EVALUATION OF ANTIARTHRITIC ACTIVITY OF LEPIDIUM SATIVUM LINN SEEDS AGAINST FREUND'S ADJUVANT INDUCED ARTHRITIS IN RATS

Abstract

Garden cress leaves and seeds are used in India as food supplement and also as medicine. In traditional system of medicine, its seeds were used for various ailments like inflammation, joint pain, backache etc the present study was carried out to explore the folkloric use of the seeds on Freund's adjuvant induced arthritic rats. This present study concludes the effect of the seed powder on the Freund's complete adjuvant induced arthritic rat paw oedema in both developing and developed phases of arthritis. Histopathology of proximal inter-phalangeal joints and radiology of hind legs were studied. Significant changes were observed, in drug treated group in radiological study. In histo-pathological study bone and cartilage degeneration with bone erosion and inflammatory changes were observed in the affected joint. These changes were not significantly improved by the administration of test drugs. Result data suggested that, at a lower dose, moderate anti-arthritic effect was produced by the test drug, while at a higher dose it had a tendency to produce inconsistent effect.

Headnote

ABSTRACT

Garden cress (Lepidium sativum Linn.) leaves and seeds are used in India as food supplement and also as medicine. In traditional system of medicine, its seeds were used for various ailments like inflammation, joint pain, backache etc. the present study was carried out to explore the folkloric use of the seeds on Freund's adjuvant induced arthritic rats. The present study concludes the effect of the seed powder on the Freund's complete adjuvant induced arthritic rat paw oedema in both developing and developed phases of arthritis. Histopathology of proximal inter-phalangeal joints and radiology of hind legs were studied. Significant changes were observed in drug treated group in radiological study. In histo-pathological study bone and cartilage degeneration with bone erosion and inflammatory changes were observed in the affected joint. These changes were not significantly improved by the administration of test drugs. Result data suggested that at a lower dose, moderate anti-arthritic effect was produced by the test drug while at a higher dose it had a tendency to produce inconsistent effect.

KEY WORDS: anti arthritic, Fruend's adjuvant, paw oedema, Lepidium sativum Linn.

INTRODUCTION:

Rheumatoid arthritis (RA) is a systemic autoimmune disease of unknown aetiology. The disease is characterized by articular inflammation and by the formation of an inflammatory and invasive tissue, rheumatoid pannus that eventually leads to the destruction of joints (Stephen J. 2008). Analgesics and NSAIDs are used to suppress the symptoms while disease-modifying anti-rheumatic drugs (DMARDs) and newer therapies such as anti-tumour necrosis factor (TNF)-α therapy (etanercept, infliximab and adalimumab), anti-CD20 therapy (rituximab) and abatacept are often required to inhibit or halt the underlying immune process (Anthony S. Fauci. et al., 2008). NSAIDs are widely prescribed all over the world to the patients with Rheumatoid arthritis, Osteoarthritis etc. However the prolonged use of these drugs has its own drawbacks. Research works on NSAIDs suggest that these drugs can increase the risk of chronic renal disease (P Ejaz et al., 2004).

Garden cress is also known as Asalio or Chandrashura in local languages and it is an important medicinal crop in India (Tiwari et al, 2004). Garden cress (Lepidium sativum Linn.) is an annual fast growing edible herb, belongs to the family Brassicaceae. Seeds, leaves and roots of it are of economic importance; however, the crop is mainly cultivated for seeds. It can be grown at all elevations, throughout the year, but the best crop is obtained in the winter season (Anonymous, 1962, CSIR, New Delhi.). Garden cress seeds and leaves are used in food preparations (Datta PK et al., 2011). Leaves are diuretic and gently stimulant (Maghrani et al., 2005). To investigate the folkloric use of the seeds the present study was carried out on Freund's adjuvant induced arthritic rats.

MATERIALS AND METHODS:

Collection of sample (Lepidium sativum Linn seeds)

The seeds for the proposed study were collected from the campus of Gujarat Ayurved University, Jamnagar in the month of April 2004 and it was authenticated by the expert of botany from the Department of Pharmacognosy, Gujarat Ayurved University, Jamnagar, Gujarat, India

Experimental models:

Twenty four Charles Foster strain albino rats weighing 180 ± 10g were obtained from animal house attached to Pharmacology laboratory, Gujarat Ayurved University, Jamnagar Gujarat. Six rats were housed in each cage made up of poly-propylene with stainless steel top grill. The dry wheat (post hulled) waste was used as bedding material and was changed every morning. The animals were acclimatized for seven days before commencement of the experiment in standard laboratory conditions 12 ± 01 hour day and night rhythm, maintained at 25 ± 3°C and 50-70% humidity as per CPCSEA guidelines. Animals were provided with balanced food (Amrut brand rat pellet feed supplied by Pranav Agro Mills Pvt. Limited) and water ad libitum. Protocol used in this study for the use of animals was approved by the institutional animal ethics committee (IAEC 04-05/01/PhD.03).

