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iC*ME

THE 1ST INTERNATIONAL CONFERENCE ON HYPERBARIC, UNDERWATER AND COASTAL MEDICINE FACULTY OF MEDICINE, HANG TUAN UNIVERSITY, SURABAYA, INDONESIA

CURRENT RESEARCH

ON HYPERBARIC, UNDERWATER AND COASTAL MEDICINE

Theme:

Hyperbaric and Underwater Medicine for the Future

needing noo/ø

27-28 July 2018 Four Points Hotel Surabaya, Indonesia

Publisher:

Fakuttas Kedokteran Universitas Hang Tuah

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THE INTERNATIONAL CONFERENCE ON HYPERBARIC, UNDERWATER AND COASTAL MEDICINE (ICOME) FACULTY OF MEDICINE, HANG TUAH UNIVERSITY, SURABAYA, INDONESIA

Current Research on Hyperbaric, Underwater and Coastal Medicine

Theme:

Hyperbaric and Underwater Medicine for the Future

needing Zoo/e

27-28 July 2018 Four Points Hotel Surabaya, Indonesia

FAKULTAS KEDOKTERAN UNIVERSITAS HANG TUAH

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The 11t International Conference on Hyperbaric, Underwater and Coastal Medicine (ICOME) Faculty of Medicine, Hang Tuah University, Surabaya, Indonesia

Current Research on Hyperbaric, Underwater and Coastal Medicine Theme: Hyperbaric and Underwater Medicine for the Future

Organizing Committee:

Chairman Co-chair

Secretariat division

Registration

Proceeding Book

Janto Poemomo Hadi, dr., SpP.,SpKL Choesnan Effendi, dr., AIF.,AIFO Wienta Diarsvitri, dr., M.Sc., PhD Dr. Erina Yatmasari, dr., M.Kes Prima Arundani, dr.

Mita Herdiyantini, dr.,SpOG Treasurer

Steering Committee:

Suwarno,

Dr.lr. Sudirman, s.1P., SE., M.AP., M.H Dr. Hj. Dian Mulawarmanti, drg., M.S.

Hadi Soesilo, dr, Sp.M

Ir. Sudyantoro Hadi, M.Si (Han) Sakti Hoetama, dr, Sp.U

Reviewer:

Dr. Erina Yatmasari, dr, M.Kes Dr. Titut Hernanik, dr, M.Kes Dr. Fitri Handajani, dr, M.Kes Dr. Herin Setyaningsih, dr, M.Kes Dr. Prawesty Diah Utami, dr, M.Ked Editorial Board:

Prof. Michael Heywood Bennett Prof. Angel Anne Yanagihara Dr. Retno Budiarti, dr, M.Kes Dr. Sulistiana Prabowo, dr, MS Editor:

Wienta Diarsvitri, dr, M.Sc, Ph.D

Cover designer, setting/layout:

Wienta Diarsvitri, dr, M.Sc, Ph.D Publisher :

Fakultas Kedokteran Universitas Hang Tuah

Jalan Gadung No. I (Kompleks Barat RSPAL Dr. Ramelan) Surabaya 60244.

Telp. 031-8438750, Fax. 031-8433626 First publication in December 2019.

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The Effect of Curcuma (Curcuma xanthorrhiza Roxb.) Rhizome Extract to

^the

Amount of Leukocytes and Haemoglobin in Male BALB/c Mice

(Mus musculus L.) Infected by Plasmodium berghei ANKA

Prawesty D. Utamil*, Taufan Wiryakusuma2, Nugroho Y. Abriyant03, Anindya K. Winahyu4•Azarine Neira IDepartement ofParasitology, Faculty of Medicine, Hang Tuah University, Surabaya, Indonesia

2J•4SFaculty ofMedicine, Hang Tuah University, Surabaya, Indonesia

*Corresponding Author: [email protected]

