Level of Cytokine Interleukin-6 and Interleukin 1-β on Infectious Rat Model Treated with Etlingera elatior (Jack) R.M. Smith Fruit Extract as Immunomodulator

Etlingera elatior (Jack) R.M Smith or locally in Southeast Sulawesi known as wualae fruit has activity as an immunomodulator by increasing phagocytosis activity. Prior studies have been conducted to observe the effect of E. elatior as an immunomodulator, thus further study is needed to observe the production of cytokines such as IL-1β and IL-6 which are responsible for the immune responses. Etlingera elatior fruit macerated with 96% ethanol for three days and produced a total of ±74.6 g concentrated extract. Experimental animals used were divided into four groups (n=4) and treated orally once per day for seven days as follows: group I (0.5% Na-CMC); group II (Stimuno®); group III (E. elatior fruit extract concentration of 300 mg/kg BW); and group IV (E. elatior fruit extract concentration of 400 mg/kg BW). On the eight-day, animals were infected with Staphylococcus aureus intraperitoneally and left for an hour. Thereafter, blood was collected and assayed using ELISA Kit (Elabscience rat IL-1β and Elabscience rat IL-6). Results demonstrated that group IV increased levels of IL-1β and group III and IV increased level of IL-6 (p<0.05). Increased levels of IL-1β and IL-6 are associated with phagocytosis in the immune response. In conclusion, E. elatior fruit extract at concentration of 300 and 400 mg/kg BW increases levels of IL-1β and IL-6.


INTRODUCTION
Etlingera elatior (Jack) R.M. Smith is one of the spices native to Indonesia which belongs the Zingiberaceae family. It has been used traditionally used as medicines and flavor enhancers (Farida & Maruzy, 2016;Syarif et al., 2015). Etlingera elatior fruit is known as wualae by people in the Konawe region of Southeast Sulawesi as spices.
Besides that, it empirically used as traditional medicine as immunomodulator in recovery of typhoid fever in the

Level of Cytokine Interleukin-6 and Interleukin 1-β on Infectious Rat Model Treated with Etlingera elatior (Jack) R.M. Smith Fruit Extract as Immunomodulator
North Kolaka district of Southeast Sulawesi Wahyuni et al., 2017). Immunomodulator is drugs that modify response immune system increasing the production of serum antibodies, namely immunostimulators, thus enhance immune responses against infectious diseases, tumours and immunodeficiency (primary or secondary); and by decreasing production of serum antibodies, namely immunosuppressive, thus reduce the immune responses against autoimmune disease or transplanted organs (Catanzaro et al., 2018;Bascones-Martinez et al., 2014).
Previous studies reported that E. elatior methanol extract contains flavonoids, phenolics, alkaloids, and saponins.
Flavonoids can increase the activity of lymphocyte proliferation thus prevents invasion of viruses, bacteria, and other microbes (Panche et al., 2016). Further studies demonstrated that phagocytic activity of macrophages increase after administration of E. elatior Wahyuni et al., 2017). Increased activation of macrophage and lymphocytes stimulate secretion of cytokines such as interleukin (IL)-1β and IL-6 (Kany et al., 2019). They play important role in inflammatory reactions and induces antibody production (Besung et al., 2016;Duque & Descoteaux, 2014  Animal blood was then collected with cardiac puncture as much as 3 mL and put in a test tube containing EDTA.

Material
The tube was then centrifuged for 15 minutes at 3000 rpm and a temperature of 25°C. Blood plasma was then tested using the ELISA Kit (Elabscience® rat IL-1β and Elabscience® rat IL-6).

RESULTS AND DISCUSSION
Administration of extracts in experimental animals is carried out for seven consecutive days orally once per day, which aims to stimulate the immune system of each group of experimental animals. On the eighth day, S.
aureus inocula is injected into animals intra peritoneally.
Staphylococcus aureus is gram-positive bacteria that can cause infections both in humans and animals. They do not produce protein A, which is an antiphagocyte protein, causing S. aureus unable to avoid phagocytosis of peritoneal macrophages (Hariyanti et al., 2015;Wahyuni et al., 2017). After being injected with a bacterial inocula, all the experimental groups were left for an hour to make the innate immune system work. The innate immune system can be active within the range of 0-12 hours after infection (Abbas et al., 2016).
Macrophages and neutrophils, including the first line of defense in the immune system. Macrophages are able to fight off infections for about an hour before the immune mechanism is mobilized. On this basis, macrophage taking is done one hour after bacterial induction, so it can be seen the extent of the ability of macrophages to conquer bacterial invasion (Chaplin, 2010). Macrophages enter the site of infection are increased and the phagocytic ability of antigens also increases. In addition, levels of IL-1β and IL-6 also increased (Besung et al., 2016).