Dose selection:

The dose fixation for the experimental animals was done on the basis of body surface area ratio by referring to the standard table of (Paget and Barnes, 1964). The adult human dose (6 g per day) (Chunekar K.C, 1999) was converted to animal dose. On this basis the calculated dose was fixed to be 550 mg/kg for rats. The suspension of Lepidium seed powder was made with sufficient quantity of distilled water according to the required dose and administered orally with the help of gastric catheter sleeved to syringe.

Experimental study (Rosenthale, 1970):

The selected animals were grouped into three groups of 6 rats each. To the first group only water was administered and treated as normal control. To second and third group, test drug was administered in the dose of 550 mg/kg (TED) and 1100 mg/kg (TED × 02) respectively. The drug was administered for 30 consecutive days. On day one, the complete Fruend's adjuvant was made into fine emulsion with the help of a syringe and 0.1 ml of it was injected beneath the plantar aponeurosis in the lefthind paw and 0.05 ml subcutaneously into the root of the tail. The volumes of both the hind paws were measured with the help of Plethysmometer. The paw volume of lefthind limb was measured at 2nd, 3rd, 5th, 10th and 15th days, while of right hind limb on 15th, 20th, 25th and 30th days. Paw volume of the 0 (initial) days were taken as the reference value for determining the increase in paw volume on the subsequent days. On 31st day animals were weighed and sacrificed by over dose of ether anesthesia and both right and leftsynovial joints were dissected out and the histo-pathological slides were prepared by referring to standard procedure of (Raghuramulu et al., 1983). The slides were viewed under trinocular research Carl-Zeiss's microscope at various magnifications to note down the changes in the microscopic features.

Statistical analysis:

The data were expressed as mean ± standard error mean (SEM). The Significance of differences among the test drugs and control groups was assessed using one way analysis of variance (ANOVA) and the test followed by Dunnet's test. The difference was considered significant when p < 0.05.

RESULTS AND DISCUSSION:

The data pertaining to the effect of test drug on Friend's adjuvant induced arthritis in rats have been tabulated in Table 1 & Table 2

A moderate decrease in both primary and secondary edema was observed in therapeutic dose, and double dose group in comparison to control group. The suppression of primary edema in therapeutic dose was as follows. After 48 hrs 14%, 5th day 34.32%, 10th day 49.01%, 15th day 45.40%, 20th day 33.21%, 25th day 82.32%, 30th day 33.84%. Whereas marginal increase was observed after 24 hrs. In double dose group the suppression of primary edema was after 24 hrs 15.88%, 48hr 4.96%, 5th day 29.15%, 10th day 32.64%, 15th day 51.82%, 30th day 12.34%. Here the increase was observed on 20th and 25th day.

In secondary edema the suppression observed in therapeutic dose group was as follows on 15th day 40.07%, 20th day 41.21%, 25th day 45.33%, 30th day 40.45%. In double dose group 15th day 6.92%, 25th day 2.86%, 30th day 28.57% suppression of edema was observed where as on 20th day marginal increase was observed in double dose group.

Radiology of hind legs in adjuvant induced arthritic rats:

In arthritic disease radiographic changes are useful diagnostic measures which indicate the severity of the disease. In the early stage softtissue swelling can be seen, whereas severe radiological changes like bony erosion and joint space narrowing can be observed in developed stage of arthritis. In adjuvant induced arthritic rats softtissue swelling and cartilage erosions were observed. The degrees of degenerative changes were less in test drug treated group in comparison to control group. The radiographic features of the rat joints in adjuvant induced arthritic rats are shown in Figure 1.

Effect on histopathology of joints:

Figure 2 shows representative sections of inter-phalangeal joint from different groups. Bone and cartilage degeneration with bone erosion and inflammatory changes were observed in the knee joint. These changes were not significantly improved by the administration of test drugs.

The main aim of the present study was to find out the scientific basis for the reported efficacy of test drug in arthritis. Hence it was evaluated against Fruend's adjuvant induced arthritis in rats. Injection of adjuvant elicits immune response of cell-mediated type and produces an arthritic syndrome. Arthritic rats show swelling of the softtissue around ankle joint during initial phases of arthritis. This is mainly due to edema of periarticular tissues such as ligaments and joint capsules (Sanmugapriya et al., 2010). Like human RA, it is also characterized by synovitis, infiltration of neutrophils and proliferative response leading to synovial fibroplasias and periosteal bone deposition (Hazeena et al., 1980).

