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The petroleum ether fraction of Celastrus paniculatus Willd. seeds demonstrates anti-arthritic effect in adjuvant-induced arthritis in rats


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The petroleum ether fraction of


paniculatus Willd. seeds demonstrates

anti-arthritic effect in adjuvant-induced arthritis

in rats

Pankaj S. Kothavade, Vipin D. Bulani, Padmini S. Deshpande,

Amrita S. Chowdhury, Archana R. Juvekar


Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Matunga, Mumbai, 400 019, India

Received 13 January 2015; accepted 13 May 2015

Available online 31 March 2016

KEYWORDS Celastrus paniculatus; Rheumatoid arthritis; Freund’s complete adjuvant; Cytokine

Abstract Objective: Celastrus paniculatus (CP) Willd. is a plant indigenous to India with me-dicinal properties. It is used in the ayurvedic treatment of inflammatory ailments such as rheu-matoid arthritis. The present study was designed to investigate the therapeutic efficacy of the petroleum ether fraction (PCP) obtained from CP seeds on adjuvant-induced arthritis in rats. Methods: Arthritis was induced in SpragueeDawley rats by immunization with Freund’s com-plete adjuvant (FCA). Arthritis severity was evaluated by arthritis score, paw volume, tibiotar-sal joint thickness, body weight, dortibiotar-sal flexion pain, motility test, stair climbing ability and index of thymus and spleen. Moreover, histopathology of knee joints supported by haematolo-gical analysis was used to assess the anti-arthritic action of PCP. The levels of superoxide dis-mutase (SOD), glutathione (GSH), catalase (CAT), nitric oxide (NO) and malondialdehyde (MDA) in liver were also assessed. Serum samples were collected for estimation of aspartate transam-inase (AST), alanine transamtransam-inase (ALT) and alkaline phosphatase (ALP). In addition, cytokines such as tumour necrosis factor-alpha (TNF-a) and interleukin-6 (IL-6) were estimated in plasma.

Results: PCP significantly alleviated arthritic progression with regards to paw swelling, arthritic score, immune organ indices, hyperalgesic effect and body weight. This phenomenon was correlated with significant suppression of overproduction of inflammatory cytokines (TNF-a (TNF-and IL-6), oxid(TNF-ant stress m(TNF-arkers (MDA (TNF-and NO) (TNF-and cellul(TNF-ar enzyme (AST, ALT (TNF-and ALP) levels versus arthritic rats without treatment. Moreover, PCP restored the decreased levels of SOD, CAT and GSH.

* Corresponding author. Tel.:þ91 022 3361 2215; fax: þ91 022 3361 1020. E-mail address:juvekar.archana@gmail.com(A.R. Juvekar).

Peer review under responsibility of Beijing University of Chinese Medicine.


2095-7548/ª 2015 Beijing University of Chinese Medicine. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

H O S T E D BY Available online atwww.sciencedirect.com



Conclusion: Our results suggest that the anti-arthritic properties of PCP may be due to immunosuppressive effects, cytokine regulation, antioxidant effects and bone-protective activities.

ª 2015 Beijing University of Chinese Medicine. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/ by-nc-nd/4.0/).


Rheumatoid arthritis (RA) is a chronic autoimmune dis-ease, mainly affecting the joints and characterised by synovial hyperplasia, inflammatory cell infiltration and progressive destruction of cartilage and bone.1 Pro-inflammatory mediators such as cytokines, growth fac-tors and antibodies, including RA factor, are considered to be involved in the pathogenesis of this condition. More-over, lipid inflammatory mediators, in particular prosta-glandin E2 (PGE2), contribute to vasodilation, vascular permeability, pain, cytokine and proteinase production in inflamed tissue.2 Therapeutic effects in rheumatic dis-eases might be achieved by antagonizing these pro-inflammatory mediators. Newer therapies such as anti-tumour necrosis factor (TNF)-a therapy (etanercept, infliximab and adalimumab), anti-CD20 therapy (ritux-imab) and CD80/86 blockade (abatacept) are required to inhibit the underlying immune process, while analgesics and anti-inflammatory drugs are used to suppress the symptoms.3 However, all these anti-rheumatic drugs in clinical use are associated with numerous side effects coupled with lengthy treatment duration. Consequently, natural herbal therapies are considered to be safe and effective in suppressing the chronic pain associated with the disease.4