Abstract

Malaria causes rupture of erythrocytes, thereby decreasing the amount of hemoglobin. The effect of ^malaria on the picture of leukocyte count remains unclear. Malaria disease management is also limited due to resistance. Curcuma (Curcuma xanthorrhiza Roxb.) which is known to contain curcumin and flavonoids that have antiplasmodium, antiinflammation and antioxidant effects. This research want to preceive the effect of curcuma extract on leukocyte and haemoglobin level on male BALB/C mice (Mus musculus L.) infected by Plasmodium berghei ANKA. This research was conducted experimentally post-test only control group on five groups of mice. One group was nonnal while four other groups were inoculated with Plasmodium berghei ANKA. One out ouf4 inoculated group was given aquadest and three groups were treated with curcuma extract at 150 mg/KgBW, 100 mg/KgBW, and 50 mg/KgBW dose for four days. On the fifth day, a blood test was performed to determine the number of leukocytes and haemoglobin. The statistical test showed p = (significant) which proved hypothesis that giving curcuma extract influenced the number of leukocytes and hemoglobin on male BALB / c inoculated with Plasmodium berghei ANKA. Group with extract dose 150 mg/KgBW showed the most effective decrease in parasitemia levels and significantly lower leukocyte and haemoglobin levels. Curcuma extract (Curcuma xanthorrhiza Roxb.) may affect the number of leukocytes and haemoglobin ofmale BALB/C mice (Mus musculus L.) infected with Plasmodium berghei ANKA.

Keywords: Malaria, curcuma (Curcuma xanthorrhiza Roxb.), Leukocyte, haemoglobin, Plasmodium berghei

INTRODUCTION

Malaria is a global health problem especially in tropical and subtropical countries, which is caused by the Plasmodium parasite and transmitted by Anopheles mosquito(Paniker & Ghosh 2013;

UNICEF & World Health Organization 2015).

Laboratory findings of malaria infection that are often found are changes in blood parameters

especially leukocytes and erythrocytes. The pathophysiologic mechanism for malaria induced anaemia involve destruction of infected erythrocyte, liberation of parasite and erythrocyte materials into circulation and the host reactions to these events (Ekaidem & Akpan 2016). Severe

anemia and tissues damage may occur in the phase of hyperparasitemia or excessive parasite growth (Wahlgren & Perlmann 2005; The Ministry of Health Republic of Indonesia 2015; UNICEF &

World Health Organization 2015).White blood cell

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(WBC) counts during malaria are generally characterized as being low to normal, a phenomenon that is widely thought to reflect localization of leukocytes away from the peripheral circulation and to the spleen and other marginal pools, rather than actual depletion or stasis, Leukocytosis is typically may be associated with concurrent infections and/or poor prognosis (McKenzie et al. 2005; Gansane et al. 2013). Some studies illustrate that malaria toxin glycosyl phosphatidylinositol play directly on immunological response by activation monocytes and macrophages and trigger the release of pro- inflammatory cytokines (Tangpukdee et al. 2008).

Although vector control and use of chemoprophylates agents, are common preventive /curative measures against malaria infection;

attention is also being focused on theuse of effective and safe natural remedies as alternative strategies (Fairhurst & Dondorp 2016; WHO 2016). Indonesia has various medicinal plants that

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showed some therapeutic effects. such as (Curcuma xanthorriza Roxb.). It contains msty chemical substances such as flavonoids and In vitro experiments of Curcuma

Roxb. showed some potentials of cueutnin and flavonoid as antioxidant, mttmalanal, and anti-inflammatory. Moreover, they also be used as antibiotic and anti-fungal (Rostdi et al. 2014; Jain, Sood, & GoMhamarajan

As an antioxidant and anti-inflammation, flavoaoids show the ability to bind free radicals and

present the formation of free radicals. As an flavonoid compounds can also the fonnation of proteins, so as to inhibit mm+ial gmwth (Rosidi et al. 2014; Jain, Sood, &

Gouthamarajan 2013; Eva 2015; Rosidi et al.

