{"id":15803,"date":"2017-09-25T11:50:31","date_gmt":"2017-09-25T11:50:31","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=15803"},"modified":"2020-04-24T11:07:27","modified_gmt":"2020-04-24T11:07:27","slug":"antimalarial-effect-of-flamboyant-delonix-regia-bark-and-papaya-carica-papaya-l-leaf-ethanolic-extracts-against-plasmodium-berghei-in-mice","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol10no3\/antimalarial-effect-of-flamboyant-delonix-regia-bark-and-papaya-carica-papaya-l-leaf-ethanolic-extracts-against-plasmodium-berghei-in-mice\/","title":{"rendered":"Antimalarial Effect of Flamboyant (Delonix Regia) Bark and Papaya (Carica papaya L.) Leaf Ethanolic Extracts Against Plasmodium Berghei in Mice"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

Malaria represents a major health issue worldwide, including Eastern Indonesia. There are 433,326 clinical cases out of 232 million Indonesian population in 2005 1-3<\/sup> and malaria outbreak occurred in some areas causing 87 deaths from 18,812 population in 62 villages in Indonesia in 2005 4-7<\/sup>. The main drive for this deadly infectious disease was the development of multi-resistance of parasite populations, Plasmodium sp.<\/em> against antimalaria aminoquinolines, including chloroquine, amodiaquine and mefloquine. Plasmodium i<\/em>s a parasite that causes malaria. The typical symptoms of malaria are easily Plasmodium<\/em> is transmitted from one person to another through the intermediary of the Anopheles<\/em> mosquito.\u00a0 Some efforts to eradicate malaria are still developed so far, such as making a combination therapies of malaria drugs as a worldwide replacement most Plasmodium falciparum<\/em> isolates in endemic areas which have found multi-resistance to the aminoquinolines. Artemisinin derivatives became promising drug alternatives in combination with aminoquinolines and other drugs to treat the infection caused by multidrug resistant strains 8<\/sup>, <\/sup>9<\/sup>. However, artemisinin has been found no longer powerful to eradicate malaria due to its resistance has been reported 10-13<\/sup>, making the development of new antimalarial drugs obtained from natural sources are important.<\/p>\n

Populations in Indonesia are common in using traditional medicine due to the cultural acceptability of healers, the relatively low cost of traditional medicine and intention of less toxic remedies. The studies conducted on the traditional medicinal plants in Indonesia are extensive developed to obtain more potent medicinal plants for cancers and infectious diseases. Although recently there are efforts to identify and screen antimalarial herbs used in the ethnomedicine practice of the country, the studies done are very limited and not fully explored. Thus, pharmacological screening of medicinal plants as antimalaria is highly needed. Medicinal plants have been focused for the search of new antimalaria drugs in various parts of the world and the present global situation indicates a recent rising from malaria pecularity, due to the resistance of malaria parasites to pledge antimalaria drugs 14<\/sup>, <\/sup>15<\/sup>. Thus, it is needed to broaden research in the development of new, cheap and effective antimalaria drugs from medicinal plants. Indonesia is a tropical country which is rich of biodiversities of natural medicinal compounds to heal illnesses.<\/p>\n

Delonix regia<\/em> is a flowering plant in the pea family found in tropical areas and its leaves are used traditionally to treat diseases in folk medicine including antimalaria, especially in the area of \u200b\u200bEast Nusa Tenggara, as by drinking water decoction of Delonix regia<\/em> leaves. Delonix regia<\/em> extract was reported to have a wide range of bioactivities. The plant has shown a promising antioxidant 16<\/sup>, antimicrobial 16<\/sup>, <\/sup>17<\/sup>, antidiabetic 18<\/sup>, and anti-inflammatory effects 19<\/sup>. Delonix regia<\/em> leaf extract was also reported became a potential therapeutic agent for cardioprotection 20<\/sup>. Various solvents such as hexane, chloroform, methanol, ethanol and water were reported to be used to extract of the bark, seeds, flowers and leaves of Delonix regia<\/em> plant and have been demonstrated to possess significant antiplasmodial activity in vivo 21<\/sup>. Moreover, methanol and acetone extracts of the flower have been also reported owned good larvicidal activities 22<\/sup>, <\/sup>23<\/sup>. Meanwhile, Carica papaya L<\/em>. extracted from leaf was reported having anti-proliverative against prostate cancer, antifungal activity, antibacteria, antidengue, anti-inflamatory and antimalaria 24-30<\/sup> Here, we report our research which was designed to evaluate the bioactivity compounds from a combination of Delonix regia<\/em> and Carica papaya L.<\/em> leaves in ethanolic extract against malaria in Plasmodium berghei<\/em>-infected mice as in vivo <\/em>animal model which might be useful as potential remedies for malarial disease.<\/p>\n