Celastrus paniculatus (CP) Willd. (Celastraceae), known as ‘Black Oil plant’, is a native species distributed throughout Southeast Asia, India and Taiwan.5 CP seeds are commercially available in the medicinal flora market and are used in folk medicine for the treatment of anal-gesia, inflammation and rheumatic pain.6 The plant ex-hibits a varying range of therapeutic effects associated with conditions such as cognitive dysfunction, epilepsy, insomnia, gout, rheumatism and dyspepsia.7 Previous phytochemical studies have reported that CP is composed of a mixture of different compounds, such as palmitic, stearic, oleic, linoleic and linolenic acids, and sesquiter-pene alkaloids, namely celapanin, celapanigin and cela-pagin, sesquiterpene polyol ester, and phytosterols includingb-sitosterol, campesterol and stigmasterol.7,8As part of our ongoing research, we have recently reported the anti-inflammatory activity of the petroleum ether fraction (PCP) and methanolic extract of CP in rats.9 Bioassay-guided fractionation studies revealed that the anti-inflammatory effects of the PCP fraction were medi-ated by suppression of pro-inflammatory cytokines and PGE2. Herein, we aim to investigate the therapeutic effect of PCP on adjuvant-induced arthritis in experimental rats and its underlying mechanisms.

Material and methods

Plant materials

Dried seeds of CP were purchased from a commercial herb market of Mumbai, Maharashtra, India. The plant material was authenticated by Prof. Ganesh Iyer, Ramnarain Ruia College, Matunga (East), Mumbai. A voucher specimen (2012/02) of the plant was deposited at the herbarium of the Department of Pharmaceutical Science and Technology, Institute of Chemical Technology, Mumbai, India.


Freund’s complete adjuvant (FCA) (containing 1 mg of dry heat-killed Mycobacterium tuberculosis per 1.0 mL sterile, paraffin oils) and reference drug indomethacin were pur-chased from SigmaeAldrich (St Louis, MO, USA).

Fraction preparation

The dried seeds of CP (2.5 kg) were powdered and extrac-ted with methanol using Soxhlet apparatus. The filtrates were evaporated by rotary evaporator and 618 g of total extract was obtained. The methanolic crude extract of CP was fractionated by sequential solvent partition using pe-troleum ether, chloroform, ethyl acetate and n-butanol, successively. For this study, the petroleum ether fraction was chosen for further evaluation of anti-arthritic activity on the basis of its promising antioxidant and anti-inflammatory properties.


SpragueeDawley rats (180e200 g) were obtained from the Animals house, Haffkine Bio-Pharmaceutical (Mumbai, India). The animals were maintained in a 12/12 h light/ dark cycle, at controlled room temperature (25 1C) and relative humidity (55 5%). The animals were fed a rodent standard diet with free access to water ad libitum. All the animals were acclimatised to the laboratory environment for 1 week before the experiment. The Animal Ethics Committees of the Department of Pharmaceutical Sci-ences and Technology, Institute of Chemical Technology, Mumbai approved all experimental protocols in accor-dance with Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) guidelines.


Adjuvant-induced arthritis in rats

Arthritis was induced in rats according to the method of Newbould 1963.10Briefly, 30 min after administration of the vehicle/drug, 0.1 mL of FCA was injected intraplantar into the right hind paw of rats, while an equal amount of saline was injected into the left paw on Day 0. Treated rats received PCP (100, 200 and 400 mg/kg) or indomethacin (1 mg/kg) for 28 days. Negative control animals were given 0.5% sodium carboxymethyl cellulose.