At the molecular level, curcumin increases intracellular ROS production and inhibits the plaqtodial activity of histone aceyltransferase, the ea;ome needed for chromatin remodeling and transcriptional activation of malaria parasites Sauvain, & Deharo 2011). Although it increase intracellular ROS, curcumin it self also has a low antioxidant effect (Jain, Sood, &

Gouthamamjan 2013). This study therefore is aim«i at investigating the impact of Curcuma

Roxb extract on some haematological (leucocytes and haemoglobin) of PkÄRkIium berghei ANKAinfected mice model.

MATERIALS AND METHODS

ms true laboratory experimental research used aNSt test only randomized control group

&sigm The population of this study were male mice (ALS musculus L.) BALB / c, which were obtaind and treated at the Institute of Tropical Disease Laboratory, Airlangga University.

berghei ANKA (PbA) strain was from the Institute of Tropical Disease University, andwas inoculated in experimettal mice. The ethical research approval obtained from the Research Ethics Commission of Hang Tuah University, Surabaya

& 6/ HC/EC/KEPUHT/2017).

Sample

The samples on this research were male mice esculus L.) BALB / c aged 7-9 weeks.Adult mie (aged 50 days) have better body condition immunological response compared to mice wiå aged 7 weeks (49 days) or > 9 weeks (63

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days). The average weight of mice aged 7-9 wccks ranged from 18 - 25 grams. Male mice have more stable biological condition compared to female mice whose biological condition is affected by the estrous cycle. Adaptation to the environment was carried out for 7 days in all groups of mice and maintained its condition, food and drink. Every six mice were placed in a plastic tub sized 30 x 20 cm that were given chaff mats. All mice were allocated to metabolic cages and kept in an air-conditioned environment (22— 24 oc) with 12-h alternating periods of light/dark and free access to food and fresh water (Nurul 2010; Darmawan 2014).

This study used 5 groups of mice. The sample size was calculated based on the Federer WT formula (k-l) (n-l) 15(Federer 1966). yieldedfive mice for each group, with a total of 25 mice. (1) The first group (G 1) was not infected with PbA and was given Aquadest for four days (Day I — Day 4);

(2) The second group (G2)was infected with PbA and was given Aquadest for four days (Day 1 — Day 4) (3) The third group (G3) was infected with PbA and obtained 150 mg / kg / day extract of curcuma rhizome (Curcuma xanthorriza Roxb.) orally for 4 consecutive days (Day 1 — Day 4); (4) The fourth group (G4) was infected with PbA and obtained

100 mg / kgBW / day extract of curcuma rhizome (Curcuma xanthorriza Roxb.) orally for 4 consecutive days (Day 1 — Day 4); (5) The fifth group (G5) wasinfected with PbA and obtained 50 mg / kgBW / day extract of curcuma rhizome (Curcuma xanthorriza Roxb.)orally for four consecutive days (Day 1 — Day 4).

Inoculation ofP.berghei ANKA

Plasmodium berghei ANKA was one of malaria parasite in rodents, and it had similar biological nature and sensitivity to drugs with malaria species that infected humans, especially P.

falciparum. The blood stage forms ofboth parasites were stored in liquid nitrogen after in vivo passages in BALB/C mice. When the percentage of parasitemia from blood donor mice reached 220%, the bloodwas diluted with pbs, then 200 gl of diluted blood was injected intraperitoneally to all groups of mice. The infected mice were marked and put in a cage according to the treatment group.

Naive mice were infected intraperitoneally (i.p.) with 106 infected red blood cells (iRBC) and their parasitemia level were monitored daily (Blanco et al. 2008; sou4 & Riley 2009;

Widyawaruyanti et al. 2014).

