SERBA-SERBI PERJURNALAN

Setelah bercakap-cakap dengan ibu Ati (staff Althea di lt. 3 eijkman), ini ada beberapa aturan jurnal yang mungkin terlewat oleh teman-teman .Thanks to Sabrina buat jurnalnya ( Dia udah bener) jadi bisa jadi pedoman temen-temen dalam membuat jurnal.

Beberapa aturan yang mungki terlewat adalah 1. Halaman 1 berisi judul dan data lengkap author (penulis, dombig 1 dan 2) . Bebas tulisan dan spacingnya yang penting 1 halaman. Bukan school of medicine tapi faculty of medicine, bukan padjadjaran university tapi universitas Padjadjaran. 2. Halaman 2 abstract. Judul ditulis lagi diatas abstract. Tulisan abstractnya rata kiri yaaa. Keyword ditulis dipisah key word dan disusun secara alphabet. Untuk abstract spacing tidak ditentukan jadi bebas. Abstract berisi : background, methods, results dan conclution 3. Semua sub judul : introduction, methods, results dan disccussion ditulis rata kiri. 4. Table dan figure maximal 6 ! 5. Untuk citasi harap diperhatikan. Keterangan ada dibawah. 6. References maximal 20 dan textbook tidak boleh lebih dari 20% references. Misalnya references ada 15 jadi textbook yang digunakan tidak boleh lebih dari 3, klo references ada 20, textbooknya 4. Jika ada textbook yang tahun terbitnya sudah lebih dari 10 tahun sebaiknya hubungi pihak althea boleh dimasukan apa tidak. 7. Maximal 20 halaman mulai dari judul sampai akhir references 8. Dan ada Attachment , di file yang berbeda dengan jurnal berisi semua table dan figure dan keterangnnya yang ada di jurnal. Tidak udah memakai page numering. 9. Gunakan word 97-2003 atau 2010.

Temen-temen tolong baca lagi aturan di website althea nya ya, jangan cuma dari printout slide waktu kita sosialisasi. Karna ada beberapa aturan yang tidak tertulis di printout itu.

Proses ketika kita memasukan jurnal ada 2 kali pengcheckan sampai surat submission keluar. Pertama : check format ( apakah sudah sesuai dengan aturan apa belum), jika belum maka kita akan diemail lagi untuk direvisi. Setelah format benar, ada check content ( yang dilihat keseimbangan antara introduction, method, results dan dissccusion. Misalkan introduction panjang maka tim redaksi althea akan meminta untuk mengurangi isinya, maka kita harus ke dombing lagi untuk bersama-sama mengurangi introductionnya jadi dombing harus tau jika ada perubahan)

Nah, proses dari mulai kita kirim email sampai dikeluarkannya surat submission sekitar 7-10 hari. (tergantung banyaknya kesalahan kita atau banyaknya jurnal yg masuk). Jadi ibu Ati berpesan kalau bisa diurus dari sekarang biar ga numpuk diakhir-akhir pengumpulan, kalau numpuk waktu yang diperlukan semakin lama

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Jika semua sudah okee, kita akan mendapatkan email jika surat sudah bisa diambil ( surat submission yang telah ditanda tangani dr. Sri Endah).

Waktu sosialisasi kita diberi tahu untuk print sendiri lalu datang ke dr. Sri endah, ini berubah. Jadi kita akan mendpatkan email jika surat sudah bisa diambil . perubahan ini karena tim althea belum siap dengan proses yang mengirimkan email surat submission ke kita (aplikasinya belum berjalan dengan baik)

Oia, ada 2 cara untuk revisi : Pertama kita kirim ke email chiefeditor_amj@aimejournal.org Atau kita datang langsung ke eijkman lt. 3 ruangan MKB ( majalah kedokteran bandung) di sana ada Ibu Ati yang siap sedia memeriksan jurnal kita secara langsung.

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Semangaat teman-teman !