Evaluation of arthritis

Measurement of paw volume, tibiotarsal joint thickness and body weight

Paw volume, tibiotarsal joint thickness and weight of the injected animals were measured on Days 0, 7, 14, 21 and 28 days after vehicle/drug administration.11

Assessment of arthritic score

The severity of arthritis was assessed on a scale of 0e4 with the following criteria: 0 Z no oedema or swelling, 1 Z slight oedema and limited erythema, 2 Z slight oedema and erythema from the ankle to the tarsal bone, 3Z moderate oedema and erythema from the ankle to the tarsal bone, and 4Z oedema and erythema from the ankle to the entire leg. The sum of the scores for all four limbs was calculated as the arthritic index, with a maximum possible score of 16 per rat.12

Dorsal flexion pain13

Dorsal flexion pain (DFP) of the injected paw was scored on the basis of squeaking and leg withdrawal after the ankle joint of the rat was flexed five times until the toes touched the front of the leg. Score 0, no squeaking and leg with-drawal; 1, either squeaking or leg withwith-drawal; 2, both squeaking and leg withdrawal.

Motility test and stair climbing ability13

Motility was observed for a period of 5 min and scored from 0 to 2 points. Score 0: rats avoided touching the feet to the ground; 1, walked with difficulty with toes touching the ground; 2, walked easily. In the stair climbing ability test, rats were allowed to climb on a staircase with steps at 5, 10 and 15 cm, with access to water at the second step and food at the third step. This stair climbing ability was scored from 0 to 3: Score 0 if the rats did not climb any step; 1, climbed onto Step 1; 2, climbed onto Step 2; and 3, climbed onto Step 3. Thymus and spleen index

At the end of the experiment, the animals were sacrificed after blood sampling. The thymus and spleen were imme-diately removed from each animal and weighed. The indices of the thymus and spleen were expressed as the ratio (mg/g) of the thymus and spleen wet weight versus body weight, respectively.11

Haematological analyses

On Day 28, blood samples were collected in EDTA-containing tubes by retro-orbital vein puncture and the total and differential leucocyte counts were determined.

Liver biochemical analysis

Animals were sacrificed on Day 28 post-inoculation; the liver was promptly removed, stored at 70C and later homogenized. The homogenate was centrifuged at 6000g at 4C for 20 min and the supernatant collected. This su-pernatant was used to measure superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), malon-dialdehyde (MDA) and nitric oxide (NO) activity.11

Plasma and serum analysis

The levels of alkaline phosphatase (ALP), aspartate trans-aminase (AST), and alanine transtrans-aminase (ALT) in serum were determined using quantitative colourimetric assay kits, according to the manufacturer’s instructions. Interleukin-1 (IL-1) and TNF-a in plasma were determined using commercially available rat cytokine ELISA kits (Krishgen BioSystems, India), according to the manufac-turer’s instructions.11

Histopathological evaluation of knee joints

On Day 28 after FCA injection, all rats were sacrificed following blood collection. The knee joint of the injected paw was removed for histopathological examination. The joints were fixed in 10% phosphate buffer formalin, decal-cified in 10% EDTA for 30 days at 4C and then embedded in paraffin wax. Sections of 5mm were stained with haema-toxylin and eosin (H&E) and examined under light microscope.

Statistical analysis

Data were expressed as the mean (SEM), and statistical comparisons were carried out using one-way analysis of variance (ANOVA) followed by a post-hoc Dunnett’s test for comparison between control and test groups. The DFP, stair climbing ability test and motility scores were expressed as median scores and the KruskaleWallis test was used to compare the groups. The minimal level of significance was identified at P< .05.


Rat hind paw volume

The effect of the PCP fraction on inflammation in FCA-induced arthritis was examined by measuring the changes in paw volume. As shown inFig. 1a, progressive swelling of injected hind paws increased in arthritic rats over time until the experimental endpoint at Day 28. Treatment groups receiving 200 or 400 mg/kg PCP, or the standard drug indomethacin demonstrated significantly reduced paw oedema volume when compared with the untreated arthritic group (P< .001).