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Curcuma xanthorriza Roxb. Extract

obtained Curcuma xanthorriza Roxb. was

from the traditional market in Surabaya. Extraction process and of the plant taxonomy

was done by the Faculty of Pharmacy, Widya

Mandala University Surabaya (No. 060/ LJ- FF/I/2017). Extraction process was started by washing all fresh rhizome of Curcuma xanthorriza Roxb, then cut into small pieces, and dried. After drying, the pieces were inserted into the milling with a large hole to filter 0.75 mm as preparation

for maceration. A total of 5 kg of fresh rhizome,

macerated with I L of 96% ethanol to get 500 mg extract. Immersion was done for 24 hours while stirring occasionally. The extract produced was filtered with a Buchner funnel to obtain the filtrate,

The residue is re-marred in the same way four

åmes. The extract obtained was then concentrated

with rotary evaporators until the viscous extract was obtained. The extract was left at room

temperature or heated in a low temperature heater to allow the remainder of ethanol to vapor out and

it was extracted into a dry paste (Hutomo &

Winamo, 2005). Preparation of Curcuma xanthorriza Roxb.solution was done by mixing

rhizome extract Curcuma xanthorriza Roxb. with aquadest and CMC-Na solution as a solvent.

The solution was then given to the treatment group every day orally using intragastric sonde.ln this study,the dosagesof Curcuma xanthorriza Roxb. were150 mg / kgBW ofmice/ day, 100 mg / kgBWof mice / day, and 50 mg / kgBWofmice / day. The dose was determined based on the similar study that used 100 mg / kgBW dose of curcuma extract (Widyawaruyanti et al. 2014).

Anesthesia

Animals were anesthetized with ketamine (83mg/g) and xylazine(13mg/g) administered

intraperitoneally for all of the blood

drawingprocedures including tail blood samples.

Parasitemia Examination

Parasitemia was determined daily in all infected groups throughout the study period.

Blood smears were prepared from a drop of blood

of the animal's tail on slides and stained with

Giemsa. The slides were allowed to dry and then fixed with methanol. After fixing, the slides were allowed to dry and then stained with 10% Giemsa in methanol for 30 minutes. The slides were then

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rinsed with water and then allowed to dry percentage of parasitemia was describCd

number of parasitized red blood cells (PRE(

1,000 erythrocytes on at least ten

microscopic fields. The

percentage parasitemia was calculated using the followJr

pumber of infected erythrocytes X 100 1000 erythrocytes

Blood Examination

Blood tests to determine the number

leukocytes and hemoglobin performed on dayof after the mice terminate. Blood sampling was _done

on the heart of mice, following these

_protocol:

animal positioned on its back, the _abdomen

wasopened and the heart visualized through the

membranous part of thediaphragm. A 21 _gauge needle and 3cc syringe were used to obtain 0.5 ml of blood from the left ventricle. The blood was then immediately transferred to 20ml capillary _pipettes containing the appropriate anticoagulants for cell counting. EDTA anti-coagulated blood _samples were used to obtaina complete blood count with a Mindray Auto Hematology Analyzer.

Statistical Analysis

The one way ANOVA test was used to analyze

and compare the results at a 95% confidence level if normality and homogenity were obtained. If the data were not met the parametric criteria, the data

were then analyzedusing Kruskal Wallis

^test, followed by the Mann-Whitney U test at = 0,05.

RESULTS

Acute toxicity test

The experimental mice ingested

^with extract of rhizome of Curcuma xanthorriza ^Roxb.

in all doses did not show any indication of

^gross physical or behavioral changes, such as hair erection, reduction in feeding and motor activities,

weight loss, lacrimation, diarrhea, depression or abnormal secretions within 24 hour of ^monitoring period. No fatalities occurred within the four days observation period.

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Level Parasitemia

Normality test results indicate that the normality requirement is not fulfilled because p <

0,05. This study was using non parametric analysis

Kruskal Wallis to analyze the difference of % parasitemia between the groups.