In the present study a persistent moderate suppression of primary oedema ranging between 14 to 49% was observed at lower dose up to 20th day of observation. The secondary oedema represents oedema of immunological origin (Yoshikawa et al., 1985). In the secondary oedema a consistent 40 to 45% suppression was observed at lower dose indicating that the test drug at this dose has moderate suppression effect on oedema of immunological origin. At higher dose inconsistent effect was observed. The suppression ranged between 5.74% to 54%.

Radiographic changes in RA conditions are useful diagnostic measures which indicate the severity of the disease. Softtissue swelling is the earlier radiographic sign, whereas prominent radiographic changes like bony erosions and narrowing of joint spaces can be observed only in the developed stages (final stages) of arthritis (Harris ED, 1990). The radiographic features of the rat joints in adjuvant induced arthritic model are shown in plate 2. The degrees of degenerative changes were less in test drug treated group in comparison to control group.

CONCLUSION

At lower dose level moderate anti-arthritic activity was observed in FA rats. Higher dose level produced inconsistent effect. Hence it would be necessary to identify suitable dose. This indicates the complex nature of the seed powder. The study confirms the anti-inflammatory and anti-arthritic activity potential of the plant material however; further refinement in the form in which it is administered along with optimum dose fixation is required for optimum therapeutic application of this plant. It would also be interesting to study the other parts of the plant. It is possible that they may also express significant anti-inflammatory and anti-arthritic activity.

Sidebar

Cite this article:

Raval Nita. D., Ashok. B.K,, Ravishankar. B., (2013) EVALUATION OF ANTIARTHRITIC ACTIVITY OF LEPIDIUM SATIVUM LINN SEEDS AGAINST FREUND'S ADJUVANT INDUCED ARTHRITIS IN RATS, Global J Res. Med. Plants & Indigen. Med., Volume 2(7): 532-537

References

REFERENCES

Anonymous. (1962). The Wealth of India, Raw Materials. VI. Publication and information Directorate, CSIR, New Delhi. India, p.71-73.

Anthony S. Fauci. and Dr. Longo(2008) Harrison's Principles of Internal Medicine, Section 2, 17th edition, part 14, section 2 chapters 314.

Chunekar K.C., (1999) G.S.Pandey (eds) Bhavprakash nighantu, haritakyadi Verga, Chaukhambha orientalia publication.pp 39.

Datta PK, Diwakar BT, Viswanatha S, Murthy KN, Naidu KA, (Mar-April 2011). Safety evaluation studies on Garden cress Lepidium sativum L. seeds in Wistar rats. IJARNP-HS Publications Vol. 4 (1), pp. 37-43.

Dr. Yograj Sharma, (2003-2004). Efficacy of sowing methods, fertilizer application and growth regulations in cultivation of Lepidium sativum Linn. Published thesis (MSc), Gujarat Ayurved University, Jamnagar.

Harris ED (1990). Rheumatoid arthritis. Pathophysiology and implications for therapy. N Eng J Med, 322(18):1277-1289.

Hazeena Begum, V., Sadique, J. (1980). Ind. J.exp. Biol., 26,877.

Maghrani M, Zeggwagh NA, Michel JB. Eddonks M. (2005). Antihypertensive effect of Lepidium sativum L. In spontaneously hypertensive rats. J Ethnopharmacol 100: 193-197.

P Ejaz, K Bhojani, VR Joshi, (2004). NSAIDs and kidney, JAPI, vol.52, pp-632.

Paget, G.E., Barnes, J.M. (1969). In Evaluation of drug activity; Pharmacometrics (Laurence, D.R., Bacharacha, A.L.) Vol. 1, Academic Press, New York

Raghuramulu N, Nair KM and Kalyanasundaram S (1983). A manual of laboratory techniques, (National Institute of Nutrition, Hyderabad).pp. 246-53.

Rosenthale, M.E. (1970). Arch. Int. Pharmacodyn. 188. In: Dahanukar, S., Sharma, S., Karandikar, S.M.(1983). Indian Drugs, 20 (10), 405.

Sanmugapriya Ekambram, Senthamil Selvan Perumal, and Venkataraman Subramanian, (2010). Evaluation of antiarthritic activity of Strychnos potatorum Linn seeds in Freund's adjuvant induced arthritic rat model.BMC, Complementry and Alternative medicine, vol.10.

Stephen J. Mc Phee, (2008). Current Medical diagnosis & treatment, Maxine papadakis publication. Mc Graw hill Lange Chapter Musculo skeletal disorder.

Tiwari PN, Kulmi GS. (2004). Performance of Chandrasur (Lepidium sativum) under different levels of nitrogen and phosphorus. J Med Arom Pl Sci 26: 479-481.

Yoshikawa T, Tanaka H, Kondo M (1985). The increase in lipid peroxidation in rat adjuvant arthritis and its inhibition by Superoxide dismutase. Biochem. Med, 33:320-326.

Source of Support: Nil Conflict of Interest: None Declared