Biochemical Evaluation of Withania Somnifera Root Powder on Adjuvant-Induced Arthritis in Rats

The present investigation was carried out to evaluate the biochemical effect of Withania somnifera Linn. Solanaceae, commonly known as ashwagandha on adjuvant induced arthritic rats. Results were compared to Indomethacin, a non steroidal anti-inflammatory drug. Arthritis was induced by an intra dermal injection of Complete Freund’s Adjuvant (0.1 ml) into the right hind paw of Wistar albino rats. Withania somnifera root powder (1000 mg/kg/day) and Indomethacin (3 mg/kg/day) were orally administered for 8 days (from 11th to 18th day) after adjuvant injection. After the experimental period, all the animals were sacrificed and serum, liver and spleen samples were collected for further biochemical analysis. A significant increase in the activities of gluconeogenic enzymes, tissue marker enzymes, blood glucose level, WBC, platelet count, erythrocyte sedimentation rate, and acute phase proteins (hyaluronic acid, fibrinogen and ceruloplasmin) was observed in adjuvant-induced arthritic rats, whereas the activities of glycolytic enzymes, body weight, levels of hemoglobin, RBC count, and packed cell volume were found to be decreased. These biochemical alterations observed in arthritic animals were ameliorated significantly after the administration of Withania somnifera root powder (1000 mg/kg/b.wt) and Indomethacin (3 mg/kg/b.wt). Our results suggest that Withania somnifera root powder is capable of rectifying the above biochemical changes in adjuvant arthritis and it may prove to be useful in treating rheumatoid arthritis

CYT387 restores Th17/Treg cell balance and inhibits mature osteoclast cell formation by regulating SOX-5 signaling in rheumatoid arthritis

The present study was conducted to investigate the therapeutic efficacy of CYT387 in rheumatoid arthritis pathogenesis concerning T helper 17 cell/regulatory T cell (Th17/Treg) imbalance and osteoclastogenesis. In this study, CYT 387 inhibited the proliferation of Th17 cells and collectively relocated FoxP3 + Treg cell differentiation in vitro. CYT387 mitigated the secretion of pathogenic IL-17 via reduced ROR-γt expression and restored the elevated expression of IL-10 by increasing FOXP3 activity. Alternatively, CYT387 abrogated RANKL expression and significantly increased osteoprotegerin levels by reducing SOX-5 activity. Furthermore, in a co-culture system of IL-6 and sIL-6R stimulated adjuvant-induced arthritic fibroblast-like synoviocytes and monocytes/macrophages collected from rat bone marrow, CYT387 reduced osteoclast formation and bone resorptive activity. In conclusion, CYT387 modulated IL-6/sIL-6R dependent JAK1/STAT3 activation and signaling mechanisms in CD4+T cells under Th17 differentiation conditions and osteoclast cells via dampened activation of SOX-5

Habbe Gule Aakh prevents glycolytic program and alleviates disease progression in a rheumatoid arthritis animal model

BackgroundThe acquired tumorigenic phenotype of the resident fibroblast like synoviocytes (FLS) is cornerstone to exacerbating rheumatoid arthritis (RA) disease progression. Toll like receptor 4 (TLR4) signalling can sustain the proliferative and invasive phenotype of these synoviocytes resulting in cartilage degradation and bone damage. A marked increase in glycolytic activity also contributes to the malignant character of these cells. Herein, we aim to study the prospects of TLR4 activation leading to improved glycolytic flux. Further, we also strategize the therapeutic modality of Habbe Gule Aakh (HGA), a polyherbal unani formulation to rescue disease progression via blockade of TLR4 activation.MethodsWe activated TLR4 signaling in SW982 cells, cultured in high glucose medium. Initially, the expression profile of glycolytic rate limiting enzymes- hexokinase 2 (HK2) and pyruvate kinase M2 (PKM2) was assessed. Next, we evaluated the ability of HGA to regulate the expression of these enzymes via ablation of TLR4 activation. Further, we investigated the pathway of glucose uptake via tissue inhibitor of matrix metalloproteinase 1 or TIMP1 and counterintuitively investigated HGA to arrest the uptake of glucose via p65-TIMP1 signaling axis. To sufficiently validate our findings, we utilised network pharmacology approach, to uncover the interactome of HGA against rheumatoid arthritis targets. Ultimately, we leveraged in-vivo models to support the anti-arthritic claims of HGA.ResultsHGA regulated the proliferation and invasive phenotype of SW982 cells cultured in high glucose medium via blockade of TLR4 activation. Further, in-silico and in-vivo approaches suggest a mechanistic insight to the anti-arthritic activity of HGA upon blockade of TLR4-mediated glycolytic flux in resident synoviocytes.ConclusionPharmacological intervention with Habbe Gule Aakh can rescue exacerbation of rheumatoid arthritis disease severity via TLR4 signaling axis. The findings of this study strengthen the rationale for the use of HGA in clinical settings involving RA patients

CYT387 restores Th17/Treg cell balance and inhibits mature osteoclast cell formation by regulating SOX-5 signaling in rheumatoid arthritis

The present study was conducted to investigate the therapeutic efficacy of CYT387 in rheumatoid arthritis pathogenesis concerning T helper 17 cell/regulatory T cell (Th17/Treg) imbalance and osteoclastogenesis. In this study, CYT 387 inhibited the proliferation of Th17 cells and collectively relocated FoxP3 + Treg cell differentiation in vitro. CYT387 mitigated the secretion of pathogenic IL-17 via reduced ROR-γt expression and restored the elevated expression of IL-10 by increasing FOXP3 activity. Alternatively, CYT387 abrogated RANKL expression and significantly increased osteoprotegerin levels by reducing SOX-5 activity. Furthermore, in a co-culture system of IL-6 and sIL-6R stimulated adjuvant-induced arthritic fibroblast-like synoviocytes and monocytes/macrophages collected from rat bone marrow, CYT387 reduced osteoclast formation and bone resorptive activity. In conclusion, CYT387 modulated IL-6/sIL-6R dependent JAK1/STAT3 activation and signaling mechanisms in CD4+T cells under Th17 differentiation conditions and osteoclast cells via dampened activation of SOX-5