ANTIARTHRITIS ACTIVITY OF COMBINATION OF RED GINGER (Zingiber officinale Rosc.) AND ACALYPHA (Acalypha indica Linn) EXTRACTS ON ARTHTRITIS RAT Aktivitas Antiarthritis Kombinasi Ekstak Jahe Merah (Zingiber officinale Rosc.) dan Acalypha (Acalypha indica Li

  

ANTIARTHRITIS ACTIVITY OF COMBINATION OF RED GINGER

(

Zingiber officinale Rosc.) AND ACALYPHA (Acalypha indica Linn)

EXTRACTS ON ARTHTRITIS RAT

  

Aktivitas Antiarthritis Kombinasi Ekstak Jahe Merah (Zingiber officinale Rosc.) dan

Acalypha (Acalypha indica Linn) Terhadap Tikus arthritis

Rina Herowati, Gunawan P. Widodo and Ismi Puspitasari

  

Faculty of Pharmacy, Universitas Setia Budi

Jalan Letjen Sutoyo Surakarta 57127, Indonesia

e-mail:rinagunawan53@gmail.com

  

ABSTRAK

Arthritis adalah penyakit peradangan kronis yang mempengaruhi sistem muskuloskeletal dan disebabkan oleh

reaksi autoimun multisistem. Beberapa tanaman obat bekerja secara sinergis untuk mengurangi peradangan sendi

kronis arthritis. Tujuan dari penelitian ini adalah untuk mengetahui aktivitas antiarthtritis kombinasi rimpang jahe

merah dan Acalypha seluruh tanaman ekstrak etanol pada lengkap Freund adjuvant diinduksi arthtritis. Arthritis

ajuvan diinduksi pada tikus pada hari 0. Sembilan hari setelah inokulasi, hewan-hewan itu dipilih dan didistribusi-

kan ke dalam 5 kelompok. Satu kelompok diberi 0,5% larutan CMC sebagai kelompok kendaraan yang diobati, yang

lain diberi ekstrak jahe merah (280 mg/kg bb), ekstrak Acalypha (200 mg/kg) kombinasi merah jahe dan ekstrak

Acalypha (140 mg/kg bb : 100 mg/kg bb), dan triamcinolone (0,3 mg/kg bb) secara lisan. Tingkat peradangan di-

evaluasi oleh belakang sweeling kaki, didukung oleh histopatologi sendi pergelangan kaki. Hasil penelitian menun-

jukkan bahwa sediaan oral ekstrak Acalypha, ekstrak jahe merah, serta kombinasi mengurangi mereka belakang

kaki bengkak, sebanding dengan yang dari triamsinolon. Studi Histopatologi mengungkapkan penurunan yang sig-

nifikan dalam infiltrasi mononuklear dan meningkatkan ruang sendi pergelangan kaki. Kombinasi rimpang jahe

merah dan ekstrak acalypha menunjukkan efek perlindungan terhadap pembantu diinduksi arthritis.

  Kata kunci: rimpang jahe merah, Acalypha, Freund adjuvant lengkap, antiarthritis, histopatologi.

  

ABSTRACT

Arthritis is a chronic inflammatory disease affecting the musculoskeletal system and caused by a multisystem au-

toimmune reaction. Some medicinal herbs work synergistically to reduce chronic joint inflammation in arthritis.

  

The purpose of this study was to determine the antiarthtritis activity of combination of red ginger rhizome and

acalypha whole plant ethanolic extracts on complete Freund’s adjuvant induced arthtritis. Adjuvant arthritis was

induced in rats on day 0. Nine days after inoculation, the animals were selected and distributed into 5 groups. One

group was given 0.5% CMC solution as vehicle-treated group, the others were given red ginger extract (280 mg/

kg bw), acalypha extract (200 mg/kg bw) combination of red ginger and acalypha extracts (140 mg/kg bw:100

mg/kg bw), and triamcinolone (0.3 mg/kg bw) orally. The degree of inflammation was evaluated by hind paw

sweeling, supported by histopathology of ankle joints. Results suggested that oral dosage of acalypha extract, red

ginger extract, as well as their combination reduced hind paw swelling, comparable to that’s of triamcinolone.

  

Histopathology study revealed the significant reduction in mononuclear infiltration and increased the ankle joint

space. The combination of red ginger rhizome and acalypha extracts showed protective effect against adjuvant

induced arthritis.

  Keywords: red ginger rhizome, acalypha, complete Freund’s adjuvant, antiarthritis, histopathology

  INTRODUCTION

  Rheumatoid arthritis (RA) is a chronic, progressive, systemic inflammatory disorder affecting the synovial joints and typically pro- ducing symmetrical arthritis that leads to joint destruction, which is responsible for the defor- mity and disability. The consequent morbidity and mortality has a substantial socioeconomic impact. The world prevalence of rheumatoid arthritis is about 0.5-1 % of the population. It usually occurs in the people between 25 and 55 years of age. Women are affected more of- ten than men at ratio of 3 to 1 (Patwardhan et

  al ., 2010).