Method<\/strong><\/p>\n

Plant Preparation and Extraction<\/strong><\/p>\n

The plant material used in this study was obtained from Balitro (Tropical Plant Research), the leaves of the plant were cleaned, and shielded\u00a0from sunlight, to obtain the dry ingredients, then mashed\u00a0\u00a0with a blender to get powder form. Extraction was performed by\u00a0maceration for three days then supernatant was separated, it was added with 70% ethanol until all powder was dissolved. After 3 days\u00a0immersion with stirring periodically, the solution is then filtered and\u00a0\u00a0pulp obtained by soaking several times until the filtrate was clear. The filtrate was concentrated using\u00a0 rotary evaporator at 50O<\/sup>C with 50 rpm and dried in the oven\u00a0at 35O<\/sup>C to obtain a constant weight. \u00a0Extracts were given to mice as treatments in three doses: 4.80 mg\/g\u00a0 9.75mg\/g\u00a0 and 15.5 mg\/g, respectively.<\/p>\n

Phytochemistry Assays<\/strong><\/p>\n

Phytochemistry assays were conducted to determine secondary metabolites contained in the constrated\u00a0 extract\u00a0 of\u00a0 ethanol fraction of\u00a0 Delonix regia<\/em> and Carica papaya<\/em> L. Phytochemistry assays were carried out with the following\u00a0 procedure of Clule (1982) 31<\/sup>.The phytochemical analyses including glycoside, saponin, flavonoids, alkaloids, triterpenoid, steroids, essential oil and tannin assays.<\/p>\n

Glycoside Assay<\/strong><\/p>\n

Glycoside phytochemistry assay was carried out by evaporating of 0.1 mL of sample solution over\u00a0 a water bath, then dissolve it with 5 mL acetic acid anhydride. Add 10 drops of concentrated sulfuric acid, the blue or green product is formed indicating glycosides.<\/p>\n

Saponin Assay<\/strong><\/p>\n

Saponin screening is carried\u00a0 out by placing of 10 mL of sample solution in a test tube; then it is shaken vertically for 10 seconds and then left it to stand for 10 seconds, 1-10 cm tallfoam formation that is stable for less than 10 minutes indicated the presence of saponin. For the confirmation after the addition\u00a0 of 1 drop of 2N HCl, the foam does not disappear.<\/p>\n

Flavonoids Assay<\/strong><\/p>\n

Flavonoids screening is done\u00a0 by placed of 1 mL of test solution was evaporated to dryness, the remaining moistened with\u00a0 acetone, and then added a fine powder of\u00a0 boric acidand oxalic acid, carefullyheated over a water bath and avoid overheating. Residual obtained is mixed with 10 ml.of ether. Observed with the ultraviolet 366 nm was yellow fluorescence in solution showed flavonoids.<\/p>\n

Alkaloids Assay<\/strong><\/p>\n

About 2 ml. of sample solution\u00a0 was placed on a porcelain\u00a0 cup was\u00a0 evaporated to give of residue. The residue was added 5 mL of 2N\u00a0 HCl. The resulting solution is divided into 3 tubes. The first tube serves as a blank, was added HCl 2N, the second was added 3 drops of\u00a0 Dragendorff reagent, and the third tube was added 3 drops of\u00a0 Mayer\u00a0 reagent. Positive result of\u00a0 alkoloids characterized by the formation of an orange precipitate in the second tube and the yellow precipitate in the third tube.<\/em><\/strong><\/p>\n

Triterpenoid and Steroids Assay<\/strong><\/p>\n

The triterpenoid and Steroids assays were conducted using Liebermann Burchard reaction, 2 mL of test solution was evaporated in a pocelain cup. The residue was dissolved in 0.5 mL of\u00a0 chloroform\u00a0 and acetic acid anhydride was added. Then added 2 of concentrated sulfuric acid through the tube wall. The precence of triterpenoids \u00a0is characterized by the formation of the brownish or violet ring at the boundary of the solution, where the presence of the steroids is characterized by the formation of blue-green ring.<\/p>\n

