Anti-arthritic activity of Ricinus communis L. and Withania somnifera L. extracts in adjuvant-induced arthritic rats via modulating inflammatory mediators and subsiding oxidative stress

Document Type : Original Article


Institute of Physiology and Pharmacology, University of Agriculture, Faisalabad-38040, Faisalabad, Punjab, Pakistan


Objective(s): This study aimed to evaluate the anti-arthritic activity of Ricinus communis leaves’ and Withania somnifera roots’ hydroalcoholic extracts in Complete Freund’s adjuvant-induced arthritis in Wistar rats.
Materials and Methods: HPLC and FT-IR analysis detected pharmacologically important phytocompounds in both plant extracts. Oral treatments including methotrexate (MTX; 3 mg/kg twice a week) and extracts at 250 and 500 mg/kg/day were initiated after arthritis induction. Changes in paw swelling, arthritic score, body weight, organ indices (thymus and spleen), hematological and biochemical parameters, and pro-/anti-inflammatory cytokine expression using qRT-PCR were assessed. Oxidative stress markers in hepatic tissue were determined. Histopathological and radiological examinations were also performed.  
Results: RCE (R. communis extract) and WSE (W. somnifera extract) demonstrated a reduction in paw swelling, arthritic score, and restoration of body weight and organ indices. Hematological parameters, serum inflammatory markers such as CRP and RF, and liver function markers of arthritic rats were significantly (p <0.01) ameliorated with RCE and WSE treatment. Both plants persuasively down-regulated IL-1β, IL-6, IL-17a, TNF-α, and RANKL and up-regulated IL-4, INF-γ, and OPG relative expression as well as alleviating hepatic oxidative stress parameters. Histopathological and radiological findings revealed a marked reduction in tissue inflammation and bone erosion in extracts treated groups.
Conclusion: The study findings suggest that R. communis leaves and W. somnifera roots have markedly subsided inflammation and improved health through modulating pro-/anti-inflammatory cytokine expression and reducing oxidative stress.


1.    Giannini D, Antonucci M, Petrelli F, Bilia S, Alunno A, Puxeddu I. One year in review 2020: pathogenesis of rheumatoid arthritis. Clin Exp Rheumatol 2020; 38:387–397.
2.    Naqvi AA, Hassali MA, Aftab MT. Epidemiology of rheumatoid arthritis, clinical aspects and socio-economic determinants in Pakistani patients: A systematic review and meta-analysis. J Pak Med Assoc 2019; 69:389–398.
3.    Cho SK, Kim D, Won S, Lee J, Choi CB, Choe JY, et al. Factors associated with time to diagnosis from symptom onset in patients with early rheumatoid arthritis. Korean J Intern Med 2019; 34:910-916.
4.    Scherer HU, Häupl T, Burmester GR. The etiology of rheumatoid arthritis. J Autoimmun 2020; 110:e102400.
5.    Phull AR, Nasir B, ul Haq I, Kim SJ. Oxidative stress, consequences and ROS mediated cellular signaling in rheumatoid arthritis. Chem Biol Interact 2018; 281:121–136.
6.    Fonseca LJ, Nunes-Souza V, Goulart MO, Rabelo LA. Oxidative stress in rheumatoid arthritis: What the future might hold regarding novel biomarkers and add-on therapies. Oxid Med Cell Longev 2019; e31934269.    
7.    Chen Z, Bozec A, Ramming A, Schett G. Anti-inflammatory and immune-regulatory cytokines in rheumatoid arthritis. Nat Rev Rheumatol 2019; 15:9–17.  
8.    Croia C, Bursi R, Sutera D, Petrelli F, Alunno A, Puxeddu I. One year in review 2019: pathogenesis of rheumatoid arthritis. Clin Exp Rheumatol 2019; 37:347–357.
9.    Alivernini S, Tolusso B, Fedele AL, Di Mario C, Ferraccioli G, Gremese E. The B side of rheumatoid arthritis pathogenesis. Pharmacol Res 2019; 149:104465.
10.    Miossec P. Local and systemic effects of IL-17 in joint inflammation: A historic perspective from discovery to targeting. Cell Mol Immunol 2021; 18:860–865.
11.    Dong C, Ting F, Juan J, Zhenyu L, Zhifeng G. The role of interleukin-4 in rheumatic diseases. Clin Exp Pharmacol Physiol 2018; 45:747–754.
12.    Lee SH, Ji YK, Se-Young K, Kyoung AJ, Mi-La C. Interferon-gamma regulates inflammatory cell death by targeting necroptosis in experimental autoimmune arthritis. Sci Rep 2017; 7:e10133.
13.    Favalli EG. Understanding the role of interleukin-6 (il-6) in the joint and beyond: A comprehensive review of il-6 inhibition for the management of rheumatoid arthritis. Rheumat Ther 2020; 7:473–516.   
