Korean J Physiol Pharmacol.  2020 Nov;24(6):453-462. 10.4196/kjpp.2020.24.6.453.

Anti-arthritic activity of D-carvone against complete Freund’s adjuvant-induced arthritis in rats through modulation of inflammatory cytokines

• Chen G
• Song Y
• Ma F
• Ma Y

• KMID: 2507728
• DOI: http://doi.org/10.4196/kjpp.2020.24.6.453

Abstract

Chronic joint pain due to loss of cartilage function, degradation of subchondral bone, and related conditions are common plights of an arthritis patient. Antioxidant compounds could solve the problems in arthritic condition. The objective of this study was to evaluate the anti-arthritic activity of D-carvone against complete Freund’s adjuvant (CFA)-induced arthritis in rats. D-carvone was orally administered for 25 days at the doses of 30 and 60 mg/kg against CFA-induced arthritic rats. Changes in body weight, paw swelling, organ index, hematological parameters, oxidative stress markers, inflammatory cytokines, and histopathology were recorded. Oral treatment of D-carvone significantly improved the body weight, reduced the paw swelling, edema formation, and organ index in arthritic rats. The levels of white blood cells were reduced, red blood cells and hemoglobin levels were improved in D-carvone treated arthritic rats. Lipid peroxidation levels were lowered whereas enzymatic and non-enzymatic antioxidants were significantly elevated by D-carvone administration against arthritic rats. D-carvone significantly modulated inflammatory cytokine levels and improved the ankle joint pathology against CFA-induced arthritic inflammation. In conclusion, D-carvone proved significant anti-arthritic activity against CFA-induced arthritis in rats.

Keyword

Arthritis; D-carvone; Inflammatory cytokines; Oxidative stress

Figure

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  Fig. 1 D-carvone reduces the hind paw volume of CFA-induced arthritic rats. Values expressed as mean ± standard error of mean of (n = 8) rats. Group I = Normal control; Group II = CFA-induced arthritis model; Group III = D-carvone 30 mg/kg b.w. + CFA; Group IV = D-carvone 60 mg/kg b.w. + CFA; Group V = Indomethacin positive control + CFA. CFA, complete Freund’s adjuvant; b.w., body weight. Superscript ‘a’ indicates significant difference (p < 0.05) from Group I, superscript ‘b’ indicates significant difference (p < 0.05) from Group II, superscript ‘c’ indicates significant difference (p < 0.05) from Group III, by one-way ANOVA followed by Tukey’s multiple comparisons test.

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  Fig. 2 D-carvone alters the arthritic score of CFA-induced inflammation in rats. Values expressed as mean ± standard error of mean of (n = 8) rats. The arthritic score was ranged from 0 (normal paw with no swelling or erythema)–4 (severe deformity of the paw with swelling and erythema) for a maximum score of 8 for both hind paws. Group I = Normal control; Group II = CFA-induced arthritis model; Group III = D-carvone 30 mg/kg b.w. + CFA; Group IV = D-carvone 60 mg/kg b.w. + CFA; Group V = Indomethacin positive control + CFA. CFA, complete Freund’s adjuvant; b.w., body weight. Superscript ‘a’ indicates significant difference (p < 0.05) from Group I, superscript ‘b’ indicates significant difference (p < 0.05) from Group II, superscript ‘c’ indicates significant difference (p < 0.05) from Group III, by one-way ANOVA followed by Tukey’s multiple comparisons test.

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  Fig. 3 Effects of D-carvone on serum liver markers in CFA-induced arthritic rats. Values expressed as mean ± standard error of mean (n = 8) of hematological changes due to D-carvone administration in CFA-induced rats. Group I = Normal control; Group II = CFA-induced arthritis model; Group III = D-carvone 30 mg/kg b.w. + CFA; Group IV = D-carvone 60 mg/kg b.w. + CFA; Group V = Indomethacin positive control + CFA. CFA, complete Freund’s adjuvant; b.w., body weight; ALP, alkaline phosphatase; AST, aspartate aminotransferase; ALT, alanine aminotransferase. Superscript ‘a’ indicates significant difference (p < 0.05) from Group I, superscript ‘b’ indicates significant difference (p < 0.05) from Group II, superscript ‘c’ indicates significant difference (p < 0.05) from Group III, by one-way ANOVA followed by Tukey’s multiple comparisons test.

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  Fig. 4 Effects of D-carvone on oxidative stress in CFA-induced arthritic rats. Values expressed as mean ± standard error of mean (n = 8) of lipid peroxidation, enzymatic and non-enzymatic antioxidants. Group I = Normal control; Group II = CFA-induced arthritis model; Group III = D-carvone 30 mg/kg b.w. + CFA; Group IV = D-carvone 60 mg/kg b.w. + CFA; Group V = Indomethacin positive control + CFA. CFA, complete Freund’s adjuvant; b.w., body weight; MDA, malondialdehyde; GSH, reduced glutathione; SOD, superoxide dismutase; CAT, catalase. Superscript ‘a’ indicates significant difference (p < 0.05) from Group I, superscript ‘b’ indicates significant difference (p < 0.05) from Group II, superscript ‘c’ indicates significant difference (p < 0.05) from Group III, by one-way ANOVA followed by Tukey’s multiple comparisons test.

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  Fig. 5 Effects D-carvone on inflammatory cytokines in CFA-induced arthritic rats. Values expressed as mean ± standard error of mean (n = 8) of serum inflammatory cytokines. Group I = Normal control; Group II = CFA-induced arthritis model; Group III = D-carvone 30 mg/kg b.w. + CFA; Group IV = D-carvone 60 mg/kg b.w. + CFA; Group V = Indomethacin positive control + CFA. CFA, complete Freund’s adjuvant; b.w., body weight; IL, interleukin; TNF, tumor necrosis factor. Superscript ‘a’ indicates significant difference (p < 0.05) from Group I, superscript ‘b’ indicates significant difference (p < 0.05) from Group II, superscript ‘c’ indicates significant difference (p < 0.05) from Group III, by one-way ANOVA followed by Tukey’s multiple comparisons test.

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  Fig. 6 Modulating effect of D-carvone on the hind paw histopathology of CFA-induced rats. (A) Hind paw sections of arthritis model group II shows severe signs of articular cartilage damage, infiltration of inflammatory cells, synovial hyperplasia, and bone marrow degeneration. Administration of D-carvone in rats of group III and IV ameliorated the CFA-induced arthritic inflammation and changes. Positive control indomethacin group V also exhibited positive changes against CFA-induced arthritic damage. Arrow indicates articular cartilage erosion. Scale bar = 500 µm; H&E staining at 100× magnification. (B) Total Mankin’s score for articular injuries and cellular irregularities. Group I = Normal control; Group II = CFA-induced arthritis model; Group III = D-carvone 30 mg/kg b.w. + CFA; Group IV = D-carvone 60 mg/kg b.w. + CFA; Group V = Indomethacin positive control + CFA. CFA, complete Freund’s adjuvant; b.w., body weight. Superscript ‘a’ indicates significant difference (p < 0.05) from Group I, superscript ‘b’ indicates significant difference (p < 0.05) from Group II, superscript ‘c’ indicates significant difference (p < 0.05) from Group III, by one-way ANOVA followed by Tukey’s multiple comparisons test.

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