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| | | Year : 2009 | Volume : 3 | Issue : 4 | Page : 281-284 | | | Comparing the anti-arthritic activities of the plants Justicia gendarussa Burm F. and Withania somnifera Linn | | | Jaijesh Paval1, Srinivasan Keloth Kaitheri2, Bhagath Kumar Potu3, Sreejith Govindan4, Raju Suresh Kumar5, Sareesh Naduvil Narayanan5, Sudheer Moorkoth2
1 Department of Anatomy, Melaka Manipal Medical College, Manipal, Karnataka, India
2 Department of Pharmaceutical Chemistry, Manipal College of Pharmaceutical Science, Manipal, Karnataka, India
3 Department of Anatomy, Kasturba Medical College, Manipal, Karnataka, India
4 Department of Microbiology, Melaka Manipal Medical College, Manipal, Karnataka, India
5 Department of Physiology, Melaka Manipal Medical College, Manipal, Karnataka, India
Click here for correspondence address and email | Date of Submission | 21-Mar-2008 | | Date of Acceptance | 31-Dec-2008 | | Date of Web Publication | 6-Feb-2010 | | |      | |  Abstract | | | The aim of this study was to compare the anti-arthritic activities of the plants Justicia gendarussa and Withania somnifera. Arthritis is induced in male albino rats using Freund's complete adjuvant and bovine type II collagen. Leaves of J. gendarussa and roots of W. somnifera were powdered and extracted with ethanol (95%) using the soxhlet method. The effect of these plant extracts on arthritic rats were assessed by various blood parameters and also by taking the change in paw volume. The plants J. gendarussa and W. somnifera suppressed the anti-arthritic changes induced in rats and the results were statistically significant. Keywords: Arthritis, collagen, paw volume, plant extract How to cite this article:
Paval J, Kaitheri SK, Potu BK, Govindan S, Kumar RS, Narayanan SN, Moorkoth S. Comparing the anti-arthritic activities of the plants Justicia gendarussa Burm F. and Withania somnifera Linn. Int J Green Pharm 2009;3:281-4 |
How to cite this URL:
Paval J, Kaitheri SK, Potu BK, Govindan S, Kumar RS, Narayanan SN, Moorkoth S. Comparing the anti-arthritic activities of the plants Justicia gendarussa Burm F. and Withania somnifera Linn. Int J Green Pharm [serial online] 2009 [cited 2014 Mar 7];3:281-4. Available from: http://www.greenpharmacy.info/text.asp?2009/3/4/281/59732 |
| Introduction | |  |
Withania somnifera (Family: Solanaceae), commonly known as ashwagandha, is a medicinal plant widely found in India and North America. [1] The root of this plant contains flavonoids and various compounds of the withanolide group. Pharmacological studies on animal models support the antitumour and radio- sensitizing effects of this plant. [2] The alkaloid fraction of the root extract exhibits hypotensive, bradicardiac and respiratory stimulant activities in dogs. Studies also provide information about the antibacterial, antitumour, anti-arthritic and immunosuppressive properties of this plant. [3]
Justicia gendarussa (Family: Acanthaceae) is a shade loving, quick growing, ever green pant which is mostly found in moist areas. It is considered to be a native of China and is distributed widely in India, Sri Lanka and Malaya. In the Indian and Chinese traditional medicines, the leaves of the plant is recommended to treat ailments such as fever, hemiplegia, rheumatism, arthritis, headache, ear ache, muscle pain, respiratory disorders and digestive troubles. [4],[5]
Although the anti-arthritic effect of W. somnifera is well known, there are no published scientific studies on the anti-arthritic activities of the leaves of J. gendarussa. The objective of this study was to compare the anti-arthritic potential of these two plants using two arthritic rat models.
| Materials and Methods | | |
Plant Material
Leaves of J. gendarussa and the roots of W. somnifera were collected from Udupi (Karnataka district, India) in the month of August 2006. Two kilograms of dried leaves of J. gendarussa and 2 kg of dried roots of W. somnifera were blended to fine powder separately and extracted with ethanol (95%) using the soxhlet method. The extracts were concentrated by distillation under reduced pressure and evaporated to dryness. The total yield of the J. gendarussa extract was 65 g and that of the W. somnifera extract was 20.97 g.
