Abstract
Adjuvant arthritis is a chronic, autoimmune and inflammatory disorder of the joints. The occurrence of disorder causes a severe damage to the connective tissue eventually leading to progressive physical disability and eventual death. The recent years of evidence suggests the anti-inflammatory properties of stevioside, a diterpene glycoside. However, the effect of stevioside against adjuvant arthritis, a chronic inflammatory disorder is not known. Hence, the present study was designed to study the effect of stevioside against Freund's complete adjuvant induced arthritis model in rats. The acute anti-inflammatory effect of stevioside also studied by employing carrageenan-induced paw oedema model in rats. The biochemical markers, haematological parameters, lipid peroxidation, myeloperoxidase activity, lipoxygenase activity, the levels of PGE2 and pro-inflammatory (TNF-α, IL-6 & IL-1β) and anti-inflammatory cytokine (IL-10) were analysed. The protein expression of NF-κB (p65) COX-2 and iNOS in paw tissues were estimated by western blotting. Stevioside treatment significantly ameliorates the adjuvant induced arthritic scoring, histological alterations, paw volume, elevation of biochemical (AST, ALT, ALP and glucose levels) and haematological (haemoglobin, differential and platelet count) parameters and restored the endogenous anti-oxidant (SOD, CAT, GSH and GST) activities. Treatment with stevioside also significantly prevented the adjuvant induced elevation of pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β), pro-inflammatory protein expressions (iNOS, COX-2, NF-κB (p65) and pIκB/IκB ratio), prevented the increase in myeloperoxidase activity and significantly restored the anti-inflammatory (IL-10) cytokine level in paw tissues. Collectively, our findings suggest that stevioside may serve as anti-inflammatory agent and could serve as a potential adjunct therapeutic option in treating adjuvant arthritis.
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Abbreviations
- ANOVA:
-
Analysis of variance
- BCA:
-
Bicinchoninic acid
- FCA:
-
Freund’s complete adjuvant
- RIPA:
-
Radioimmunoprecipitation assay
- PVDF:
-
Polyvinylidine difluoride
- BSA:
-
Bovine serum albumin
- TBA:
-
2-Thiobarbituric acid
- H&E:
-
Hematoxylin and eosin staining
- DTNB:
-
5,5′-Dithio-bis-(2-nitrobenzoic acid)
- CDNB:
-
1-Chloro-2,4-dinitrobenzene
- COX-2:
-
Cyclooxygenase-2
- TNF-α:
-
Tumour necrosis factor-α
- IL-6:
-
Interleukin-6
- IL-1β:
-
Interleukin-1β
- IL-10:
-
Interleukin-10
- NF-κB:
-
Nuclear factor kappa B
- IκB:
-
Inhibitor of kappa B
- iNOS:
-
Inducible nitric oxide synthase
- MDA:
-
Malondialdehyde
- TBARS:
-
Thiobarbituric acid reactive substances
- ROS:
-
Reactive oxygen species
- SEM:
-
Standard error of mean
- CAT:
-
Catalase
- SOD:
-
Superoxide dismutase
- HRP:
-
Horseradish peroxidise
- ELISA:
-
Enzyme-linked immunosorbent assay
- p.o:
-
Per oral
- PBS:
-
Phosphate buffered saline
- HTAB:
-
Hexadecyltrimethylammonium bromide
- AST:
-
Aspartate aminotransferase
- ALT:
-
Alanine aminotransferase
- ALP:
-
Alkaline phosphatase
- GST:
-
Glutathione S-transferase
- PGE2:
-
Prostaglandin E2
- SDS-PAGE:
-
Sodium dodecyl sulfate–polyacrylamide gel electrophoresis
- EDTA:
-
Ethylenediaminetetraacetic acid
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Acknowledgements
SA acknowledges the Department of Science and Technology (DST), New Delhi, India, for financial assistance in the form of Inspire Fellowship (IF160504). NN and SR thank the Indian Council of Medical Research (ICMR), New Delhi, India and the Council of Scientific and Industrial Research (CSIR) (Grant No. 31/014(2683)/2016-EMR-I), New Delhi, India, respectively, for the award of Senior Research Fellowships (SRF). We thank Director CSIR-IICT for the support (IICT/Pubs./2019/413).
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Alavala, S., Nalban, N., Sangaraju, R. et al. Anti-inflammatory effect of stevioside abates Freund’s complete adjuvant (FCA)-induced adjuvant arthritis in rats. Inflammopharmacol 28, 1579–1597 (2020). https://doi.org/10.1007/s10787-020-00736-0
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DOI: https://doi.org/10.1007/s10787-020-00736-0