  Adjuvant induced arthritis (AIA) in rats is a chronic inflammatory disease character- ized by infiltration of synovial membrane in association with destruction of joints resem- bles RA in humans. RA progress was divided in three stages. The first stage is the swelling of the synovial lining, causing pain, warmth, stiffness, redness and swelling around the joints. Second is the rapid division and growth of cell, or pannus, which causes the synovium to thicken. In the third stage, the inflamed cell releases enzyme that may digest the bone and cartilage, often causing the joints to loses its shape and alignments, more pain and loss of movements. The most commonly prescribed medication for RA treatment is steroidal, non- steroidal anti-inflammatory, disease modifying antirheumatic and immunosuppressant drugs.

  Though the goal of these drugs have been to relieve pain and to decrease joint inflamma- tion, to prevent joint destruction and to re- store function of disabled joints, these drugs are known to produce various side effects in- cluding gastrointestinal disorders, immunode- ficiency and humoral disturbances. According- ly, reducing side effects should be considered while designing improved therapeutics for Rheumatoid arthritis, besides enhancing me- dicinal effectiveness. Traditional treatment is being increasingly recognized as an alternate approach to arthritic treatment (Bansod et al.,

  Red ginger (Zingiber officinale Rosc.) has been used for medicinal purposes since antiquity. In particular, it has been an im- portant plant for the traditional Indonesian medicines. Although one of its indications has been historically to treat rheumatic disorders and although ginger extracts have shown the ability to inhibit arachidonic acid metabolism and have antiinflammatory action and/or an- tirheumatic properties, there are very limited published reports on the efficacy of this plant (Funk et al

  ., 2009; Rehman et al., 2011; Shen et al., 2003). Acalypha indica belongs to Euphor- biaceae family and can be found in plains of Indonesia. It has been traditionally used as an- thelmintic, cathartic, scabies, and rheumatism. The extracts of A. indica have been reported to possess anti-inflammatory, analgesic and anti- microbial properties (Krishna et al., 2011).

  The objective of our study was to de- termine the antiarthtritis activity of combina- tion of red ginger rhizome and acalypha whole plant ethanolic extracts on Complete Freund’s adjuvant (CFA) induced arthtritis.

  MATERIAL AND METHODS Materials

  The fresh plants of A. indica were col- lected from the local region and rhizomes of red ginger were purchased from local vendor. The plant materials were identified and au- thenticated taxonomically. CFA (Sigma) con- taining suspension of 0.1% w/v killed Myco- bacterium tuberculosis bacteria homogenized in liquid paraffin. Wistar rats were used for the study. The ani- mals were acclimatized to animal house condi- tions. The rats were fed with commercial rat’s diet and water ad libitum.

  Methods Preparation of extracts

  The rhizomes of red ginger were washed, cut into small pieces, and dried under shade. Coarse powder was made and extracted

  Rina Herowati, Gunawan P. Widodo and Ismi Puspitasari by maceration with 90% ethanol for 5 days at room temperature. Dried plants of A. indica were crushed into powder and extracted by maceration with 90% ethanol for 5 days at room temperature. The whole extract of indi- vidual plants was collected in conical flasks, filtered and the solvents were evaporated to dryness under reduced pressure.

  Drug treatment

  Animals were randomly divided into five groups of five animals each (n=5). Group I served as negative control (CMC Na 1% in double distilled water p.o). Group II was given reference standard, Triamcinolon (0.36 mg/kg p.o). Group III-V were administered red ginger extract (GE) 280 mg/kg bw, A. indica extract (AE) 200 mg/kg, and combination of GE-AE 140mg: 100 mg/kg bw, respectively. The pre- pared extract was administered once daily for 7 consecutive days.

  Freund’s adjuvant induced arthritis

  Arthritis was induced by injecting a 0.2 ml of CFA into the left hind paw. Drug treat- ment was started from 5th day after adjuvant injection and continued till 7 days. Paw vol- ume was measured daily from first day after adjuvant induction using plethysmometer. The mean changes in injected paw edema with re- spect to initial paw volume, were calculated on respective days and % inhibition of paw edema with respect to untreated group was calculated (Bendele, 2001).

  RA is a chronic inflammatory disease characterized by fibroblastic proliferation, in- filtration of the synovial lining by inflammato- ry cells which leads to expression of proinflam- matory cytokines and a paucity of apoptosis resulting in bone and joint destruction (Ghildi- yal et al

  ., 2013). Despite widely research being carried out for immune disease, it still remains a challenge since no satisfactory treatment is available in clinical practices.

  Hence, there is enormous interest worldwide for the use of an alternative medicine. The re- search has been focused on formulations used in traditional medicine for treatment of RA (Long et al ., 2001; Ram et al., 2012).

