withanolides and Arthritis--Rheumatoid

Rheumatoid arthritis (RA) is an inflammatory autoimmune disease that affects the synovial joints. Nearly 1.6 billion patients are affected by RA worldwide and the incidence of RA is about 0.5 to 1%. Recent studies reveal that immune cell responses and secretion of inflammatory factors are important for the control of RA.. In this study, a set of 402 phytochemicals with anti-inflammatory properties and 16 target proteins related to anti-inflammatory diseases were identified from the literature and they were subjected to network analysis. The protein-protein interaction (PPI) network was constructed using STRING (Search Tool for the Retrieval of Interacting Genes database) database. Visualization of the target gene-phytochemical network and its protein-protein interaction network was conducted using Cytoscape and further analyzed using MCODE (Molecular Complex Detection). The gene ontology and KEGG pathway analysis was performed using DAVID tool.. Our results from the network approach indicate that the phytochemicals such as Withanolide, Diosgenin, and Butulin could act as potential substitute for anti-inflammatory drugs, including DMARDs. Genes such as Mitogen-activated protein kinase (MAPK) and Interleukin were found as hub genes and acted as best inhibitors for the target protein pathways. Curcumin, Catechin was also found to be involved in various signaling pathways such as NF-kappa B signaling pathway, ErbB signaling pathway and acted as the best inhibitor along with other candidate phytochemicals.. In the current study, we were able to identify Withanolide, Diosgenin, and Butulin as potential anti-inflammatory phytochemicals and determine their association with key pathways involved in RA through network analysis. We hypothesized that natural compounds could significantly contribute to the reduction of dosage, improve the treatment and act as a therapeutic agent for more economical and safer treatment of RA.

Topics: Anti-Inflammatory Agents; Arthritis, Rheumatoid; Diosgenin; Humans; Phytochemicals; Withanolides

In order to develop a better therapeutic approach for the treatment of rheumatoid arthritis (RA), withaferin-A; a steroidal lactone incorporated with mannosylated liposomes (ML-WA) was administered to adjuvant induced arthritic rats in intent to target the synovial macrophages. The confocal microscopy studies showed a successful internalization of ML-WA in the primarily isolated synovial macrophages. Consequently, targeting synovial macrophages via ML-WA reduced the oxidative stress (ROS and NO), and paw edema, however, a progressive gain in the body weight was observed in AIA rats. ML-WA treatment upregulated the production of osteoprotegerin (OPG) and downregulated the release of receptor activator of nuclear factor-κB ligand (RANKL), favoring osteoclastogenesis negatively. Correspondingly, the ankle joints were found intact with no bone erosion and cartilage degradation in ML-WA treated AIA rats as evidenced by histopathological analysis. Also, synovial macrophage assessment showed that the concentration and the gene amplification of M1 macrophage mediated pro-inflammatory mediators (TNF-α, IL-1β, IL-6, MCP-1 and VEGF) were curtailed in ML-WA treated AIA rats. In contrast, anti-inflammatory cytokine (IL-10) was found abundantly released. Furthermore, the mRNA expression of the M1 surface marker (CD86) was found down regulated, whereas, M2 marker (CD163) was highly amplified in ML-WA treated synovial macrophages of arthritic rats. Cumulatively, our result signified that targeted delivery of ML-WA ameliorated the severity of inflammation and bone resorption in AIA rats via M1 to M2 macrophage repolarization.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Cytokines; Female; Gene Expression; Inflammation Mediators; Lactones; Liposomes; Macrophage Activation; Macrophages; Male; Mannose; Microscopy, Confocal; Microscopy, Electron, Scanning; Rats, Wistar; Withanolides

Withania somnifera (WS) is an important herb with known antiinflammatory activity. Its molecular mechanism of action has not been investigated. The effect of a WS crude ethanol extract was studied on peripheral blood mononuclear cells of normal individuals and rheumatoid arthritis (RA) patients and synovial fluid mononuclear cells of RA patients in vitro. The WS extract significantly suppressed lipopolysaccharide (LPS) induced production of proinflammatory cytokines TNF-alpha, IL-1beta and IL-12p40 in normal individuals and RA patients, but had no effect on IL-6 production at the protein and transcript level. WS also suppressed LPS activated nitric oxide production in the mouse macrophage cell line, RAW 264.7. The extract inhibited nuclear translocation of the transcription factors NF-kappaB and AP-1 and phosphorylation of IkappaBalpha in normal and RA patients' mononuclear cells. HPLC analysis of the crude extract showed the presence of withaferin A and pure withaferin A also inhibited NF-kappaB translocation. The study demonstrated that the WS crude ethanol extract suppressed the production of proinflammatory molecules in vitro. This activity is partly through the inhibition of transcription factors NF-kappaB and AP-1 by the constituent withanolide. The role of additional constituents needs to be studied. Studies on the mechanism of action of the extract may yield potentially useful compounds for the treatment of inflammatory diseases.

Topics: Animals; Arthritis, Rheumatoid; Cell Line; Chromatography, High Pressure Liquid; Cytokines; Ergosterol; Humans; I-kappa B Proteins; Leukocytes, Mononuclear; Lipopolysaccharides; Mice; NF-kappa B; NF-KappaB Inhibitor alpha; Nitric Oxide; Phosphorylation; Plant Extracts; RNA, Messenger; Synovial Fluid; Transcription Factor AP-1; Withania; Withanolides