PARASITEMIA LEVEL %

7

_6.14

3.03 2.83 4

1.97 2.0

7

1.15 .56

7

* 0. _PdB•15

0

DO DI

DAYS

Figurel. Changes Level Parasitemiaof Plasmodium berghei ANKAInfected & Non Infected Mice With Rhizome Extract Of Curcuma xanthorriza Roxb

GI : not infected with PbA and was given Aquadest for four days

G2 : infected with PbA and was given Aquadest for four days (Day 1 — Day 4)

G3 :infected with PbA and obtained 150 mg / kg / day extract of curcuma rhizome (Curcuma xanthorriza) Roxb.)orally for 4 consecutive days (Day 1 — Day 4)

G4 : infected with PbA and obtained 100 mg / kgBW / day extract of curcuma rhizome (Curcuma xanthorriza Roxb.) orally for 4 consecutive days (Day 1 — Day 4)

G5 : infected with PbA and obtained 50 mg / kgBW / day extract of curcuma rhizome (Curcuma xanthorriza Roxb.)orally for four consecutive days (Day 1 — Day 4).

Level of parasitemia in DO showed that there was no significant difference (p > 0,05), Gl did not show parasite growth because the mice in this group were not infected with PbA and got aquadest. Mean value of % parasitemia on DI — D4 between groups ofthis research showed that G2 (infected with PbA and was given Aquadest for four days) has the highest % parasitemia and G3 (infected with PbA with 150 mg/KgBW extract curcuma) has th e lowest % parasitemia.

Leucocyte Count

Normality test is not fulfilled because p >

0,05, we used a nonparametric analysis Kruskal Wallis to analyze the difference of leucocyte count between the groups. Haematological changes

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associated withadministration ofrhizome extract of Curcuma xanthorriza Roxb to Plasmodium berghei ANKAinfected mice were investigated.

The effects rhizome of Curcuma xanthorriza Roxbextractto infected and uninfected mice on leucocyte count are shown in Table l.

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Table 2. In

Level Table l. Changes In LajcocyteCount of

Plasmodium berghei ANKA Infected & Non Infected Mice With Rhizome Extract Of Curcuma

xanthorriza Roxb

Changes Hemoglobin

Plasmodium berghei ANKAInfected & NOf Infected Mice With Rhizome Extract Of

Group Mean (/gL)

4620

16980

21280 28600 30700

Std.

Deviation 931

1881

2705 2026 2328

Grou Mean /dL

11,51 1,36 11,28

8,80 10,08

o,lS9

0,204 1,268 0,803 o, 120

Descriptive and statistical analysis above shows that there are significant differences in the groups. The lowest leukocyte count was found in Gl (4620) and the highest leukocyte count was in

G5 (30700). Based on Mann whitney post hoc

analysis showed that the number of leukocytes between groups ( Gl with G2 — G5; G2 with G4 — G5; G3 with G4 — G5) was significantly different (p <0.05), but there were no significant differences between groups G2 with G3 and G4 with G5 (p>

0,05).Several treatnent groups were found to have sigiificant differences in leukocyte counts among other treatment groups. This may be due to differences in the administration of extracts with different doses and containing different levels of bioactive components as well.

Hemoglobin Level

The data of hemoglobin level shows normal distribution, so the analysis used is non parametric analysis, Kruskal Wallis. Haematological changes associated with administration of rhizome extract

of Curcuma xanthorriza Roxb to Plasmodium

berghei ANKA infected mice were investigated.

The effects rhizome ofCurcumaxanthorriza Roxb extract to infected and uninfected mice on hemoglobin level are shown in Table 2 :

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003 <a a a 0,05)

Descriptive analysis and statistics in the table above shows the difference in the hemoglobin level. The lowest hemoglobin level was found in G4goup (8>80) and the highest hemoglobin levelwas in Gl group (11,51). Based on Mann whitney post hoc analysis showed that the number of hemoglobin level between groups ( Gl with G2

— G5; G2 with G4 —G5) was significantly different (p <0.05), but there were no significant differences between groups G2 with G3; G3 with G4- G5; and G4 with G5 (p> 0,05). Several treatment groups were found to have significant differences in hemoglobin level among other treatment groups.