Bixa orellana leaf infusion as an antiinflammatory agent in carrageenaninduced wistar rats

Sabrina Munggarani Yusuf, 2Enny Rohmawaty, 3Rama Nusjirwan 1Faculty of Medicine, 2Department of Pharmacology and Therapy, 3Department of Surgery, Faculty of Medicine, Universitas Padjadjaran/General Hospital Hasan Sadikin, Bandung
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1. Sabrina Munggarani Yusuf, Faculty of Medicine, Universitas Padjadjaran, Jalan Raya Bandung Sumedang Km21, Jatinangor, Sumedang. Phone: +6281513130450 Email: sabrinamyusuf@yahoo.com

1. Enny Rohmawaty, dr., M.Kes, Department of Pharmacology and Therapy, Faculty of Medicine, Universitas Padjadjran/General Hospital Hasan Sadikin, Jalan Pasteur No. 38 Bandung 40161 Phone : +628157133135 Email :

2. Rama Nusjirwan, dr., SpBTKV, Department of Surgery, Faculty of Medicine, Universitas Padjadjaran/General Hospital Hasan Sadikin, Jalan Pasteur No.38, Bandung 40161. Phone : +628159972766

Bixa orellana leaf infusion as an antiinflammatory agent in carrageenaninduced wistar rats

Abstract

Background: One of the characteristics of inflammation is swelling or edema. Inflammation can be managed by traditional medicine, such as Bixa orellana. Bixa orellana leaf is found to contain flavonoid and tannin responsible for its antiinflammatory effect. This study was to analyse the ability of Bixa orellana leaf infusion (BOLI) to supress paw edema in carrageenan-induced rats. Methods: Thirty female Wistar rats used were randomly divided into five groups. Negative group was given 5 mL aquades, three groups received BOLI with 0,09 g; 0,18 g; and 0,36 g dosage respectively; and positive group was given 0,9 mg diclofenac. At 1 hour after treatment, all rats were induced by carrageenan injection subcutaneously. Paw edema changes were quantified at 0, 1, 2, 3, 4, 5, 6, and 24 hour afterwards. Results: Based on paw edema inhibition percentage, 0,18 g of BOLI was shown most effective (16,9744%) compared to 0,09 g (10,9610%) and 0,36 g (7,5087%). Data were analysed using Kruskal-Wallis and Mann-Whitney test. No significant differences of antiinflammatory effect were observed between groups that were treated with 0,18 g of BOLI and diclofenac (p > 0,005). Conclusions: BOLI can supress inflammation comparable to diclofenac. The effective dosage is 0,18 g/200 g BW/day. Key words: antiinflammatory, Bixa orellana leaf, paw edema

Introduction Inflammation is a protective response invoked in organisms towards alteration in normal tissues. It has five main characteristics, which are rubor (redness), tumor (swelling), calor (heat), dolor (pain), dan functio laesa (loss of function). When uncontrolled, diseases can follow. To name a few, chronic inflammation can cause rheumatoid arthritis, atherosclerosis, and tuberculosis. Thus, antiinflammatory drugs, both steroidal and non-steroidal, play a pivotal role in taking the response under control.1 However, unwanted side effects such as obesity and tummy cramps encourage the escalating demands for herbal medicine which is generally considered safer than many other conventional drugs. 2 Bixa orellana, lipstick tree from Bixaceae family, has been approved safe for human consumption by Food and Drug Administration (FDA) of The USA It is beneficial in treating fever, stomachache, sorethroat, burn injury, and epilepsy pertaining to its antiinflammatory effect.3 Previous study by Y. Yoke Keong et al (2011) using Bixa orellana leaf extract proved flavonoid and tannin in favor of that advantage. Moreover, there are some other chemical compounds found within Bixa orellana leaf, for instance sterol, saponin, and ishwarane.4 This study was undertaken to analyse the antiinflammatory effect of Bixa orellana leaf infusion (BOLI) for an easier home-based production. Paw edema in carrageenaninduced rats were investigated. Efficacy compared to diclofenac and most effective dosage were then determined.

Methods This experiment was carried out in Animal Laboratory of Department of Pharmacology and Therapy in General Hospital Hasan Sadikin Bandung in October 2012. Bixa orellana leaves were procured from Lembang, Bandung, and were botanically identified at the Herbarium Jatinangor, Universitas Padjadjaran. Thirty healthy female Wistar rats (2-3 months old) weighing 200 50 g were used as subjects. All rats were housed in homogenous temperature and dark-light cycle for 7 days with ad libitum access to food and drink. The rats were subsequently fasted for the next 24 hours to promote optimal inflammatory response. The procedure was done in accordance with Russel and Burchs principle of humane experimental techniques: Reduction, Refinement, Replacement (3R).5 BOLI was prepared from dried Bixa orellana leaves weighed according to dosage and 100 mL of water for every treatment group. Mixed raw material was heated in an infusion pan for 15 minutes after the temperature reached 90C while being stirred every 5 minutes. Next, BOLI was filtered using flanel fabric. Thirty milliliters of BOLI were used for each rat in the treatment group receiving as much as 5 mL. Extra water were added to gain BOLI volume as required.6 At one hour before treatment, basal right paw volume was quantified with a pletysmometer until lateral maleolus which was marked with a label sticker. Models were randomly divided into five groups. Negative group was given 5 mL aquades, three groups received BOLI with 0,09 g; 0,18 g; and 0,36 g dosage respectively; and positive group was given 0,9 mg diclofenac. All treatments were given orally. An hour afterwards, edema was induced by