Essential Oils Assay<\/strong><\/p>\n

About 1 mL of sample solution was pipetted and then evaporated on porclain disk to obtain a residue. Possitive result of the essential oil are characterized by a distinctive odor that is generated by the residue.<\/p>\n

Tannin Assay<\/strong><\/p>\n

Tannin phytochemistry assay was conducted\u00a0 by reacting\u00a0\u00a0 of 1 mL of the\u00a0 test solution with a solution of 10%\u00a0 of\u00a0 Iron(III) Chloride, the possitive result for tannin was shown by the formation of dark blue or greenish.<\/p>\n

Inoculation of Plasmodium<\/em><\/strong> Berghei<\/strong><\/em> in Mice<\/strong><\/p>\n

The animals used in this experiment were 2-3 months old healthy male standard Swiss-Webster mice, weighing 20-30 grams. The mice were obtained from Centre for Research and Development of the Ministry of Health of the Republic of Indonesia. The mice used for antimalarial assay was initially infected with a donor of Plasmodium berghei<\/em>. Two milliliters of infected blood from donor mice\u00a0was taken from the heart by using a syringe which already contained anticoagulant. Moreover, the blood was collected in test tubes containing\u00a03.8% of sodium citrate. Blood donors were collected and injected intraperitoneally in mice as much as 0.2 ml, then incubated for five days. After incubation, blood samples were taken from the mice tail vein in order to determine whether the mice had been infected.<\/p>\n

Parasites were calculated at 500 erythrocytes and then the results were expressed in parasite density. The formula used in the parasite density calculation based on the formula given by Center for Disease Control and Prevention (CDC):<\/p>\n

\"Formula<\/p>\n

After obtaining the parasite density in each group, the increased parasitaemia could be calculated as below:<\/p>\n

Increased parasitaemia (%) = Density H4 parasite \u2013 Parasite density H0<\/p>\n

Percentage of inhibition of parasite density was determine based on the percentage of parasitaemia on the fourth day, followed Peter’s 4-day suppressive test. The standard calculation is recommended by the WHO to assess the potential antiplasmodium agent.<\/p>\n

\"Formula<\/p>\n

Descriptions<\/strong><\/p>\n

Pn = density of parasites average on day 4 in the negative control group<\/p>\n

Pu = density of parasites average on day 4 in each test group<\/p>\n

In Vivo<\/em><\/strong> Antiplasmodium Assay<\/strong><\/p>\n

Antiplasmodium activity test was performed according to the standard method of Peter’s Test (4-Days suppressive test). The blood preparations were prepared by placing one drop of blood sample, collected from the tail vein of mice into a glass microscope object. Another object glass was mounted on the preparation and the blood was smeared into a thin layer, with an angle of 30o<\/sup>. The layer air-dried, and then fixed in methanol absolute, followed by second drying. A few drops of in Giemsa dye was added to cover the entire surface. The preparation was allowed to stand for 30 minutes. Then preparation was washed with water, and the preparation was examined under a light microscope with a magnification of 10 x 100 multiplication. Mice that had been infected with the parasite were divided into 11 groups as shown in Table 1.<\/p>\n

Table 1: In vivo<\/em> treatments of Plasmodium berghei<\/em>-infected mice with plant extracts.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Mice group category<\/strong><\/td>\nPart of plant
\nextract<\/strong><\/td>\n		 <\/p>\n