Tibiotarsal joint thickness

As shown inFig. 1b, the thickness of tibiotarsal joints of arthritic rats did not change significantly during the first week but increased from Day 7 to Day 28. In the untreated arthritic group, changes in tibiotarsal thickness continued


to increase progressively, reaching significantly higher values on Day 28. Treatment with 200 or 400 mg/kg of the PCP fraction significantly decreased the tibiotarsal joint thickness on Days 14, 21 and 28 compared with untreated arthritic rats (P< .001), as did indomethacin treatment.

Body weight

Weight gain in the untreated FCA control group was significantly impaired throughout the experiment compared with non-arthritic rats (Fig. 2). After treatment with PCP

a b *** *** *** *** * * *** *** *** *** 0.900 1.200 1.500 1.800 2.100

0 days 7 days 14 days 21 days 28 days

Paw Volum e (m l) FCA Control Indomethacin PCP 100 mg/kg PCP 200 mg/kg PCP 400 mg/kg *** *** *** ** *** *** *** *** *** *** 0.00 5.00 10.00 15.00 20.00 25.00 30.00 35.00

7 days 14 days 21 days 28 days

% changes in tibiotrsal thick ness FCA Control Indomethacin PCP 100 mg/kg PCP 200 mg/kg PCP 400 mg/kg

Figure 1 Effect of PCP on (a) paw swelling and (b) % changes in tibiotarsal thickness. All data are expressed as the mean (SEM; nZ 6). *P < .05, **P < .01 and ***P < .001 compared with the FCA control group.

Figure 2 Effect of PCP on body weight in arthritic rats. Values are expressed as means (SEM), nZ 6. *P < .05, **P < .01 and ***P< .001 compared with the FCA control group.


(200 and 400 mg/kg) or indomethacin, the arthritic rats exhibited dose-dependent body weight gain. However, no remarkable body weight increase was observed in the PCP 100 mg/kg group.

Arthritic score

The severity of arthritis was measured on Days 7, 14, 21 and 28 and scored by physical observation. Groups treated with 200 and 400 mg/kg PCP significantly suppressed swelling in a dose-dependent manner, thereby reducing the arthritic score in the FCA-injected paw (P< .01). Treatment with indomethacin was also effective at limiting arthritic score in the affected paw. However, 100 mg/kg PCP had no sig-nificant effect (P> .05;Fig. 3).

Dorsal flexion pain

The FCA control group rats exhibited a low threshold of pain and a median score of 2 was obtained on Day 28. Treatment with indomethacin and 400 mg/kg PCP significantly ameliorated the pain, with animals in these treatment groups demonstrating a median score of 1 on Day 28 (P< .05;Fig. 4a).

Motility test

As shown in Fig. 4b, FCA-induced arthritic rats showed impairment in motility from Day 14 after injection of FCA and a median score of 0 was obtained. Treatment with 200 mg/kg and 400 mg/kg PCP ameliorated motility impairment in a dose-dependent manner, with median scores of 1 and 2, respectively, being obtained on Day 28 (P< .05).

Stair climbing ability

Injection of FCA into rat paws affected the animals’ stair climbing ability and a median score of 0.5 was obtained in the FCA control group on Day 28, compared with a median score of 3 in the non-arthritic control group (Fig. 4c). Treatment with 400 mg/kg PCP resulted in an improved median score from 1.5 to 3 on Day 7 and Day 28, respectively.

Thymus and spleen index

Our results indicated that the thymus and spleen index of the FCA control group increased compared with the non-arthritic control group. However, treatment with PCP demonstrated a marked dose-dependent reduction of these measurements (Table 1).

Haematological examinations

Changes in peripheral blood profile were observed on Day 28 post injection of FCA into the hind paw of rats. There were no significance differences in the haematological profiles of PCP-treated rats compared with the FCA control group (Table 2).