This may be due to differences in extract doses and containing different levels ofbioactive components as well.the other cause of the difference is also due to the development of malarial parasites in erythrocytes.

DISCUSSION

High level parasitemia infections are often associated with malaria severity which triggers a defense mechanism that inhibits parasite multiplication and an important factor in the survival of the host. The chemical content of curcuma (Curcuma xanthorrhiza Roxb) includes

flavonoids and curcumin has an anti-tumor,

antioxidant, and antiplasmodium roles. Curcumin has two roles both anti oxidant and pro oxidant activities. Curcuma has anti-malarial activity through inhibition of the parasitic hystone acetyltransferase enzyme which plays a role in the transcription process and also through increased production of ROS in parasites. ROS can increase the expression of CD 36 on the surface of monocytes / macrophages, which facilitates the process of nonopsonic phagocytosis of infected erythrocytes. In addition to the antimalarial effects

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of curcuma, this plant also has the effect of regulating the immune response (immunomodulators) in various immune cells (Mimche, Taramelli, & Vrvas 2011; Jain, Sood, &

Gowthamarajan 2013)

Based on the results of statistical analysis on the number of leukocytes it can be seen that G3 (mice group given a dose ofcurcuma extract 150 mg/ KgBW) showed a lower leukocyte count than G4 (mice group given 100 mg / KgBW) and G5 (group of mice given a dose of curcuma extract of 50 mg/ KgBB). Leukocytes count in the G3 (mice group given a dose of curcuma extract 150 mg / KgBW) did not show a siglificant difference in the G2 (the group of mice inoculated with P. berghei and received aquadest). The negative control group had significantly lowest leukocyte counts compared to other groups infected with Plasmodium. This result caused by existence of erythrocytes containing Plasmodium triggers the immune system's initial response to eliminate Plasmodium, which is played by leucocyte cell such as macrophages, neutrophils and lymphocyte I)scriptive analysis showed that the positive control group (G2) had a lower leukocyte count than all treatnent groups that received curcuma extract (G3, G4 and G5), although statistically the number of leukocytes between G2 and G3 was not significantly different. Some treatment groups on this study were found to have significant differences in leukocyte counts among other treatment groups. This might occur because of differences in the administration of extracts with different doses and containing different levels of bioactive components.

The groups that received curcuma extracts were found to experience a drastic increase in leukocyte counts probably due to ongoing parasite growth which triggered the work of phagocytic cells and curcumin activity was effective against increased leukocyte counts (Reddy et al. 2005;

Jain, Sood, & Gowthamarajan 2013). Bioactive components contained in herbal medicines such as curcuma can activate the RNA and protein synthesis pathways that stimulate proliferation of

cells (neutrophils, eosinophils, monocytes, and lymphocytes) (Reddy et al. 2005;

Jain, Sood, & GoMhamarajan 2013). The increase in the number of leukocytes in the group that received the extract was accompanied by a decrease in the level of parasitemia. The lowest reduction in parasitemia was found in the group

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who received a dose of 150 mg / KgBW (G3) compared to the group who received other doses.

In this study it can be concluded that administration

of curcuma extract will increase the number of

leukocytes associated with activity to phagocyte infected erythrocytes. In the positive control goup there was an increase in leukocytes caused by the malaria infection process, but the increase was lower than the group that received the extract. This is probably due to the absence of Curcuma extract, so that leukocyte cell proliferation did not occur as much as the group that received Curcuma extract.

Based on the results of hemoglobin examination showed that the group infected with PbA showed a decrease in hemoglobin compared to the negative control group that was not infected.

The decrease in hemoglobin count can be caused by two main factors such as increased erythrocyte destuction and decreased erythrocyte production.

