Abstract
Background
Rheumatoid arthritis (RA) is a chronic inflammatory disorder causing cartilage and joint degeneration. In spite of the availability of several robust drugs like biologics, most of the patients are unresponsive, and reports of severe adverse effects following long-term use are also there. Subsequently the use of natural plant-based products in RA therapy is broadening over the years. Tinospora cordifolia is a widely used medicinal plant in Ayurveda against various inflammatory disorders including RA. However, there is very limited knowledge regarding the actual molecular events responsible for its therapeutic effect, and this has limited its acceptance among the professionals.
Purpose
To explore the anti-inflammatory and anti-arthritic effect of hydro-alcoholic extract from Tinospora cordifolia.
Methods
The rich polyphenol nature of the extract was elucidated using HPLC. LPS-stimulated murine macrophage cell line RAW 264.7 was used for in vitro studies, and collagen-induced arthritis (CIA) model was used for in vivo studies.
Results
The polyphenols in TCE were identified using HPLC. TCE effectively downregulated the level of pro-inflammatory mediators (IL-6, TNF-α, PGE2, and NO) in LPS-stimulated RAW 264.7 cells. Subsequently the upregulated expression of COX-2 and iNOS following LPS stimulation were also downregulated by TCE. Furthermore, TCE targeted the upstream kinases of the JAK/STAT pathway, a crucial inflammatory pathway. The expression of VEGF, a key angiogenic factor as well as an inflammatory mediator was also decreased following pre-treatment with TCE. The anti-arthritic effect of TCE (150 mg/kg) was evaluated in the CIA model as well. From the results of histopathology, oral administration of TCE was found to be effective in reducing the clinical symptoms of arthritis including paw edema, erythema, and hyperplasia. In vivo results validated the in vitro results and there was a significant reduction in serum level of pro-inflammatory cytokines and mediators (IL-6, TNF-α, IL-17, NO, and PGE2). The phosphorylation of STAT3 and the expression of VEGF were also downregulated following TCE treatment.
Conclusion
Our study provided a detailed insight into the molecular events associated with anti-inflammatory and anti-arthritic effect of Tinospora cordifolia.
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Data availability
The datasets generated or analyzed for the present study are available from the corresponding author upon reasonable request.
Abbreviations
- TCE:
-
Tinospora cordifolia Extract
- ELISA:
-
Enzyme-linked immunosorbent assay
- LPS:
-
Lipopolysaccharides
- ROS:
-
Reactive oxygen species
- NSAIDs:
-
Non-steroidal anti-inflammatory drugs
- DMARDs:
-
Disease-modifying anti-rheumatic drugs
- bDMARDs:
-
Biological disease-modifying anti-rheumatic drugs
- CIA:
-
Collagen-induced arthritis animal model
- CFA:
-
Complete Freund’s adjuvant
- JAK/STAT:
-
Janus kinase/signal transducers and activators of transcription
- VEGF:
-
Vascular endothelial growth factor
- RA:
-
Rheumatoid arthritis
- DMEM:
-
Dulbecco's modified Eagle's medium
- FBS:
-
Fetal bovine serum
- BSA:
-
Bovine serum albumin
- HPLC:
-
High-performance liquid chromatography
- SDS:
-
Sodium dodecyl sulfate
- HBSS:
-
Hanks balanced saline solution
- PBS:
-
Phosphate-buffered saline
- MTT:
-
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide
- DCFH-DA:
-
2, 7-Dichlorodihydrofluorescein diacetate
- RIPA:
-
Radio-immunoprecipitation assay
- ATCC:
-
American Type Culture Collection
- Dex:
-
Dexamethasone
- DMSO:
-
Dimethyl sulfoxide
- NO:
-
Nitric oxide
- DAPI:
-
4′, 6-Diamidino-2-phenylindole
- PVDF:
-
Polyvinylidene difluoride
- ECL:
-
Enhanced chemiluminescence
- DAS-28:
-
Disease activity score 28
- CPCSEA:
-
Committee for the purpose of control and supervision of experiments on animals
- IAEC:
-
Institutional animal ethics committee
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Acknowledgements
Dr. Genu George would like to acknowledge ICMR, India for the financial support as fellowship (2020-3899, No.3/1/2(1)/CD/2021-NCD-II). We acknowledge the Director of CSIR-NIIST, Thiruvananthapuram for granting all the necessary facilities. We also thank the Director, Jubilee Mission Medical College and Research Institute, Thrissur for providing the necessary facilities for conducting in vivo experiments.
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Dr. K.G.R conceived and designed the idea. Dr. GG conducted the entire in vitro and in vivo study. Manuscript drafting and editing was done by Dr. GG, and Dr. K.G.R. Dr. G.L.S, and Dr. SM, assisted in the in vivo study. Dr. K.R, and Dr. S. R contributed in the design and conduct of in vivo experiments. BA and Dr. P.N have done the polyphenol characterization of the Tinospora cordifolia extract.
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All the in vivo experiments were conducted at small animal research facility, Jubilee Mission Medical College and Research Institute, Thrissur in strict compliance to the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India (RP-03/2017(i)/IAEC/JMMC&RI).
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George, G., Shyni, G.L., Mohan, S. et al. In vitro and in vivo anti-inflammatory and anti-arthritic effect of Tinospora cordifolia via modulation of JAK/STAT pathway. Inflammopharmacol 31, 1009–1025 (2023). https://doi.org/10.1007/s10787-023-01155-7
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DOI: https://doi.org/10.1007/s10787-023-01155-7