Liver biochemical examination

A significant (P< .05) decrease in the levels of SOD, CAT and GSH was observed in arthritic rats compared with normal controls (Fig. 5). The administration of PCP (200 and 400 mg/kg) and indomethacin (1 mg/kg) significantly attenuated the reduction of these enzymes when compared with the FCA control group (P< .05). Meanwhile, MDA and NO levels were significantly higher in arthritic rats than normal controls (P < .05). Treatment with PCP and indo-methacin reduced the levels of MDA and NO. Overall, all antioxidant indices tested were modulated significantly and PCP treatment exhibited a satisfactory antioxidant effect. Biochemical analysis in serum

Significant elevation of AST, ALT and ALP was observed in the serum of adjuvant-induced arthritic rats compared with normal healthy animals (P< .001;Fig. 6). Administration of indomethacin (1 mg/kg) and the higher doses of PCP significantly decreased the levels of AST, ALT and ALP compared with FCA control rats (P< .05).

Cytokine levels in plasma

As shown inFig. 7, the levels of cytokines in plasma were significantly elevated in arthritic rats compared with

Figure 3 Effect of PCP treatment on arthritis score in arthritic rats. Values are expressed as means (SEM), nZ 6. **P < .01 and ***P< .001 compared with the FCA control group.


Figure 4 Effect of PCP on inflammatory hyperalgesia in arthritic rats. The scores of (a) dorsal flexion pain, (b) motility and (c) stair climbing ability are illustrated as box plots where a bold line represents median values (nZ 6); boxes represent interquartile ranges (25th and 75th percentiles) compared with the FCA control group. *P< .05 and **P < .01.


normal controls (P< .001). In all drug-treated groups, rats exhibited significant reductions in TNF-a and IL-6 levels compared with arthritic controls (P< .001).

Histopathological examination

The histopathological examination of tibiotarsal joints of untreated FCA-induced arthritic rats showed marked syno-vial hyperplasia, inflammatory cell infiltration and cartilage or bone destruction. The arthritic rats administered with 200 or 400 mg/kg PCP, or 1 mg/kg indomethacin showed a remarkable reduction in synovial hyperplasia and inflam-matory cell infiltration compared with arthritic controls (Fig. 8).


Arthritis is an inflammatory condition that is associated with joint pain, deformity and restricted mobility. The retention of microbial products in the synovial tissue and persistent infection of the joint articular surface induce an immune reaction, ultimately altering the integrity of these components of the joint.3In view of the anti-inflammatory properties of the C. paniculatus and its bioactive fraction PCP,9 the present study was carried out with the aim of investigating efficacy against the symptoms of arthritis, including physical changes and inflammation. Adjuvant-induced arthritis in rats is widely used to study the

pathophysiology of RA as an experimental model demon-strating various features of rheumatoid arthritis observed in humans.14 Intra-articular injection of FCA-induced an in-flammatory response characterised by the release of TNF-a and IL-6, and inflammatory mediators including vasoactive amines (histamine and serotonin) and prostaglandins.13,15 Significant increases in paw thickness after FCA sub-plantar administration reflected the onset of arthritis. We found that treatment with 200 and 400 mg/kg of PCP significantly decreased paw swelling and thickness by inhibiting the release of such inflammatory mediators.

In rats, body weight is directly proportional to food intake and metabolism, but this might be affected by im-munity and inflammation, regulated by the hormone leptin.

Furthermore, a previous report suggests that T-cell prolif-eration in autoimmunity is stimulated by leptin.16 Our current findings are in accordance with a previous report by Yoshikawa et al (1985).17 FCA-induced arthritic rats failed to gain weight; however, this was significantly attenuated in arthritic rats treated with PCP, resulting in increased body weight in a dose-dependent manner.

