, Barbosa Filho JM. Anti-inflammatory activity of alkaloids: An update from 2000 to 2010. Molecules 2011;16(10):8515-34.

Prakash V. Terpenoids as source of anti-inflammatory compounds. Asian J Pharm Clin Res 2017;10(3):68-76.

Boobathy S, Soundarapandian P, Subasri V, Vembu N, Gunasundari V. Bioactivities of protein isolated from marine sponge, Sigmadocia fibulatus. Curr Res J Biol Sci 2009;1(3):160-2.

Yadav RN, Agarwala M. Phytochemical analysis of some medicinal plants. J Phytol 2011;3(12):10-4.

Jaganathan SK, Mandal M. Antiproliferative effects of honey and of its polyphenols: A review. J Biomed Biotechnol 2009;2009:830616.

Andriani Y, Ramli NM, Syamsumir DF, Kassim MN, Jaafar J, Azis NA, et al. Phytochemical analysis, antioxidant, antibacterial and cytotoxicity activities of keys and cores part of Pandanus tectorius fruits. Arab J Chem. DOI: 10.1016/j.arabjc.2015.11.003.

Kassim M, Achoui M, Mustafa MR, Mohd MA, Yusoff KM. Ellagic acid, phenolic acids, and flavonoids in Malaysian honey extracts demonstrate in vitro anti-inflammatory activity. Nutr Res 2010;30(9):650-9.

Yi PF, Bi WY, Shen HQ, Wei Q, Zhang LY, Dong HB, et al. Inhibitory effects of sulfated 20(S)-ginsenoside Rh2 on the release of pro-inflammatory mediators in LPS-induced RAW 264.7 cells. Eur J Pharmacol 2013;712(1-3):60-6.

Lau KM, Lai KK, Liu CL, Tam JC, To MH, Kwok HF, et al. Synergistic interaction between astragali radix and rehmanniae radix in a Chinese herbal formula to promote diabetic wound healing. J Ethnopharmacol 2012;141(1):250-6.

George A, Chinnappan S, Chintamaneni M, Kotak CV, Choudhary Y, Kueper T, et al. Anti-inflammatory effects of Polygonum minus (Huds) extract (Lineminusâ„¢) in in-vitro enzyme assays and carrageenan induced paw edema. BMC Complement Altern Med 2014;14:355.

Andriani Y, Effendy AW, Habsah M, Sifzizul TM. Antibacterial, radical scavenging activities and cytotoxicity properties of Phaleria macrocarpa (Scheff.) Boerl. Leaves in HepG2 cell lines. Int J Pharm Sci Res 2011;2(7):1700-6.

Hammond RA, Hannon R, Frean SP, Armstrong SJ, Flower RJ, Bryant CE. Endotoxin induction of nitric oxide synthase and cyclooxygenase-2 in equine alveolar macrophages. Am J Vet Res 1999;60(4):426-31.

Radzi SA, Andriani Y, Mohamad H, Muhammad TS, Saidin J. In vitro anti -inflammatory activities of extracts from bacteria associated with marine sponges: Theonella sp. J Teknol 2015;77(25):165-9.

Yoon WJ, Ham YM, Kim SS, Yoo BS, Moon JY, Baik JS, et al. Suppression of pro-inflammatory cytokines, iNOS, and COX-2 expression by brown algae Sargassum micracanthum in RAW 264.7 macrophages. EurAsian J Biol Sci 2009;3:130-43.

Perdicaris S, Vlachogianni T, Valavanidis A. Bioactive natural substances from marine sponges: New developments and prospects for future pharmaceuticals. Nat Prod Chem Res 2013;1(3):1-8.

Senthilkumar K, Kim SK. Marine invertebrate natural products for anti-inflammatory and chronic diseases. Evid Based Complement Alternat Med 2013;2013:572859.

Milugo TK, Omosa LK, Ochanda JO, Owuor BO, Wamunyokoli FA, Oyugi JO, et al. Antagonistic effect of alkaloids and saponins on bioactivity in the quinine tree (Rauvolfia caffra sond.): Further evidence to support biotechnology in traditional medicinal plants. BMC Complement Altern Med 2013;13:285.

Yedgar S, Lichtenberg D, Schnitzer E. Inhibition of phospholipase A(2) as a therapeutic target. Biochim Biophys Acta 2000;1488(1-2):182-7.

Penesyan A, Tebben J, Lee M, Thomas T, Kjelleberg S, Harder T, et al. Identification of the antibacterial compound produced by the marine epiphytic bacterium Pseudovibrio sp. D323 and related sponge-associated bacteria. Mar Drugs 2011;9(8):1391-402.

Kurian NK, Nair HP, Bhat SG. Evaluation of anti-inflammatory property of melanin from marine Bacillus spp. BTCZ31. Asian J Pharm Clin Res 2015;8(3):251-5.

Bitzer J, Grosse T, Wang L, Lang S, Beil W, Zeeck A. New aminophenoxazinones from a marine Halomonas sp.: Fermentation, structure elucidation, and biological activity. J Antibiot (Tokyo) 2006;59(2):86-92.

Li D, Xu Y, Shao CL, Yang RY, Zheng CJ, Chen YY, et al. Antibacterial bisabolane-type sesquiterpenoids from the sponge-derived fungus Aspergillus sp. Mar Drugs 2012;10(1):234-41.

Issa AY, Suresh VR, Wargovich MJ. The role of phytochemicals in inhibition of cancer and inflammation. New directions and perspectives. J Food Compost Anal 2006;19:405-19.



How to Cite

Andriani, Y., L. Marlina, H. Mohamad, H. Amir, S. A. M. Radzi, and J. Saidin. “ANTI-INFLAMMATORY ACTIVITY OF BACTERIA ASSOCIATED WITH MARINE SPONGE (HALICLONA AMBOINENSIS) VIA REDUCTING NO PRODUCTION AND INHIBITING CYCLOOXYGENASE-1, CYCLOOXYGENASE-2, AND SECRETORY PHOSPHOLIPASE A2 ACTIVITIES”. Asian Journal of Pharmaceutical and Clinical Research, vol. 10, no. 11, Nov. 2017, pp. 95-100, doi:10.22159/ajpcr.2017.v10i11.20094.



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