(Dose mg sample\/g mice body weight)<\/strong><\/td>\n<\/tr>\n

A1<\/td>\nDelonix regia<\/em> bark<\/td>\n4.80 mg\/20 g<\/td>\n<\/tr>\n
A2<\/td>\nDelonix regia<\/em> bark<\/td>\n9.75 mg\/20 g<\/td>\n<\/tr>\n
A3<\/td>\nDelonix regia<\/em> bark<\/td>\n15.50 mg\/20 g<\/td>\n<\/tr>\n
B1<\/td>\nCarica papaya L <\/em>leaf<\/td>\n4.80 mg\/20 g<\/td>\n<\/tr>\n
B2<\/td>\nCarica papaya L <\/em>leaf<\/td>\n9.75 mg\/20 g<\/td>\n<\/tr>\n
B3<\/td>\nCarica papaya L <\/em>leaf<\/td>\n15.50 mg\/20 g<\/td>\n<\/tr>\n
C1<\/td>\nDelonix regia<\/em> bark +\u00a0 Carica papaya L <\/em>leaf<\/td>\n1:1<\/td>\n<\/tr>\n
C2<\/td>\nDelonix regia<\/em> bark +\u00a0 Carica papaya L <\/em>leaf<\/td>\n1:3<\/td>\n<\/tr>\n
C3<\/td>\nDelonix regia<\/em> bark +\u00a0 Carica papaya L <\/em>leaf<\/td>\n3:1<\/td>\n<\/tr>\n
D<\/p>\n

(water as negative control)<\/td>\n

–<\/td>\n–<\/td>\n<\/tr>\n
E<\/p>\n

(Sulfadoxine as positive.control)<\/td>\n

–<\/td>\n1,36 mg\/20 g<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Sample size used in this research was following Federer\u2019s formula 31<\/sup>:<\/p>\n

(k-1) (n-1) > 15<\/p>\n

(11 – 1) (n – 1) > 15<\/p>\n

n ~ 3<\/p>\n

Descriptions<\/strong><\/p>\n

k = number of groups<\/p>\n

n= number of mice in groups<\/p>\n

Results<\/strong><\/p>\n

The phytochemical analyses of the ethanolic extracts of Delonix regia<\/em> and Carica papaya<\/em> L were done, qualitatively. The plants were collected and were identified. Then they were shade dried and powdered and were subjected to phytochemical screening. The qualitative chemical tests for the ethanolic extracts were performed. The investigation showed that Delonix regia<\/em> contains flavanoids, alkaloids, triterpenoid and tannins. Meanwhile, Carica papaya<\/em> L. screening showed that the ethanolic extract possesses glycosides, flavanoids, alkaloids, triterpenoid, steroids and tannins, as show on Table 1.<\/p>\n

Table 2: Phytochemistry analysis extract ethanol of\u00a0 Delonix regia<\/em> and\u00a0<\/strong>Carica papaya<\/em><\/strong> L.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
Phytochemistry name<\/strong>\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/strong><\/td>\nDelonix regia\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/em><\/strong><\/td>\nCarica papaya L.<\/em><\/strong><\/td>\n<\/tr>\n
Glycosides<\/td>\n–<\/td>\n+<\/td>\n<\/tr>\n
Saponin<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
Flavonoid<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n
Alkaloid<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n
Triterpenoid<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n
Steroid<\/td>\n–<\/td>\n+<\/td>\n<\/tr>\n
Essential oil<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
Tannin<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

The treatment effect of sulfadoxine inhibition, ethanolic Delonix regia<\/em> extract in Plasmodium berghei<\/em>-infected mice, are tabulated in Table 1. Sulfadoxine induction in ill mice, showed 100% inhibition rates. Meanwhile, 9.75 mg\/g Delonix regia<\/em> extract showed 66.25% inhibition rate, followed by 4.80 mg\/g Delonix regia<\/em> \u00a0and 15.50 mg\/g Delonix regia<\/em> extracts were 38.88% and 27.36%, respectively as depicted on Figure 1. In addition, Plasmodium berghei<\/em> \u00a0growth in mice mostly decreased by treatment of Delonix regia <\/em>with concentration 9.75 mg\/g as shown on Table 2.<\/p>\n

Table 3: The growth and inhibition rates of parasite density on the 4th<\/sup> day of\u00a0<\/strong>treatment with Delonix regia <\/em>bark extract<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/strong>Treatment Group\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/strong><\/td>\n% Growth<\/strong><\/td>\n% Inhibition<\/strong><\/td>\n<\/tr>\n
Positive control<\/td>\n0.00<\/td>\n100.00<\/td>\n<\/tr>\n
Delonix regia <\/em>(4.80 mg\/g)\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/em><\/td>\n79.34<\/td>\n38.88<\/td>\n<\/tr>\n
Delonix regia <\/em>(9.75 mg\/g)\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/em><\/td>\n42.35<\/td>\n66.25<\/td>\n<\/tr>\n
Delonix regia\u00a0 <\/em>(15.50 mg\/g)\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/em><\/td>\n80.04<\/td>\n27.36<\/td>\n<\/tr>\n
Negative control<\/td>\n100.00<\/td>\n0.00<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"Figure<\/td>\nFigure 1: Inhibition activities of positive control and Delonix regia <\/em>bark extract in various concentrations.<\/strong><\/p>\n