Test Animals
Male albino rats of Wistar strain weighing around 180-200 g were used for the studies. The animals were housed in cages under standard laboratory conditions (12:12 h light/dark cycle at 25 + 5°C). The rats were fed with commercial rat diet and water ad libitum and were divided into groups of six. The ethical guidelines for the investigation of the animals used in experiments were followed in all the tests.
Acute Toxicity Test
A group of six rats was given graded doses of 0.25, 0.5, 1 and 2 g of plant extracts. The rats were continuously observed for their mortality and behavioural responses for 48 h and thereafter once daily until the 14 th day. The selection of a dose is done by taking 1/10 of the lethal dose. Ld50 obtained from J. gendarussa extract was 1000 mg/ kg rat and that from W. somnifera extract also was 1000 mg/kg rat. The experimental dose of both the extracts was selected as 100 mg/kg rat.
Induction of Arthritis
Arthritis was induced by injecting Freund's complete adjuvant (FCA) and bovine type II collagen. In the first method, 0.5 ml of FCA containing 10 mg of dry heat-killed Mycobacterium butyricum/ml of sterile paraffin oil (Difco Laboratories, Detroit, MI, USA) was injected in to the plantar surface of left hind foot of the animal. [6],[7] In the second method, 0.1 ml of collagen emulsified with incomplete Freund's adjuvant (IFA) was injected to the left hind foot. [8] The hind paw swelling in each animal was examined using a plethysmograph.
Experimental Design
Animals were divided into four groups of six rats each as follows: Group I - normal rats; Group II - arthritic rats; Group III - arthritic rats administered with the root extract of W. somnifera; Group IV - arthritic rats administered with the leaf extract of J. gendarussa. Administration of J. gendarussa and W. somnifera was started on the 20 th day of the induction of arthritis and continued for 20 days. During the 20 days of treatment, the paw volume and the weight of the animals were recorded at regular intervals. At the end of the 20 th day, the animals were killed by cervical dislocation. The serum separated from the blood was collected for further biochemical assays.
Phytochemical Screening
Since the components of the ethanolic root extract of W. somnifera were well known, only the J. gendarussa extract was tested to identify the presence of alkaloids, steroids, flavonoids, etc. using standard procedures.
Biochemical Assays
Hemoglobin content was estimated by the method of Drabkin and Austin. [9] Red blood cell and white blood cell counts were estimated according to the method of Chesbrough and Mc Arthur in an improved Neubauer chamber. [10] Estimation of erythrocyte sedimentation rate was followed by the method of Westergren. [11] For the estimation of copper level, the colorimetric bathocuproine disulfonate method of Zak and Landers was used. [12] C-reactive protein level was estimated using the ELISA kit obtained from Alpha Diagnostics Int., USA.
Statistical Analysis
The results were analyzed using one-way analysis of variance followed by the Bonferroni test. The values are expressed as mean + standard deviation.
| Results | | |
Paw Volume
In arthritic rats, the paw volume reached its maximal size on 20 th day in both the arthritic models. Paw maintained its inflamed condition for the next 20 days in Group II rats. A significant reduction in paw volume was observed in W. somnifera-treated group (group III) and J. gendarussa-treated group (group IV) when compared to group II. The results are illustrated in [Figure 1].
Body Weight
[Figure 2] and [Figure 3] shows the body weight of normal and experimental groups of rats. There is a significant decrease in the body weight of arthritic rats (group II) when compared to normal rats (group I). Administration of J. gendarussa and W. somnifera extracts improved the body weight significantly when compared to group II in both collagen and FCA-induced arthritic rats.
