Anti-Arthritic Activity of Leaf of Carissa carandas (L) against Adjuvant- Induced Arthritis in Rat

Introduction

Carissa carandas is a species of flowering shrub in the dogbane family, Apocynaceae. It produces berry- sized fruits that are commonly used as a condiment or additive to Indian pickles and spices. It is a hardy, drought-tolerant plant that thrives well in a wide range of soils. Common names include Koromcha in Bangla and Karanda in English. Its botanical name was in recent years changed to Carissa congesta Wight (syn. C. carandas Auct formerly widely shown as C. carandas L.) [1]. Phytochemical screening of the root extract showed that the crude extract contained small quantities of alkaloids, flavonoids, Saponins and large amounts of cardiac glycosides, triterpenoids, phenolic compounds and tannins [2]. Earlier studies have shown that the extract of the plant possesses cardiotonic, antipyretic and antiviral activity [3, 4, 5]. Various cardiac glycosides, a triterpenoidal constituent carissone and β-sitosterol were reported from the root extract of the plant [6]. In Western Ghats region of India, the decoctions and extracts of the roots of this plant are effective remedies in the management and/or control of convulsions and epilepsy. However studies showed that root extract of C. carandas may produce its anticonvulsant effects via non-specific mechanisms.

Rheumatoid arthritis is a chronic inflammatory disease that affects the synovial lining between the joints and can lead to severe pain and loss of joint function. RA is a chronic autoimmune disease characterized by poly- articular inflammation associated with synovitis, otitis, and peri-articular osteopenia, often associated with erosion of subchondral bone and progressive joint space narrowing [7]. It is widely acknowledged the early diagnosis of rheumatoid arthritis (RA) and aggressive treatment to control disease activity offer the highest likelihood of preserving function and preventing disability. These features are commonly leaded to progressive joint damage, impaired function, and disability [8, 10].

Since half of the patients of the RA roughly suffer from disability within 10 years of diagnosis, it is critical to effectively treat the disease early to reduce inflammation and prevent damage of bone and joint cartilage [11, 12]. Treatment is commonly determined by the extent or severity of disease activity, and it is assessed by counting the number of swollen and tender joints measuring patient-reported outcomes (for example, patient global quality of life assessment), and assaying the acute phase responses, such as the erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels.

Statistical Analysis

The experimental result is represented as mean ±SEM. The data were statistical analyzed by one way analysis and variance (ANOVA) fallowed by Dunnet’s comparison test. P value< 0.5 was considered as significant.

Result and Discussion

The percentage yield of the ethanolic extract were found to be 24% the ethanolic extract was found to contain alkaloids, glycosides, flavonoids, steroids, terpenoids and carbohydrates, tannins. From the acute toxicity study it was found that the extract were safe up to 4000mg/kg body weight, so one tenth of this dose ( i,e 400mg/kg ) was considered as high dose where as one twenty of this dose ( i,e 200mg/kg ) of body weight was considered as low dose. In the present investigation by standard drug Diclofenac sodium at a dose of 10mg/kg of body weight the paw volume was significantly decreased (p<0.1) from 3 day after the induction of CFA, whereas the extract significantly decreases the paw volume after the 3 day of shown in (Table 2 & Figure 1).

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In the Table 3 it was found that ethanol extract has got highest percentage inhibition 62.66% in female rat at a dose of 400mg/kg of body weight when compared with the low dose 200mg/kg which is 54.67%.at the end of 21th days. Standard drug Diclofenac decreases the paw volume 53.09% [15]. The radiographic images of the knee joints of all groups of rats are shown in Figure 2. It is evident from the radiographic images that adjuvant treated rats developed joint destruction, sclerosis and osteophytes and soft tissue swelling was seen. Whereas, in the extracts treated animals at a dose (400mg/kg) of body weight and standard Diclofenac at a dose of (10mg/kg) of body weight showed shows mild inflammation and no osteophytes were seen when compared to control group. In the present study, we also examined joint histopathology. The affected knee joint of each animal were removed at the end of experiment and processed by radiographs and H and E staining. The radiographs showed a significant loss of bone, periosteal bone formation and also caused soft tissues swelling. Whereas as groups treated with extract of leaf of Carissa carandas at a dose of (400mg/kg) of body weight and treated standard drug Diclofenac sodium at a dose of (10mg/kg) of body weight showed good protection against the CFA induced arthritis in rats. The extract treated group’s shoed lowered radiographic scores when compared negative control. The images of H and E staining section of arthritic group showed disruption of normal architecture of synovial membrane with severe infiltration of lymphocytes and invasion of inflamed synovial membrane with the articular bone and loss of articular cartilage [16].

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The haematological data revealed that there was an increase in RBC count and haemoglobin level, and decrease in the ESR. Further, there was a significant decrease in leukocyte count in extract treated groups while it was reverse in control group shown in Table 4 These data suggest that Carissa carandas ethanolic leaf extract has significant role in the arthritis [17] (Figures 3- 6).

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