 <\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Table <\/strong>4<\/strong>: The growth and inhibition rates of parasite density on the 4th<\/sup> day of\u00a0<\/strong>treatment with Carica papaya<\/em> <\/strong>L <\/strong>extract<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Treatment Group<\/strong><\/td>\n% Growth<\/strong><\/td>\n% Inhibition<\/strong><\/td>\n<\/tr>\n
Positive control<\/td>\n0.00<\/td>\n100.00<\/td>\n<\/tr>\n
Carica papaya <\/em>L (4.80 mg\/g)<\/td>\n19.43<\/td>\n83.75<\/td>\n<\/tr>\n
Carica papaya <\/em>L (9.75 mg\/g)<\/td>\n29.00<\/td>\n89.02<\/td>\n<\/tr>\n
Carica papaya <\/em>L (15.50 mg\/g)<\/td>\n27.02<\/td>\n72.89<\/td>\n<\/tr>\n
Negative control<\/td>\n100.00<\/td>\n0.00<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\u00a0\"Figure<\/td>\nFigure 2: Inhibition activities of positive control and Carica papaya<\/em> L leaf extract in various concentrations.<\/strong><\/p>\n

 <\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Treatment effect result of sulfadoxine inhibition in Figure 2 showed that ethanolic Carica papaya<\/em> L extract in Plasmodium berghei<\/em>-infected mice was found 100% inhibition rate. Meanwhile, 9.75 mg\/g Carica papaya<\/em> L extract showed 89.02%, followed by 4.80 mg\/g Carica papaya<\/em> L and 15.50 mg\/g Carica papaya<\/em> L extracts were 83.75% and 72.89%, respectively as depicted on Figure 1. In addition, Plasmodium berghei<\/em> \u00a0growth in mice mostly decreased by treatment of Carica papaya<\/em> L with concentration 9.75 mg\/g as shown on Table 3.<\/p>\n

Table <\/strong>5<\/strong>:<\/strong> The inhibition rates <\/strong>of <\/strong>parasite density on the 4th<\/sup> day of combination treatment with <\/strong>Delonix regia <\/em><\/strong>and Carica papaya<\/em><\/strong> L <\/strong>extract<\/strong>s<\/strong><\/p>\n\n\n\n\n\n\n\n\n
Treatment Group<\/strong><\/td>\n% Inhibition<\/strong><\/td>\n<\/tr>\n
Positive control<\/td>\n100.00<\/td>\n<\/tr>\n
Combination 1:1<\/td>\n97.00<\/td>\n<\/tr>\n
Combination\u00a0 3:1<\/td>\n29.03<\/td>\n<\/tr>\n
Combination\u00a0 1:3<\/td>\n6.01<\/td>\n<\/tr>\n
Negative control<\/td>\n0.00<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\u00a0\"Figure<\/td>\nFigure 3: Inhibition activities of positive control and a combination of Delonix regia <\/em>bark with Carica papaya<\/em> L leaf extracts in various ratio concentrations: 1:1, 3:1 and 1:3, respectively.<\/strong><\/p>\n

 <\/p>\n

 <\/p>\n

Click here to View figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Combination treatment of ethanolic Delonix regia<\/em> and Carica papaya<\/em> L extracts was also investigated its inhibition rates of parasite density on the 4th<\/sup> day as showed on Table 4. It showed that combination extracts in ratio 1:1 was found 97% the highest inhibition Plasmodium berghei<\/em> in mice. \u00a0Followed by Delonix regia <\/em>bark with Carica papaya<\/em> L leaf extracts in ratio concentrations 3:1 and 1:3 were 29.03% and 6.01%, respectively. It indicated that combination extracts in ratio 1:1 was potential bioactivities as well as sulfadoxine inhibition as positive control (Fig.3).<\/p>\n