14.    Metzger CE, Narayanan SA. The role of osteocytes in inflammatory bone loss. Front Endocrinol 2019; 10:285.  
15.    Köhler BM, Günther J, Kaudewitz D, Lorenz HM. Current therapeutic options in the treatment of rheumatoid arthritis. J Clin Med 2019; 8:938 PMC6678427.
16.    Costello R, David T, Jani M. Impact of adverse events associated with medications in the treatment and prevention of rheumatoid arthritis. Clin Ther 2019; 41:1376–1396.   
17.    Li TP, Zhang AH, Miao JH, Sun H, Yan GL, Wu FF, et al. Applications and potential mechanisms of herbal medicines for rheumatoid arthritis treatment: A systematic review. RSC Adv 2019; 9:26381–26392.   
18.    Abdul WM, Hajrah NH, Sabir JS, Al-Garni SM, Sabir MJ, Kabli SA, et al. Therapeutic role of Ricinus communis L. and its bioactive compounds in disease prevention and treatment. Asian Pac J Trop Med 2018; 11:177-185.
19.    Babu PR, Bhuvaneswar C, Sandeep G, Ramaiah CV, Rajendra W. Hepatoprotective role of Ricinus communis leaf extract against d-galactosamine induced acute hepatitis in albino rats. Biomed Pharmacother 2017; 88:658–666.  
20.    Farooq U, Khan A, Naz S, Rauf A, Khan H, Khan A, et al. Sedative and antinociceptive activities of two new sesquiterpenes isolated from Ricinus communis. Chin J Nat Med 2018; 16:225–230.   
21.    Javanshir A, Karimi E, Maragheh AD, Tabrizi MH. The anti-oxidant and anticancer potential of Ricinus communis L. essential oil nanoemulsions. J Food Meas Charact 2020; 1–10.   
22.    Kalra R, Kaushik N. Withania somnifera (Linn.) Dunal: A review of chemical and pharmacological diversity. Phytochem Rev 2017; 16:953–987.  
23.    Dhanani T, Shah S, Gajbhiye NA, Kumar S. Effect of extraction methods on yield, phytochemical constituents and anti-oxidant activity of Withania somnifera. Arab J Chem 2017; 10:S1193–S1199.   
24.    Saleem S, Muhammad G, Hussain MA, Altaf M, Bukhari SN. Withania somnifera L.: Insights into the phytochemical profile, therapeutic potential, clinical trials, and future prospective. Iran J Basic Med Sci 2020; 23:1501–1526.  
25.    Seal T. Quantitative HPLC analysis of phenolic acids, flavonoids and ascorbic acid in four different solvent extracts of two wild edible leaves, Sonchus arvensis and Oenanthe linearis of North-Eastern region in India. J Appl Pharma Sci 2016; 6:157–166.  
26.    Zulfqar F, Akhtar MF, Saleem A, Akhtar B, Sharif A, Saleem U. Chemical characterization, anti-oxidant evaluation, and antidiabetic potential of Pinus gerardiana (Pine nuts) extracts. J Food Biochem 2020; 44:e13199.  
27.    Kareru P, Keriko J, Gachanja A, Kenji G. Direct detection of triterpenoid saponins in medicinal plants. Afr J Tradit Complement Altern Med 2008; 5:56–60.
28.    Gad-Elkareem MA, Abdelgadir EH, Badawy OM, Kadri A. Potential antidiabetic effect of ethanolic and aqueous-ethanolic extracts of Ricinus communis leaves on streptozotocin-induced diabetes in rats. PeerJ 2019; 7:e6441.   
29.    Patel SB, Rao NJ, Hingorani LL. Safety assessment of Withania somnifera extract standardized for Withaferin A: Acute and sub-acute toxicity study. J Ayurveda Integr Med 2016; 7:30–37.  
30.    Zhang X, Dong Y, Dong H, Zhang W, Li F. Investigation of the effect of phlomisoside F on complete Freund’s adjuvant-induced arthritis. Exp Ther Med 2017; 13:710–716.   
31.    Luo S, Li H, Liu J, Xie X, Wan Z, Wang Y, et al. Andrographolide ameliorates oxidative stress, inflammation and histological outcome in complete Freund’s adjuvant-induced arthritis. Chem Biol Interact 2020; 319 e108984.   
32.    Zhang RX, Fan AY, Zhou AN, Moudgil KD, Ma ZZ, Lee DYW, et al. Extract of the Chinese herbal formula Huo Luo Xiao Ling Dan inhibited adjuvant arthritis in rats. J Ethnopharmacol 2009; 121:366–371.  
33.    Zhang Q, Yu Y, Li J, Guan Y, Huang J, Wang Z, et al. Anti-arthritic activities of ethanol extracts of Circaea mollis Siebold & Zucc. (whole plant) in rodents. J Ethnopharmacol 2018; 225:359–366.  