carrageenan injection subcutaneously in subplantar area of the posterior right paw. In order to alleviate the pain, all rats were locally anesthesized using 20 mg/kg BW of ketamine hydrochloride intraperitoneally prior to subcutaneous injection. Paw edema changes were determined at 1, 2, 3, 4, 5, 6, and 24 hour after treatment. Moreover, the efficacy of each treatment group to supress paw edema when compared with negative group was calculated. Results were classified into stadium 1 (% edema inhibition < 10% or poor), stadium 2 (% edema inhibition 10-15% or moderate), and stadium 3 (% edema inhibition > 15% or excellent). Data were statistically analysed using Kruskal-Wallis and Mann-Whitney nonparametric tests because of the anomalies in the distribution of the data. P value 0,05 showed significant difference.

Results Paw volume for each time interval compared to basal paw volume can be seen in Table 1. Negative group had the largest paw edema changes at 3 hour after treatment (1,3667 mL). Meanwhile, the smallest paw edema changes were seen in group of rats given 0,18 g of BOLI at 24 hour after treatment (0,8333 mL).

Table 1. Paw Edema Changes at Every Time Points

Group V0 V1 1 0,66 1,15 2 0,61 1,05 3 0,56 1,00 4 0,61 1,01 5 0,68 1,00 *SD: Standard Deviation V2 1,28 1,16 1,01 1,10 1,00 V3 1,36 1,25 1,16 1,30 1,20 V4 1,31 1,13 1,06 1,26 1,18 V5 1,33 1,11 1,11 1,23 1,11 V6 1,25 1,08 0,98 1,18 1,05 V24 0,96 0,90 0,83 0,91 0,90 Mean + SD* 1,16 1,03 0,96 1,07 1,01

Mean Paw Edema Changes

1.4000 Paw Edema Volume (mL) 1.2000 1.0000 0.8000 0.6000 0.4000 0.2000 0.0000 1 2 3 Group 4 5 1.1667 1.0396 0.9688 1.0792 1.0167

Figure 1. Mean Paw Edema Changes

From the figure above, group given 0,18 g of BOLI had the smallest mean paw edema changes. Data were then conversed into percentage. Results were shown in Figure 2. Paw volume in all groups gradually increased at 1 to 3 hour time point, then started to fall afterwards. This proved the efficacy of carrageenan to optimally induce edema-forming effect at the first two hours after injection, after which was inhibited by both treatment groups and positive group.

Paw Changes Percentage

120.0000 100.0000 80.0000 60.0000 40.0000 20.0000 0.0000 V1 V2 V3 V4 V5 V6 V24 Time Points (hour) Paw Changes (%) Negative group 0,09 g of BOLI 0,18 g of BOLI 0,36 g of BOLI Positive group

Figure 2. Paw Changes Percentage at Every Time Points

Kruskal-Wallis test was exerted to identify the effect of various dosage of BOLI towards mean paw volume. Paw edema was significantly supressed at 2 and 6 hour after treatment (p < 0,05).

Table 2. Kruskal-Wallis test

Test Statistics V1 Chi-Square Df Asymp. Sig. 5.938 4 .204 V2 13.395 4 .009 V3 7.911 4 .095
a,b

V4 8.964 4 .062

V5 7.144 4 .128

V6 10.212 4 .037

V24 2.439 4 .656

a. Kruskal Wallis Test b. Grouping Variable: Group

To investigate antiinflammatory effect between groups, the author used Mann-Whitney test. When compared to negative group, significant differences were not shown solely in group given 0,09 g of BOLI. In addition, significant difference among three dosages of BOLI (p < 0,05) was merely seen between groups given 0,18 g and 0,36 g of BOLI at 4 and 6 hour time points. Paw edema changes in treatment groups and positive group were then compared to negative group. Results yielded paw edema inhibition percentage depicted in Figure 3. Group treated with 0,18 g of BOLI had the biggest inhibition (16,9744%) followed closely by diclofenac (13,7985%). Both groups didnt show significant differences when compared using Mann-Whitney test.