Discussion<\/strong><\/p>\n

The study of the bioactive compounds of the medicinal plants have acquired a lot of importance all over the world. The present study includes the phytochemical screening of the plants Delonix regia<\/em> and Carica papaya<\/em> L. The qualitative analyses of the ethanolic extracts from Delonix regia<\/em> bark and Carica papaya<\/em> L leaf showed the presence of phytochemical constituents, including flavonoid, alkaloid, triterpenoid, steroid, tannin and glycosides. These compounds are potentially used for various treatments as herbal remedies. Thus, the development of medicinal plants will be very useful for combating diseases 26, 33, 34<\/sup>.<\/p>\n

The results of the study confirms the effectiveness of\u00a0 the use of extract plants is the easiest, reliable and the most practicable method, in inhibiting parasite density in Plasmodium<\/em> berghei<\/em>-infected mice.\u00a0 In this study, ethanolic Delonix regia<\/em> bark extract potentially reduced parasite density down to 42.35% (0% growth rate with positive control), while in the previous study reported that bark and fruit peel extract of Delonix regia<\/em> (72.8 mg\/kg ) was found to be highly effective producing inhibition of 122% and 117% with hexane, chloroform, methanol, ethanol and water solvent extractions 33<\/sup>.<\/p>\n

Meanwhile, ethanolic Carica papaya<\/em> L leaf extracts potentially reduced parasite density down to 29% (0% growth rate with positive control). It is showed that Carica papaya<\/em> L leaf extracts has higher inhibition activity compare to Delonix regia<\/em> bark extract on the same single dose. The previous study reported that Carica papaya<\/em> fruit and leaf extracts were potential antilarvicidal and antimalaria in vitro and in vivo 26<\/sup>, <\/sup>33<\/sup>, 34<\/sup>.<\/p>\n

Based on extract plants examination, it can be seen that the positive control has the highest \u00a0inhibition rate. In clinical doses, the combination of Delonix regia<\/em> and Carica papaya <\/em>L. At 1:1 ratio has the highest antiplasmodium activity compared to other concentration ratios observed in this research. From previous studies it has been demonstrated that the bark of Delonix regia<\/em> have antimalarial effects in mice infected with Plasmodium<\/em> berghei<\/em>.\u00a0 Phytochemical studies showed that Delonix <\/em>bark extract contained natural compounds such as alkaloids, flavonoids, terpenoids, and phenolics 3<\/sup>3<\/sup>.<\/p>\n

A combination of two or more drugs or plant extracts can produce a synergistic or antagonist effects to Plasmodium<\/em> berghei<\/em> in mice. The synergistic effect occurs when the combination mutually enhance the efficacy of each ingredient. Antagonistic effect occurs when an ingredient in a combination reduces the activity of another ingredient, resulting in lower efficacy then in individual dose. If drugs or plant extracts combined do not interact, but only lead to accumulated effect, it is said that the combination has an additive effect. Thus, combinations of ethanolic extracts of Delonix regia<\/em> and Carica papaya <\/em>L \u00a0in ratio 1:1 was giving the optimum anti-plasmodial activity (synergistic or additive effects) in vivo<\/em>, which were suggested to be the effective medicinal plants antimalarial potential in mice.<\/p>\n

Conclusion<\/strong><\/p>\n

In conclusion, thia Study has demonstrated potential anti-plasmodial activity for the treatment of malaria. The enhanced efficacy of certain combinations of herbal extracts remotely explains the practice and use of combined medicinal plant extracts in the management of malaria by traditional health users. This study revealed that combination of \u00a0Delonix regia<\/em> and Carica papaya <\/em>L \u00a0had much better anti-plasmodial properties compared to each single dose plant extract, and it should be subjected to further chemical and biochemical investigations. Moreover, natural products from plants used in traditional medicine, which have potent anti-plasmodial activity represent potential sources of new anti-malarial remedies. It is conceived further studies on these plants may lead to development of new affordable and effective phytomedicines for malaria.<\/p>\n

Acknowledgement<\/strong><\/p>\n

We would like to thank Mr. Winarno and staff in Center for Research and Development of the Ministry of Health of the Republic of Indonesia for financially supported this study.<\/p>\n

Conflict of Interest<\/strong><\/p>\n

Authors declare that there is no conflict of interest.<\/p>\n

References<\/strong><\/p>\n

    \n
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