Increased destruction of erythrocytes is caused by:

(1) rupture of pRBC (parasitized RBC); (2) phagocytosis of erythrocytes, either erythrocytes containing parasites or erythrocytes that have not been infected; (3) hypersplenism, contributes to fre initial phase of acute malaria by eliminating parasites from erythrocytes without lysis or ingestion of erythrocytes with a shorter life span of

the erythrocytes; and (4) autoimmme

extravascular hemolysis (Perkins et al. 2011) The decrease in hemoglobin level in the group that received the extract was greater than the positive control, although the difference was not significant between positive control (G2) and G3 and G5. This is because the active ingedients in Curcuma extract have antimalarial activity that can increase immune cell activity, especially phagocytic cells and the production ofROS, which can reduce the number of erythrocytes and hemoglobin (Reddy et al. 2005; Haddad, Sauvain,

& Deharo 2011). This allegation is supported by data on decreasing levels of parasitemia in the group that received curcuma ext-act. The group that received a dose of 150 mg / KgBW (G3) showed the lowest level of parasitemia compared to other groups and a hemoglobin level that was almost the same as positive control.

CONCLUSION

Based on the results of the statistical data analysis test and the interpretation of the results of the study, it can be concluded that:

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l, Plasmoditun bqhei ANKA infection can reduce hemoglobin md incrcasc leuaxyte count in BALB / c malc mice (Mus musculus L).

2. administration of ginger rhizome extract (Curcuma xanthotyhiza Roxb) Has an effect on increasing leukocytes and decreasing hemoglobin in male BALB / c mice (Mus musculus L.) inoculated with Plasmodium berghei ANKA 3. administration of temuiawak extract (Curcuma xanthorrhiza Roxb.) At a dose of 150 mg

/ KgBB is the most effective dose due to the

increased inhibitory effect of parasitemia of BALB / c male mice (Mus musculus L.) infected with Plasmodium berghei ANKA.

Acknowledgement

This research can be held because of funding support from LPPM and the Faculty of Medicine, University of Hang Tuah

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The ICOME 2018

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ISBN: 978-623-93960-1-5

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PDF iC*ME - dspace.hangtuah.ac.id

iC*ME

CURRENT RESEARCH

ON HYPERBARIC, UNDERWATER AND COASTAL MEDICINE

needing noo/ø

Current Research on Hyperbaric, Underwater and Coastal Medicine

Theme:

Hyperbaric and Underwater Medicine for the Future

needing Zoo/e

27-28 July 2018 Four Points Hotel Surabaya, Indonesia

FAKULTAS KEDOKTERAN UNIVERSITAS HANG TUAH

Table of Contents

lc ME

The Effect of Curcuma (Curcuma xanthorrhiza Roxb.) Rhizome Extract to

Amount of Leukocytes and Haemoglobin in Male BALB/c Mice

(Mus musculus L.) Infected by Plasmodium berghei ANKA

& 6/ HC/EC/KEPUHT/2017).

dc ME

was done by the Faculty of Pharmacy, Widya

for maceration. A total of 5 kg of fresh rhizome,

The residue is re-marred in the same way four

with rotary evaporators until the viscous extract was obtained. The extract was left at room

it was extracted into a dry paste (Hutomo &

Winamo, 2005). Preparation of Curcuma xanthorriza Roxb.solution was done by mixing

Anesthesia

intraperitoneally for all of the blood

Parasitemia Examination

of the animal's tail on slides and stained with

rinsed with water and then allowed to dry percentage of parasitemia was describCd

1,000 erythrocytes on at least ten

Blood Examination

Blood tests to determine the number

on the heart of mice, following these

wasopened and the heart visualized through the

were then analyzedusing Kruskal Wallis

RESULTS

The experimental mice ingested

in all doses did not show any indication of

iC&ME

PARASITEMIA LEVEL %

7

0

DAYS

G5 (30700). Based on Mann whitney post hoc

of Curcuma xanthorriza Roxb to Plasmodium

003 <a a a 0,05)

flavonoids and curcumin has an anti-tumor,

lc*ME

of curcuma extract will increase the number of

the erythrocytes; and (4) autoimmme

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