Muscadine Grape and Wine Polyphenols Alleviated the Severity of Collagen Induced Arthritis in Mice

Objectives The aim of this study was to investigate the effect of concentrated muscadine grape polyphenols (MGP) and muscadine wine polyphenols (MWP) on the onset and progression of collagen induced arthritis (CIA) in mice. Methods MGP and MWP were concentrated using Amberlite XAD16N resin. Polyphenol composition was determined on HPLC. Male DBA/1 mice were divided into four treatment groups as (1) healthy control, (2) CIA control, (3) CIA + MWP, and (4) CIA + MGP. Arthritis was induced by intradermal injection of type II collagen on days 0 and day 21. Mice in groups 3 and 4 were gavaged with MWP and MGP using a dose of 400 mg/kg body weight for a total of 21 days. Severity of CIA was evaluated using clinical arthritic score and inflammation in the hind-paws. Plasma levels of cytokines, proteases, and anti-collagen antibody were measured using ELISA. Results MGP and MWP contained anthocyanin 3, 5-diglucosides, flavonols, and ellagic acid. Mice in the CIA control group had an onset of arthritis on day 18, which was delayed to day 22 and 24 by MWP and MGP, respectively (P < 0.05). Severity of arthritis was much lower in mice gavaged with muscadine polyphenols after the onset. On day 42, the average arthritic score of mice in the CIA control group progressed to 7.75 on a scale of 0–16, while MGP and MWP significantly reduced this score to 3.75 and 4.00, respectively (P < 0.01). In addition, MGP and MWP significantly reduced the plasma concentration of TNF-α, IL-6, anti-collagen antibodies, and matrix metalloproteinase-3 in CIA mice but not to the same level of healthy mice. The plasma concentration of IL-17 was drastically elevated in CIA mice, but it was not affected by MWP or MGP like other cytokines. This suggested that muscadine polyphenols had limited impact on the Th17 cells in the immune system. Mice gavaged with MPG and MWP had comparable plasma cytokine content, suggesting similar anti-inflammation activity. Conclusions Muscadine grape and wine polyphenols blunt the development of arthritis in mice by reducing the levels of key inflammatory cytokines, antibodies, and proteases, and may offer a promising dietary approach to manage arthritic symptoms. Funding Sources Florida Department of Agriculture and Consumer Service and Florida Viticulture Advisory Council.

Effect of ethanol extract of Punica granatum L against Freund’s complete adjuvant-induced arthritis in rats

Purpose: To investigate the protective effect of ethanol extract of P. granatum against arthritis in rat model. Methods: Twenty-six adult male Wistar rats (120 - 150 g) were separated into four groups (n = 6): normal control, arthritic control and two treatment groups. With the exception of normal control group, arthritis was induced by intraplantar administration of Freund’s complete adjuvant (FCA) on the 1st day of drug administration. The arthritic control group was not treated, while the treatment groups received extract orally at 500 or 750 mg/kg for the period of 4 weeks and at the end of each week, paw volume, thermal hyperalgesia, arthritic score and mechanical nociceptive threshold were performed to assess arthritis. Biochemical indicators and inflammatory cytokines in serum were determined using standard procedures. Results: There was significant decrease in paw volume and arthritic score; paw withdrawal latency was enhanced in extract-treated groups, compared to arthritic control group (p < 0.05). Furthermore, ALT, AST and ALP levels, as well as RF and MDA activities decreased significantly with extract treatment, compared with arthritic control group (p < 0.05). Treatment with the extract attenuated the altered level of interleukin 1β (IL-1β) and TNF-α levels in arthritic rats. Histological examination showed that treatment with the extract significantly reversed histological changes induced by arthritis. Conclusion: The results reveal that the beneficial effect of ethanol extract of P. granatum against FCAinduced arthritis is due to its ability to reduce the levels of inflammatory cytokines.

