EFFECTIVENESS OF ANTIBACTERIAL EXTRACT OF KENOP (GOMPHRENA GLOBOSA) FLOWER EXTRACT AGAINST GROWTH OF PROPIONIBACTERIUM ACNES BACTERIA

Article History Submitted:01/05/2020 Accepted:02/082020 Published:01/09/2020


INTRODUCTION
Acne (Acne vulgaris) is a chronic inflammation of the pilosebaceous tissue caused by Propionibacterium acnes (P.acnes) bacteria which is characterized by the appearance of blackheads, pustules, lesions, papules, and nodules measuring 1-5 mm [1]. The incidence for acne were 26% in women, 12% of men with aged 40 years old, and 20% of adolescents [2]. Base on the literature, approximately 81% people with acne are influenced by genetic factors, and the rest is influenced by external factors such as air pollution, race, excessive fatty food, etc. It also easily appears on oily facial skin and during premenstruation on for women due to hormonal influences. Acne often affects areas of the face so that it interferes with daily activities, cause lack of confidences and 30% -50% of psychiatric disorders [3].
The pathogenesis of rupture, consists of follicular epidermal hyperproliferation, excessive sebum production, and the appearance of P.acnes bacteria. Propionibacterium acnes produces extracellular lipase, which replaces inflammation in the skin and overcomes acne [4]. The wall of the P.acnes bacteria consists of antigens that stimulate antibody formation. Anti-Propionibacterium anti-bodies increase the proinflammation cascade. This bacteria stimulates cytokine binding that binds to Toll-Like Receptor 2 (TLR-2) on monocytes and poly-morphonuclear cells in the sebaceous follicle region, instead, using pro-inflammatory cytokines such as IL-1α, IL-8, IL-12, and TNF -α [3]- [5].
Pharmacotherapy for P.acnes bacteria is often used is benzoyl peroxide, antibiotics, or retinoids, but retinoid often cause side effects such as erythema, itching, rashes, even pseudomembranous colitis and also experience resistance such as metronidazole and phosphomimic groups [6]. Clindamycin drug could be using against P.acnes. In addition, clindamycin was use as positive control that effective to inhibit the growth of P.acnes bacteria.
There are 30,000 species of medicinal plants in Indonesia, and Indonesian citizens use only 1,200 species [7]. One of the medicinal plants that are often used is the kenop flower (Gomphrena globosa). The Kenop flower has a length of 10-15 cm and is commonly used as ornamental plants and in Bali traditional ceremonies. Kenop flower (Gomphrena globosa) in the basic contains such as carbohydrates, antifungal, glycosides, phenols, koumarins, steroids, anthraquinones compounds, color pigments betacyanin, anti-cancer, analgesics, gomphrenin III which are antiinflammatory and antioxidant substances [7], [8]. According to Kusmiati research in 2017, Kenop flower extract (Gomphrena globosa) contains antibacterial substances that can inhibit the growth of the Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, and Salmonella typhi bacteria [8]. Antioxidants in Gomphrena globosa flowers such as tannins, flavonoids, saponins, quinones, and alkaloids block the mechanism of metabolism and interfere with the permeability of bacterial cells so that they inhibit bacterial growth. Furthermore, the research for effectiveness from Kenop flower against P.acnes bacteria not already yet. Therefore, we conducted a study to find out the effect of kenop extract on the inhibitory growth of P.acnes bacteria.

RESEARCH METHOD
This research is an experimental study carried out at the Microbiology Laboratory of Udayana University from July to September 2019. The study has received an ethical eligibility permit from the Research Ethics Commission (KEP) of the Faculty of Medicine, Udayana University, with letter number 1825 / UN14.2.2.VII.14 / LP / 2019. The independent variable in the study was the methanol extract of kenop flower (Gomphrena globosa), which was tested on P.acnes bacteria. The dependent variable is the diameter of the inhibitory zone in P.acnes bacteria. Controlled variables were temperature and incubation time, that was repeated three times to ensure validity. There were five samples in one experiment, the concentration for samples are from Kenop flower extract such as, 25%, 50%, 75%, 100%, and also clindamycin for positive control.
Primary data obtained from the Kirby-Bauer experimental method with Muller-Hinton media. Secondary data was obtained from literature studies, journals, literature reviews, textbooks, and the internet. The steps in this study, firstly, was to manufacture a simplicial and methanol extract using the kenop (Gomphrena globosa) flower. Following that, was to make the bacterial culture using the Kirby-Bauer method, and lastly, would be to test for antibacterial activity, and to measure the data using the SPSS software application version 16.00. From 100 grams kenop (Gomphrena globosa) flower simplicial, around 2 grams of a thick extract of the kenop (Gomphrena globosa) was obtained.The test solution consists of 4 groups, namely group 1 methanol extract concentration of 100% methanol kenop (Gomphrena globosa) flower, group 2 with 75% concentration, group 3 with 50% concentration, and the last group concentration was 25%. The Positive control solution used is clindamycin phosphate 1.2%, and the negative control used is distilled water. Propionibacterium acnes bacteria was isolated on blood agar media and incubated at 37ºC for 24 hours, and its form and nature have been proven by gram staining.