Hyperalgesia is associated with the inflammatory response due to bone deformation and joint disability in arthritis.18 To determine the extent of hyperalgesia, DFP, motility and stair climbing ability were tested in arthritic rats. Treatment with PCP produced improvement in all three of these measures in FCA-induced arthritic rats. The anti-nociceptive effect of PCP could be attributed to the inhibition of prostaglandins and cytokines.13

The thymus and spleen are vital organs involved in im-mune responses. Rats treated with PCP exhibited a marked reduction in thymus and spleen indices. These results indicate that PCP may affect the immune function of arthritic rats.

Elevated leucocyte levels play a major role in the body’s defence mechanism. The inflammatory response to FCA-induced arthritis causes increased numbers of granulocytes and macrophages, proportional to the elevated leucocyte levels.16PCP administration plays a role in the inhibition of inflammatory mediators that may decrease total leucocyte levels in arthritic rats. However, there were no significant differences observed in the haematological profile of PCP-treated rats.

Table 1 Effect of PCP on immune organs in arthritic rats. Treatment Dose


Thymus index Spleen index Control e 0.44 (0.010) 2.71 (0.080) FCA Control e 0.62 (0.012)# 3.60 (0.090)# Indomethacin 1 0.48 (0.013)*** 2.78 (0.047)*** PCP 100 0.56 (0.014)* 3.60 (0.067)** 200 0.52 (0.014)*** 2.98 (0.084)*** 400 0.45 (0.018)*** 2.93 (0.065)***

Values are presented as the mean (SEM), nZ 6.

*P < .05, **P < .01 and ***P < .001 compared with the FCA control group.

#P< .05 compared with vehicle control.

Table 2 Effect of PCP on total leucocyte count, differential leucocyte count and platelet count of arthritic rats. Treatment Dose


Haematological parameters (SEM) Total count (103/cmm) Neutrophils (%) Eosinophils (%) Lymphocytes (%) Monocytes (%) Platelets (105/mL) Control e 9.40 (0.560) 24.33 (2.171) 2.33 (0.333) 72.67 (1.978) 0.67 (0.211) 8.98 (0.499) FCA Control e 11.97 (0.547) 31.67 (2.275) 3.67 (0.703) 64.33 (1.928) 0.33 (0.211) 9.38 (0.856) Indomethacin 1 8.70 (0.546)** 25.33 (1.706) 3.17 (0.477) 71.00 (1.506) 0.50 (0.342) 8.62 (0.521) PCP 100 10.58 (0.498) 27.33 (2.390) 2.33 (0.615) 70.00 (2.595) 0.33 (0.211) 8.97 (0.518) 200 10.47 (0.544) 26.67 (2.186) 2.17 (0.477) 70.50 (1.928) 0.67 (0.333) 9.75 (0.423) 400 9.68 (0.693)* 26.00 (2.324) 1.83 (0.307) 72.00 (2.113) 0.17 (0.167) 7.22 (0.633)

Values are presented as the mean (SEM), nZ 6.


Oxidative stress is increased in the liver of arthritic rats. Downregulation of the endogenous antioxidant defence sys-tem leads to destruction of the joint structure due to gener-ation of free radicals in FCA-induced arthritis.19 Administration of PCP leads to restoration of free radical

scavenging ability. The upregulation of SOD, CAT and GSH in the liver is an indication of the restoration of the antioxidant defence system.11MDA content is an index of exaggerated peroxidation process. In the present study, enhanced oxida-tive stress was clearly evidenced by marked increases in MDA

Figure 5 Effect of PCP on indices of oxidative stress in liver of arthritic rats. Values are presented as the mean (SEM), nZ 6. *P< .05, **P < .01 and ***P < .001 compared with the FCA control group.#P< .05 compared with vehicle control.


and NO activity in arthritic rats. Increased production of NO is mediated by enhanced iNOS expression in the inflamed joints.20Obstruction in growth and differentiation of osteo-blasts occurs because of high concentrations of NO, which may be responsible for the inhibitory effect on bone forma-tion in the arthritic condiforma-tion.11 Treatment with PCP sup-pressed the rise of MDA and NO in liver, indicating a protective effect, and restored altered indicators towards normal levels. Thus, PCP may exert a stabilising effect by protecting bio-logical membranes from oxidative damage.