Anti-Inflammatory and Anti-Arthritic Potential of Standardized Extract of Clerodendrum serratum (L.) Moon

Aims: Scientific biological evaluation of standardized extracts is becoming one of the central needs for the globalization of customary medication in current times. And to validate the presence of active constituents in crude medicinal extracts, analytical techniques like HPLC and HPTLC are the most suitable authentication systems. In the current study we aimed to standardize and evaluate Clerodendrum serratum (L.) Moon (Verbenaceae). For its unique anti-inflammatory and anti-arthritic properties. Evaluation and analysis of the plant, therefore, offers a new platform for the development of the herbal drug and could prove to be a safe and cost effective treatment for arthritis management.Methods: The aqueous extract of C. serratum, a common plant in the Southeastern Asian region, was used for phytochemical investigation and standardization by HPTLC and HPLC. The standardized HPLC method was further validated by using ICH guidelines. The standardized extract was investigated for anti-inflammatory and anti-arthritic activity. Complete Freund’s adjuvant (CFA) model was performed to evaluate the activity. Paw diameter, joint diameter, arthritic score, and body weight was accepted as a parameter for the evaluation of biological activity.Results: HPTLC method revealed the presence of ursolic acid with an Rf value of 0.38 and the amount quantified was 0.03% w/w. The presence of the bioactive phytochemical was further analyzed and confirmed by HPLC for which the validation was done successfully in accordance with ICH guidelines. The assay content for ursolic acid was found to be 0.059% with relative standard deviation (RSD) &lt;2.5% for specificity and precision with spike recovery between 95–110%. The anti-arthritic activity of aqueous extract exhibited COX-2 and TNF-α inhibition as observed in various parameters like paw edema, arthritic index, and joint diameter. Plant extract showed reclamation of arthritis in regard to body weight, arthritic score, paw edema, and joint diameter. The extract showed significant results for TNF-α and COX-2(p &lt; 0.0001). The plant extract also exhibited in-vitro anti-inflammatory activity.Conclusion: The current study established the scientific basis of ethnomedicinal use of the plant for anti-inflammatory purposes and the management of arthritis and can also be used for quality control purposes.

The possible role of heat shock protein-70 induction in collagen-induced arthritis in rats

AimThis study aimed to evaluate the possible role of heat shock protein-70 (HSP70) induction by 17-allylaminodemethoxygeldanamycin (17-AAG) in collagen-induced arthritis in rats.Material and methodsMale Wistar rats were divided into five groups (n = 10/group) and were treated intraperitoneally twice a week for 4 weeks, namely normal control (saline), arthritis control (AR; saline), AR + 17-AAG, AR + methotrexate (MTX), and AR + 17-AAG + MTX. At the end of the treatments, arthritic score was determined and then the animals were sacrificed. Erythrocyte sedimentation rate (ESR), serum levels of HSP70, interleukin-17 (IL-17), tumor necrosis factor-alpha (TNF-α), rheumatic factor (RF), C-reactive protein (CRP), malondialdehyde (MDA), glutathione peroxidase (GPx), and matrix metalloproteinase-9 (MMP-9) were determined.ResultsIn the AR group, all parameters increased significantly, except for GPx, which showed a pronounced decrease. The 17-AAG and/or MTX treatments significantly reduced arthritic score, ESR, IL-17, TNF-α, RF, CRP, MDA, and MMP-9 with significant increase in GPx compared to the AR group. The HSP70 level was significantly higher in the AR + 17-AAG and the AR + 17-AAG + MTX groups but significantly lower in the AR + MTX group as compared to the AR group. Also, it was significantly lower in the AR + MTX group as compared to the AR + 17-AAG group.ConclusionWe concluded that HSP70 induction by 17-AAG attenuated the inflammatory process in a rheumatoid arthritis (RA) model induced by collagen, which suggested that HSP70 inducers can be promising agents in the treatment of RA.

Inhibition of Oxidative Stress in Brain During Rat Adjuvant Arthritis by Carnosine, Trolox and Novel Trolox-Carnosine

Carnosine (CARN) is an anti-glycating agent able to quench superoxide, and to neutralize 4-hydroxynonenal. Trolox-carnosine (CARN-T) was synthesized because of its resistance against degradation and to improve CARN antioxidant capacity. We evaluated the impact of trolox (TRO), CARN and its derivative CARN-T on oxidative stress (OS) in brain during rat adjuvant arthritis (AA). The experiments were done on healthy, control arthritic and arthritic animals with administration of CARN 150 mg/kg b.w., TRO 41 mg/kg b.w. and CARN-T 75 mg/kg b.w. in a daily dose during 28 days. Antioxidants did not affect the body weight on day 14, but on day 28 TRO enhanced the weight reduction. On day 14 and 28 CARN-T and TRO reduced arthritic score. IL-1beta, MCP-1 and MMP-9 were measured in plasma on day 14. MCP-1 was decreased by CARN-T and TRO. All antioxidants reduced IL-1beta and MMP-9 levels. Malondialdehyde, 4-hydroxynonenal and protein carbonyls were increased in brain. CARN, CARN-T and TRO prevented higher lipid and protein oxidation in brain. CARN and CARN-T caused no weight reduction like TRO that has an advantage in inflammatory arthritis. Moreover the antioxidants administered had a similar therapeutic effects on arthritic score, markers of inflammation in plasma and OS in brain.