Mean Edema Inhibition Percentage

18.0000 16.0000 14.0000 12.0000 10.0000 8.0000 6.0000 4.0000 2.0000 0.0000 16.9744 13.7985 10.9610 7.5087 Edema Inhibition (%)

3 Group

Figure 3. Mean Edema Inhibition Percentage

Discussion Antiinflammatory effect in rats was examined by calculating supression of paw edema as the simplest chemical method in Animal Laboratory of Department of Pharmacology and Therapy. Carrageenan was handpicked as the inducer of inflammation due to its acute, non-immune, well-studied, and easily produced response. The efficacy of carrageenan shown in Figure 2 is related to its biphasic course of reaction. The first phase is mediated by histamine, serotonin, 5hydroxytryptamine, and bradykinin release lasting for 2 hours.7 The inability of Non-steroidal Antiinflammatory Drugs (NSAIDs), such as diclofenac, to intervene is because this phase doesnt comprise the activation of cyclooxygenase (COX) pathway. On the contrary, the second phase being manipulated is carried out by prostaglandin, protease, and lysosome production taking place as long as 35 hours, or even 24 hours after injection.8 Enhancement of paw edema at 1-3 hour time interval was also observed in the positive group. Due to its half life (1-2 hours), diclofenac attains optimal

antiinflammatory effect at 2-3 hours after oral administration. It supresses nitric oxide (NO) by altering the uptake and release of fatty acid responsible for the syntesis of arachidonic acid, the precursor of COX.2,9 Identical response also occured in the treatment groups. This can be explained by the ability of chemical constituents in BOLI, flavonoid and tannin, to block the second phase of carrageenan-induced inflammatory reaction. While flavonoid inhibits the production of COX-2 and inducible nitric oxide synthase (iNOS), tannin helps decrease reactive oxygen species (ROS) and free radicals concentration in the blood through its oxidation.10-12 Tannin is also capable of forming complex with protein, polysaccharide, alkaloid, nucleic acid, and mineral.13 Therefore, further study should be conducted to examine the effect of tannin administration topically to accelerate wound healing. Paw edema at every time intervals were analysed using Kruskal-Wallis test. Different dosages of BOLI showed significantly different antiinflammatory effect at 2 and 6 hour. After administration, flavonoid metabolites obtain maximal serum concentration at 1 hour and last for 4 hours. During that period, flavonoid concentration in blood plasma is minimally altered.14 Factors that affect the time required for flavonoid to achieve its peak serum concentration include variation of food matrix and metabolism enzymes, causing disparities among efficacy in polyphenol absorption.15 On the other hand, it was assumed that the elimination half life of BOLI was 6 hours after treatment with carrageenan. Mann-Whitney test results showed no significant differences between group with 0,09 g of BOLI and negative group. This happened because this dosage was insufficient to attain optimal serum concentration.14 When comparing

effects among treatment groups, only group with 0,18 g and 0,36 g of BOLI displayed significant difference at 4 and 6 hour after treatment. According to Figure 1 and 3, group with 0,18 g of BOLI was most effective. Inefficiency of 0,36 g of BOLI might have occured due to the waning of its antiinflammatory effect. However, this assumption should be verified by further study. Mean paw edema inhibition percentage clearly shows efficacy when groups are compared. No significant differences were found between 0,18 g of BOLI and positive group. Thus, 0,18 g of BOLI is able to prevent paw volume from increasing which is comparable to diclofenac, rating stadium 3. On the contrary, group with 0,09 g of BOLI was classified as stadium 2 and group with 0,36 g of BOLI was inclusive to stadium 1 as an antiinflammatory agent. These results provided scientific evidence for the benefit of Bixa orellana leaf use to counter disruptive effect of inflammation, in harmony with the prior study conducted by Y. Yoke Keong et al (2011) using extract of Bixa orellana leaf. Bixa orellana leaf in the form of both extract and infusion showed comparable antiinflammatory effect because the ratio between the plant and the diluent doesnt have significant impact to the chemical compounds contained within. In fact, chemical compositions in the leaf are mainly dependent on the temperature and duration of the extraction process.16 As a conlusion, present data suggested that Bixa orellana leaf infusion has an antiinflammatory effect comparable to diclofenac. The most effective dosage is 0,18 g/200 g BW/day. Studies in the near future should clarity its potential as a topical antiinflammatory agent as well as its probable toxic effect.

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