34.    Coura CO, Souza RB, Rodrigues JAG, Vanderlei EDSO, de-Araújo IWF, Ribeiro NA, et al. Mechanisms involved in the anti-inflammatory action of a polysulfated fraction from Gracilaria cornea in rats. PLoS One 2015; 10:e0119319.  
35.    Adeyemi WJ, Olayaki LA. Diclofenac–induced hepatotoxicity: low dose of omega-3 fatty acids have more protective effects. Toxicol Rep 2018; 5:90–95.       
36.    Sharif A, Ashraf M, Javeed A, Anjum AA, Akhtar MF, Akhtar B. Oxidative stress responses in Wistar rats on subacute exposure to pharmaceutical wastewater. Environ Sci Pollut Res Int 2016; 23:24158–24165.  
37.    Muller SD, Florentino D, Ortmann CF, Martins FA, Danielski LG, Michels M, et al. Anti-inflammatory and anti-oxidant activities of aqueous extract of Cecropia glaziovii leaves. J Ethnopharmacol 2016; 185:255–262.      
38.    Ananthia S, Gayathria V, Malarvizhib R, Bhardwajb M, Vasanthi HR. Anti-arthritic potential of marine macroalgae Turbinaria ornata in Complete Freund’s Adjuvant induced rats. Exp Toxicol Pathol 2017; 69:672–680.       
39.    Oliveira RN, Mancini MC, Oliveira FC, Passos TM, Quilty B, Thiré RM, et al. FTIR analysis and quantification of phenols and flavonoids of five commercially available plants extracts used in wound healing. Matéria (Rio J) 2016; 21:767–779.   
40.    Foyet HS, Tsala DE, Bodo JZE, Carine AN, Heroyne LT, Oben EK. Anti-inflammatory and anti-arthritic activity of a methanol extract from Vitellaria paradoxa stem bark. Pharmacog Res 2015; 7:367–377.       
41.    Taksande BG, Gawande DY, Chopde CT, Umekar MJ, Kotagale NR. Agmatine ameliorates adjuvant induced arthritis and inflammatory cachexia in rats. Biomed Pharmacother 2017; 86:271–278.  
42.    Issekutz AC, Sapru K. Modulation of adjuvant arthritis in the rat by 2-methoxyestradiol: an effect independent of an anti-angiogenic action. Int Immunopharmacol 2008; 8:708–716.  
43.    Wang, S, Wang Y, Liu X, Guan L, Yu L, Zhang X. Anti-inflammatory and anti-arthritic effects of taraxasterol on adjuvant-induced arthritis in rats. J Ethnopharmacol 2016; 187:42–48.  
44.    Thite AT, Patil RR, Naik SR. Anti-arthritic activity profile of methanolic extract of Ficus bengalensis: comparison with some clinically effective drugs. Biomed Aging Pathol 2014; 4:207–217.  
45.    Moon SJ, Ahn IE, Kwok SK, Park KS, Min JK, Park SH, et al. Periarticular osteoporosis is a prominent feature in early rheumatoid arthritis: estimation using shaft to periarticular bone mineral density ratio. J Korean Med Sci 2013; 28:287–294.  
46.    Kalaiselvan S, Rasool MK. Triphala herbal extract suppresses inflammatory responses in LPS-stimulated RAW 264.7 macrophages and adjuvant-induced arthritic rats via inhibition of NF-κB pathway. J Immunotoxicol 2016; 13:509–525.
47.    Altindag O, Karakoc M, Kocyigit A, Celik H, Soran N. Increased DNA damage and oxidative stress in patients with rheumatoid arthritis. Clin Biochem 2007; 40:167–171.
48.    Voon FL, Sulaiman MR, Akhtar MN, Idris MF, Akira A, Perimal EK, et al. Cardamonin (2′, 4′-dihydroxy-6′-methoxychalcone) isolated from Boesenbergia rotunda (L.) Mansf. inhibits CFA-induced rheumatoid arthritis in rats. Eur J Pharmacol 2017; 794:127–134.
49.    Srirangan S, Choy EH. The role of interleukin 6 in the pathophysiology of rheumatoid arthritis. Ther Adv Musculoskelet Dis 2010; 2:247–256.
50.    Yen FL, Wu TH, Lin LT, Lin CC. Hepatoprotective and anti-oxidant effects of Cuscuta chinensis against acetaminophen-induced hepatotoxicity in rats. J Ethnopharmacol 2007; 111:123–128.
51.    Gerards AH, de Lathouder S, De Groot ER, Dijkmans BAC, Aarden LA. Inhibition of cytokine production by methotrexate. Studies in healthy volunteers and patients with rheumatoid arthritis. Rheumatology 2003; 42:1189–1196.