Hematological Parameters
[Table 1] and [Table 2] shows hematological parameters such as Hb, RBC count, WBC count, ESR, serum copper level and C-reactive protein level of normal and experimental groups of rats. A significant decrease in levels of RBC and Hb were observed in arthritic rats (group II) when compared to normal rats (group I). Administration of J. gendarussa and W. somnifera extracts to arthritic rats enhanced the levels of Hb and RBC to near normal levels. The increased levels of WBC, ESR, serum C-reactive protein and serum copper were significantly suppressed in the extract-administered arthritic groups.
| Discussion | | |
FCA-induced arthritis and collagen-induced arthritis are the two models which are extensively used to study the pathogenesis of rheumatoid arthritis for testing therapeutics. [7] There is evidence which indicates that arthritis and certain other connective tissue diseases are caused by hypersensitive mechanisms. Adjuvant-induced arthritis has been widely used as an experimental model of such diseases. [7] Collagen-induced arthritis (CIA) is an experimental model sharing several clinical and pathological features with rheumatoid arthritis (RA). The importance of T cells in the pathogenesis of CIA and RA has been established and various studies have been performed to determine the cytokines and susceptibility factors involved in arthritis development. [13],[14]
Paw swelling is one of the major factors in assessing the degree of inflammation and therapeutic efficacy of the drugs. [15] Here, the J. gendarussa-treated rats showed 48% paw oedema inhibition in both FCA-induced arthritic model and collagen-induced arthritic models. W. somnifera- treated rats showed 45% of paw oedema inhibition in the FCA- induced arthritic model and 43% inhibition in the collagen-induced arthritic model.
In the present study, the arthritic rats (group II) showed a reduced RBC count, reduced Hb level and an increased erythrocyte sedimentation rate. All these indicate the anemic condition and is commonly noted in patients with chronic arthritis. [16] The two most common reasons for anemia in arthritic patients are gastrointestinal blood loss from arthritis medications and bone marrow changes in patients with inflammatory arthritis, which prevent the release of iron for incorporation into RBCs. [17,18] The extract- treated groups showed a significant recovery from anemia.
The increase in WBC count in arthritic rats was significantly suppressed in J. gendarussa- and W. somnifera-treated rats.
C-reactive protein is a member of the class of acute phase reactants as its levels rise dramatically during inflammatory processes. [19] The level of CRP is significantly reduced in plant-treated groups.
Ceruloplasmin is an enzyme synthesized in the liver containing eight atoms of copper in its structure. Free copper ions are powerful catalysts of free radical damage. By binding copper, ceruloplasmin prevents free copper ions from catalyzing oxidative damage. [20] The increased level of copper ion indicates the inflammatory condition. [21] Serum copper concentration was measured in normal and arthritic rats. The arthritic rats exhibited a significant elevation of copper level and this was suppressed in J. gendarussa- and W. somnifera-treated rats.
The phytochemical analysis of the J. gendarussa extract showed the presence of flavonoids vitexin and apigenin. The plant J. gendarussa was reported to yield â-sitosterols, alkaloids, reducing sugars and unidentified sterols. [22] The compounds such as flavonoids and â-sitosterols are well known for their anti-inflammatory activities and the presence of these compounds in the leaf extract may be the reason behind the anti-arthritic properties shown by this plant.
The chemistry of W. somnifera has been widely studied and many chemical components have been isolated. [23] They include alkaloids (isopelletierine, anaferine), steroidal lactones (withanolides, withaferins), saponins containing an additional acyl group (sitoindoside VII and VIII) and withanolides with a glucose at carbon 27 (sitoindoside IX and X). [24]
There have been various studies to support the anti- arthritic activities of the plant W. somnifera. The compound Withaferin A was found to exhibit fairly potent anti-arthritic and anti-inflammatory activities. It was found to suppress the adjuvant-induced arthritic changes in rats without any toxic effect. [25],[26],[27]
| Conclusion | | |
This study clearly showed that the anti-arthritic property of the plant J. gendarussa can be comparable with that of the W. somnifera. Further studies are needed in the isolation of other compounds that may be involved in determining the anti-arthritic potential of the plant J. gendarussa.
| References | | |
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Correspondence Address:
Jaijesh Paval
Department of Anatomy, Melaka Manipal Medical College, Manipal, Karnataka
India
  | 3 |
DOI: 10.4103/0973-8258.59732
[Figure 1], [Figure 2], [Figure 3]
[Table 1], [Table 2] | |
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