Rheumatoid Arthritis as a New Disease-Target for Bortezomib.

Bortezomib (Velcade) is the first proteasome inhibitor that has been introduced into the clinic for the treatment of multiple myeloma. Bortezomib, by inhibiting the NFkB function, results in increased tumor cell sensitivity to chemotherapy and induces apoptosis of alloreactive T-cells. Since NF-kB activation results also in transcription of pro-inflammatory molecules, it is speculated that NF-kB inhibition may also ameliorate inflammatory and autoimmune conditions. Rheumatoid arthritis may be a possible target disease for proteasome inhibitors because the most important pro-inflammatory mediators (TNF-a, IL-1, IL-6, V-CAM-1) are regulated by NF-kB. We investigated the effect of Bortezomib in a rat model of Adjuvant Arthritis (AA) which closely resembles human rheumatoid arthritis. The presence of 5,10,100 nM bortezomib for 72h or the presence of 10 nM bortezomib for 48 and 72 h in culture with ConA–activated T-cells from the spleen of rats after AA induction, significanlty inhibited their proliferation as measured by radioactive thymidine incorporation (p&lt;0,00005 in all concentrations and culture times tested). Likewise, the presence of 10 nM Bortezomib for 48 and 72 h in fibroblast-like synoviocytes (FLS) culture from rats after AA induction, significanlty inhibited proliferation (p&lt;0,00005). Bortezomib’s effect in AA was also explored in vivo by intraperitoneal injections at 0,25mg/kg×2days×2weeks in rats after AA induction by FCA injection in the tail base. Adjuvant artrhitis usually starts to develop at day 11–13 post induction. Bortezomib was administered before the onset of arthritis at the onset of arthritis and in established arthritis. The total arthritic index represented a score based on redness, swelling and deformity of the joint. Arthritis grading was assessed on a 4-point scale per limb with a maximum arthritic score of 16 per rat. The histologic severity of arthritis was also assessed by a histopathological score based on the presence of synovial hyperplasia, inflammatory infiltration, chondroplasia, bone destruction and pannus formation (max score 12 per animal). A significant delay in arthritis onset accompanied by a milder clinical picture was observed in the group which received Bortezomib before the onset of arthritis (mean day21 arthritic score; 1). More importantly, there was a dramatic reduction in the disease score in the group treated with Bortezomib both at the onset of arthritis (mean day26 arthritic score;4,1) and at advanced disease status (mean day30 arthritic score; 3,3) as compared to the untreated AA control group (mean day21, day26 and day30 arthritic score 9,6, 9,9 and 11,3 respectively/p=0,03, p=0,02 and p=0,02, respectively). There was a correlation between the clinical and histopathological assessment of disease severity in Bortezomib-treated animals vs control rats (histological score 3,75 vs 9,8 respectively). A marked reduction in the number of inflammatory cells with little or no bone erosion and limited pannus formation could be seen in the joints of bortezomib-treated rats. A significant reduction of CD3+ cells in the joints and inhibition of NF-kB in spleen sections of Bortezomib-treated rats was detected by immunohistochemistry. Flow cytometry analysis demonstrated a significant increase of CD4+ cells with reduction of CD8+, CD11b+ cells in the blood (p=0,003 and p=0,0002, respectively) and the spleen (p=0,01 and p=0,017, respectively). T-cells from Bortezomib-treated animals exhibited a higher degree of apoptosis measured as Annexin-V + cells, when compared to T-cells from AA control rats. In CT scans before and after treatment with Bortezomib of animals that received bortezomib in advanced disease phase, a significant reduction of soft tissue swelling around the affected joints with restoration of focal osteopenia and bone erosion could be detected. Currently, we are testing the combination of Bortezomib with Mesenchymal Stem Cells in the same disease model, as bortezomib has been shown to enhance the osteogenic potential of MSCs. Our data suggest that Bortezomib exhibits significant anti-inflammatory and proapoptotic activity in a rat model of adjuvant arthritis and human rheumatoid arthritis may represent an important clinical opportunity for this drug