RESULTS AND ANALYSIS
The results of the measurement of the inhibition of these bacteria are measured using calipers in millimeter (mm) ( Table 1 and Figure 1).  The normality test uses Shapirowilk and shows that the data is normally distributed so the average difference test is using One Way ANOVA because the average difference is more than two groups (Table 1). Based on Table 2, it can be concluded that the higher the extract, the greater the inhibitory power, as long as the extract has not reached saturated concentration (CI = 95% and p <0.05). The lowest extract concentration needed to cause inhibition is 25%. The average inhibition on control + 15.6 mm, control -0 mm, concentration of 100% 7.25 mm, concentration 75% 11.33 mm, concentration 50% 9.33 mm and concen-tration 25% 6.5 mm.
The inhibitory zone from concentration of 75% is classified as a strong inhibitory zone while the inhibitory zones of 100%, 50%, and 25% are classified as moderate. This is consistent with research conducted by Manaf and colleagues in 2013, which stated that the provisions of the antibacterial strength of a plant are inhibitory zones. [9] If the inhibition zone is equal to 20 mm, the inhibition zone is sensitive, the inhibition zone of 10-20 mm is classified as sensitive, 5-10 mm is classified as moderate, and the inhibition zone below 5 mm is classified as resistance [9], [10]. According to Hidayah's statement in 2016, a microbe is declared sensitive to antimicrobials if the anti-microbial substances in plant extracts have a inhibition zone of 12-24 mm [10]. The results show that only kenop (Gomphrena globosa) flower extract with a concentration of 75% are sensitive to P.acnes bacteria with minimum inhibitory zone 11.3 mm which is close to a minimum standard of antimicrobials that are sensitive to bacteria. Clindamycin antibiotic as the positive control is bacteriostatic and bactericidal and also has a narrow spectrum antibiotic group because it only works on gram-positive bacteria. Its performance is influenced by the concentration of the drug, the causative organism and the area of infection of the organism [6], [10]- [12].
In general, the increase in diameter of the inhibitory zone of the extract in bacterial activity increases with the concentration of the extract being concentrated. However, this study found a decrease in inhibition zone diameter at a concentration of 100% due to differences in the diffusion rate of antibacterial compounds on agar media, types and concentrations of antibacterial compounds, so as to give a difference in zone diameters within a certain period of time [6], [10], [13], [14].
The phenol component in flavonoids denaturizes enzymes and amino acids and activates essential enzymes in P. acnes bacteria [6], [11]. It caused bacterial cells nutrient leakage due to damage to cells membrane hydrophobic bonds such as protein and phospholipid bonds thus inhibiting the bacterial replication [10].
Tannin compounds bind to the building blocks of H + bacterial protein walls.That could effect the PH becomes acidic and makes the protein denature, thereby inhibiting bacterial development. Tannins also inhibit DNA topoisomerase and reverse transcriptase enzymes so that bacterial cells cannot replicate [8], [11], [15], [16].
Alkaloid compounds interfere with the peptidoglycan constituent in bacterial cells which cause the cell wall layer not to be formed as a whole due to a disruption in the process of active transport function, selectively the permeability function and protein composition, so that it will cause the cell death slowly while saponin compounds work by inhibiting and even killing bacteria by decreasing cell surface tension, causing the bacteria to undergolysis [8], [9], [14], [16].

CONCLUSION
Kenop flower (Gomphrena globosa) extract has good bacterial inhibition, especially from concentration of 75% with a sensitive inhibitory zone category. The conclusion of this study is Knobs flowers has a great potential anti-bacterial against P.acnes bacterial. Further research needs to be done on the toxicity and bactericidal effects of kenop (Gomphrena globosa) flower extracts on other microorganisms. One of the potential results that can be done, is by making the extract of the kenop (Gomphrena globosa) flower into an acne herbal medicine which will continue to be available in phytopharmacology capsulation after several tests in subsequent studies.