Increased aminotransferase enzymes (AST, ALT) and ALP activities were also observed in arthritic rats. An increase in these cellular enzyme levels is an index of organ and bone impairment. AST and ALT affect the formation of mediators in inflammatory processes, while increased ALP levels affect bone loss in the form of bone erosion, which in turn results in damage to organs and bone.21,22 Hence, the decreased levels of these indices following PCP treatment indicate an effective anti-arthritic activity by protecting against organ and bone erosion.

Elevated levels of inflammatory cytokines (IL-6 and TNF-a) are observed in the early phase of the arthritic condi-tion.23 It is believed that inflammatory mediators play an important role in pathogenesis of arthritis by inducing infiltration of immune cells. TNF-a stimulates the produc-tion of other inflammatory cytokines such as 1 and IL-6.11,24 Similarly, IL-6 contributes to symptoms associated with arthritis, including T-cell and B-cell proliferation and differentiation, antibody production and osteoclast acti-vation.25 In agreement with this, our results demonstrate that PCP exerts an anti-arthritic effect by suppressing the production of TNF-a and IL-6 induced by administration of FCA in rats.

Histopathological examination of tibiotarsal thickness of adjuvant arthritis rats showed that the higher doses of 200 and 400 mg/kg PCP are effective in reducing the number of infiltrating inflammatory cells, with improvement in bone resorption. Inhibition of TNF-a suppresses joint swelling and synovial inflammation while IL-6 inhibition leads to protection of cartilage and bone.26This study demonstrates Figure 6 Effect of PCP on levels of tissue marker enzymes Aspartate transaminase (AST), Alanine transaminase (ALT), Alkaline phosphatase (ALP) in arthritic rats serum. Values are presented as the mean (SEM), nZ 6. *P < .05, **P < .01 and ***P < .001 compared with the FCA control group.#P< .05 compared with vehicle control.

Figure 7 Effect of PCP on the inflammatory cytokines, TNF-a and IL-6 in plasma samples. Values are presented as the mean (SEM), nZ 6. yP < .05 compared with the FCA control group.#P< .05 compared with vehicle control.


the PCP-mediated inhibition of inflammatory cell infiltra-tion and preveninfiltra-tion of bone resorpinfiltra-tion.

In summary, the above findings reveal that the PCP fraction of C. paniculatus possesses potential anti-arthritic properties. The underlying mechanism for the beneficial effects of PCP could be mediated through several factors such as regulation of anti-inflammatory cytokines, allevi-ating oxidative stress, increased cellular enzymes and organ and bone-protective activities. Further research needs to be carried out to elucidate the more detailed molecular mechanisms involved in these novel properties of C. paniculatus.

Conflict of interest

The authors declare that there are no conflicts of interest.


The authors are grateful for the financial support from the University Grant Commission (UGC), India (No. F 5-63/ 2007). The authors also wish to thank Dr Dnyaneshwar Nagmoti, Mr Nitin Gawali and Ms Sarayu Pai who contrib-uted to this study.


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C: carƟlage; S: synovium; B: bone; SM: synovial membrane; I: infiltraƟon; JS: joint space; CD: carƟlage damage; BD: bone damage; H: hyperplasia

V JS SM S FCA H C I Indo SM PCP 400 SM C B S PCP 200 C B S PCP 100 H

Figure 8 Effect of PCP on histopathological changes of tibiotarsal joints in arthritic rats. All images are from one of six in each group. V: vehicle control; FCA: adjuvant-induced arthritic control; Indo: indomethacin (1 mg/kg); PCP 100, 200 and 400 indicate different doses of PCP at 100, 200 and 400 mg/kg, respectively.


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