  CFA is used to initiate induction of arthri- tis. This model is the original model of RA, has been extensively used to preclinical screening of new antiarthritis compounds. After a single injection of the adjuvant, a rapid, reliable, ro- bust, and easily measurable polyarthritis de- velops. The joint pathology seen in this animal model shares the cartilage degradation, bone reportion, and cellular influx seen in human RA (Bendele et al., 1999). CFA-induced arthrit- ic animals produced increase in paw volume and paw thickness in disease control group. Treatment with A. indica extracts and triamci- nolone to diseased animal showed statistically significant (P<0.05) reduction in oedema in- hibitory activity compared to disease control group. Extract of red ginger rhizome produced lower effect than that’s of triamcinolone (Fig- ure 1). Based on area under curve (AUC) data of oedema inhibitory activity, AE and combina- tion of GE-AE showed comparable antiarthritis activity to triamcinolone, however GE showed the less AUC (Figure 2).

  ANTIARTHRITIS ACTIVITY OF COMBINATION OF RED GINGER (Zingiber officinale Rosc.) AND ACALYPHA (Acalypha indica Linn)

EXTRACTS ON ARTHTRITIS RAT

  Aktivitas Antiarthritis Kombinasi Ekstak Jahe Merah (Zingiber officinale Rosc.) dan Acalypha (Acalypha indica Linn) Terhadap Tikus arthritis

RESULTS AND DISCUSSION

  Rina Herowati, Gunawan P. Widodo and Ismi Puspitasari

Figure 1. Curve of oedema inhibitory percentage vs days of treatment

  Figure 2. AUC of oedema inhibitory percentage

  Normat rat’s ankle showed clear and wide joint space, but arthritis rat’s ankle showed narrow joint space. Histopathology of ankle joint in CFA treated rats, revealed en- hanced neutrophil infiltration, whereas in tri- amcinolone as well as GE treated rats there were significant reduction in neutrophil infil- tration (Figure 3).

  ANTIARTHRITIS ACTIVITY OF COMBINATION OF RED GINGER (Zingiber officinale Rosc.) AND ACALYPHA (Acalypha indica Linn)

EXTRACTS ON ARTHTRITIS RAT

  Aktivitas Antiarthritis Kombinasi Ekstak Jahe Merah (Zingiber officinale Rosc.) dan Acalypha (Acalypha indica Linn) Terhadap Tikus arthritis

Figure 3. Histopathology of rat’s ankle

  Table 1 presented the average length of joint space on histopathology study. On nor- mal rat group, the joint space length was wide, contrary to the narrow joint space on arthritis group. Therapy with triamcinolone for 7 days significanly increased the length of joint space, and comparable effect was showed by AE group. The effect of AE group was much better than that’s of GE and combination of GE-AE. It was in accordance with the reasearch of Krish- na et al. (2011), which reported the widen of joint space length after AE administration, due to decreasing of capillary permeability. Both of GE and the combination of GE-AE showed less of joint space length, but the joint space was clear from infiltration.

  

Table 2. Length of joint space on histopathology study

Group Length of joint space (μm) GE 280mg/kg

  121,56 ± 56,56 AE 200mg/kg 173,76 ± 63,17 GE:AE 140mg:100mg/kg 111,1 ± 26,02

  Triamcinolone 0.36mg/kg 179,33 ± 64,10 Negative control 104,13 ± 17,06 Normal rat 229,63 ± 35,61

  Rehman et al. (2011) reviewed that ginger constituents inhibit arachadonic acid metabolism and thus prostaglandin synthesis. This may play a role in its anti-inflammatory properties. Ginger may be a stronger inhibitor of prostaglandin synthesis than indometha- cin. It can be used either for rheumatoid or osteoarthritis (Shen et al., 2003). Our phy- tochemical investigation showed the presence of alkaloids, flavonoids, terpenes, and saponin in the Acalypha extract, that could be sug- gested to contribute to the antiarthritis activ- ity of the extract. Each of ginger extract and Acalypha extract showed the preference as antiarthritis activity. Acalypha extract showed oedema inhibitory activity and enhancement of joint space length comparable to triamcino- lone, while ginger extract produced clear joint space. Hence the combination of Acalypha ex- tract and ginger extract has an advantage com- pared to each single extract.

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  CONCLUSION

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  This study was supported by a research grant from DIRJEN DIKTI-Indonesian National Ministry of Education (Hibah Bersaing 2013) for the authors and greatly acknowledged.

  ACKNOWLEDGMENT

  Oral dosage of acalypha extract, red ginger extract, as well as their combination reduced hind paw swelling, comparable to that’s of triamcinolone. Histopathology study revealed the significant reduction in mononu- clear infiltration and increased the ankle joint space. The combination of red ginger rhizome and acalypha extracts showed protective effect against adjuvant induced arthritis.

  Rina Herowati, Gunawan P. Widodo and Ismi Puspitasari

  ANTIARTHRITIS ACTIVITY OF COMBINATION OF RED GINGER (Zingiber officinale Rosc.) AND ACALYPHA (Acalypha indica Linn)

EXTRACTS ON ARTHTRITIS RAT

  Aktivitas Antiarthritis Kombinasi Ekstak Jahe Merah (Zingiber officinale Rosc.) dan Acalypha (Acalypha indica Linn) Terhadap Tikus arthritis

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  Effects of ginger (Zingiber officinale Rosc.) on decreasing the production of inflammatory mediators in sow os- teoarthrotic cartilage explants. J. Med. Food, 6: 323-328.