Vol 12, Issue 12, 2019 Online - 2455-3891 Print - 0974-2441 ANTI-INFLAMMATORY ACTIVITY OF MARINE SPONGE CALLYSPONGIA SP. AND ITS ACUTE TOXICITY ADRYAN FRISTIOHADY 1 , WAHYUNI WAHYUNI 1 , FADHLIYAH MALIK 1 , LA ODE MUHAMMAD JULIAN PURNAMA 1 , BARU SADARUN 2 , SAHIDIN I 1 * 1 Department of Pharmacy, Faculty of Pharmacy, Universitas Halu Oleo, Kendari 93232, Southeast Sulawesi, Indonesia. 2 Department of Fisheries, Faculty of Fisheries and Marine Science, Universitas Halu Oleo, Kendari 93232, Southeast Sulawesi, Indonesia. Email: sahidin02@uho.ac.id Received: 30 June 2019, Revised and Accepted: 23 October 2019 ABSTRACT Objective: This study aims to investigate the anti-inflammatory effect of the ethanolic extract of Callyspongia sp. using stabilization of the human red blood cell (HRBC) membrane method and its acute toxicity using brine shrimp lethality test (BSLT) method. Methods: Callyspongia sp. was macerated with 96% ethanol. Extract characterized and screened for the secondary metabolite. Anti-inflammatory activity by stabilization of the HRBC membrane method with a varied dose of 50 ppm; 100 ppm; 200 ppm; 400 ppm; 800 ppm; 1600 ppm; and 3200 ppm. Solutions observed using a photometer to describing stability and ability in preventing membranes hemolytic and statistically analyzed using SPSS. Acute toxicity carried out by the BSLT method and analyzed using Minitab ® ver. 17.2.1. Results: The phytochemical screening was indicating that Callyspongia sp. contains flavonoid, alkaloid, and terpenoid. The results of the anti- inflammatory activity test showed that the percentage value of stability and hemolysis of extracts with doses of 50, 100, 200, 400, 800, 1600, and 3200 ppm were 55% and 45%, 63% and 37%, 70% and 30%, 74% and 26%, 80% and 20%, 87% and 13%, and 97% and 3%, respectively. It showed that extract of sponge Callyspongia sp. in all varied dose has activity in stabilizing the HRBC membrane thus can be potential as an anti-inflammatory. The results of acute toxicity assay showed that the value of LC 50 was 1281.45 μg/ml and categorized as nontoxic to Artemia salina Leach. Conclusion: Various concentrations of Callyspongia sp. effective as an anti-inflammatory in stabilizing HRBC, and categorized as safe. Keywords: Anti-inflammatory, Callyspongia sp., Red blood cell membranes. INTRODUCTION Inflammation is a response of host against to antigen which can cause tissue injury such as infection and burns hence the antigen will not spread. Inflammation is characterized by cardinal signs which are rubor, tumor, calor, dolor, and functiolesia. Inflammation is an important in the healing process however it can disturb activities if suffered for long term. Thus, the anti-inflammation agent is needed to decrease and stop the inflammation process [1]. Callyspongia sp. is one of marine sponges which can be found in Indonesia’s abundant sea. Callyspongia sp. has activity as antibacterial, antifungal, promoting antitumor, antiretroviral, and anti-inflammation [2,3]. Previous studies showed that Callyspongia sp. contains triterpenoid, steroid, alkaloid, and flavonoid [4-6]. Flavonoid and terpenoid contained in Callyspongia sp. potentially have anti- inflammatory activity by its ability to stabilize the membranes. Human red blood cell (HRBC) membranes can be used as a parameter for in vitro anti-inflammatory activity due to its similarity with lysosome membranes, which are responsible for the inflammatory process [1,7]. Callyspongia sp. can be developed as a new alternative in treating inflammation because commercial anti-inflammation drugs such as diclofenac sodium if consumed for long term has side effects. However, each study that involved natural products that have potency as drug or empirical used as drug, need pre-clinical toxicity acute assay to predict the safety following other pharmacological tests [8]. Many previous studies focus in researching marine sponges in each aspect, yet remains have no study reported about anti-inflammatory properties of Callyspongia sp. using stabilization of the HRBC membrane method. Therefore, this study aims to investigate the anti-inflammatory activity of the ethanolic extract of Callyspongia sp. with stabilization of the HRBC membrane method and its acute toxicity acute to Artemia salina Leach larvae. METHODS Marine sponge extract Marine sponge of Callyspongia sp. was determined at Faculty of Fisheries and Aquaculture, Universitas Halu Oleo, Kendari (No. 0087/ UN29.12.1.UPP/2018). Marine sponge obtained from Bintang Samudra Edu-Marine Park, Konawe Regency, Southeast Sulawesi with a total of 2.8 kg. Marine sponge then wet sorted and chopped into pieces. The pieces were macerated with 96% ethanol for 3×24 h. Filtrated obtained was concentrated using rotary vacuum evaporator (Rotavapor, Buchi ® ) at temperature 60°C and water bath (60°C) yielded concentrated extract 31.52 g (1.12%). Characterization of extract Characterization of extract conducted was moisture content and ash content. Moisture content 2 g of extract was put in the oven (105°C) for 3–5 h then, cooled in desiccators for 30 min. Extract then weighed until constant. © 2019 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ajpcr.2019.v12i12.34737 Research Article