Low-molecular-weight collagen peptides supplement promotes a healthy skin: A randomized, double-blinded, placebo-controlled study.

BACKGROUND: Oral collagen peptides supplementation was reported to improve skin integrity and counteract skin aging. AIMS: A randomized, double-blinded, placebo-controlled study was conducted to clinically evaluate the impact of low-molecular-weight collagen peptides on the human skin. PATIENTS/METHODS: Healthy adult participants (n = 100) were randomly assigned to receive a test product containing low-molecular-weight collagen peptides or a placebo. Parameters of skin wrinkles, elasticity, hydration, and whitening (melanin and erythema indexes) were measured at baseline and after 4, 8, and 12 weeks. RESULTS: Compared with the placebo group, the average skin roughness, maximum of all peak-to-valley values, maximum peak height of the wrinkle, and average maximum height of the wrinkle were significantly improved in the test group. Parameters of skin elasticity, including overall elasticity, net elasticity, and biological elasticity, were also significantly improved in the test group at Week 12 as compared with the placebo group. Moreover, skin hydration and whitening parameters changed more significantly in the test group than in the placebo group. None of the participants experienced adverse events related to the test product. CONCLUSIONS: Taken together, these findings suggest that low-molecular-weight collagen peptides supplementation can safely ehance human skin wrinkling, hydration, elasticity, and whitening properties.

Tolerability and efficacy assessment of an oral collagen supplement for the improvement of biophysical and ultrasonographic parameters of skin in middle eastern consumers.

BACKGROUND: Topical skin care products often do not reach the deeper layers of the skin, and oral hydrolyzed collagen is one of the newest and most popular systemic supplementations for skin rejuvenation. However, there are limited information in case of Middle Eastern consumers OBJECTIVE: The purpose of this study was to evaluate the tolerability and efficacy of an oral collagen supplement for improvement of skin elasticity, hydration, and roughness in Middle Eastern consumers. METHODS AND MATERIALS: It was a 12-week, before-after clinical study, conducted on 20 participants (18 women and 2 men) aged 44.15 ± 5.36 years with skin type III-IV. Skin elasticity parameters (R0, R2, R5, and R7), skin hydration and friction, as well as the thickness and echo density of the dermis, were measured after six and 12 weeks daily intake of the study product, as well as 4 weeks after stopping its use (week 16). Participants' satisfaction was assessed on the basis of their answers to the standard questionnaire, and tolerability of the product was assessed by monitoring the adverse effects. RESULTS: A significant improvement was detected in R2, R5, and skin friction at week 12 (p-values 0.041, 0.012 and <0.01, respectively). At week 16, the values remained at an increased level, which indicates the persistence of the results. The increase of dermis density in week 16 was also significant (p-value = 0.03). Moderate overall satisfaction was reported with the treatment, and a few gastrointestinal complications were reported. CONCLUSION: The study demonstrated that oral collagen peptides could significantly improve the skin elasticity, roughness, and dermis echo density, and they also proved to be safe and well-tolerated.

Produção de biscoitos recheados com formação de organogel na base gordurosa / Production of sandwich cookies with the formation of organogel in the fatty base

A fim de atender à demanda do público que atualmente busca por alimentos mais saudáveis, as indústrias têm procurado alternativas que possibilitem a aplicação de ingredientes que agreguem valor nutricional aos produtos. A redução de gorduras saturadas e trans em produtos alimentícios, bem como a inserção de cereais ou farinhas nutricionais, vem sendo aplicadas em produtos de panificação. Biscoitos recheados possuem como bases geralmente biscoitos à base de farinha de trigo. O objetivo foi desenvolver formulação de biscoitos recheados com substituição de gordura vegetal por organogel no recheio e de farinha de trigo por farinha de sorgo no biscoito, a fim de agregar valor nutricional ao produto. Foram desenvolvidos biscoitos recheados: 1) recheio controle e com substituição da gordura vegetal dos recheios por organogel elaborado com sistema emulsionado (colágeno + óleo vegetal + água), a fim de diminuir concentrações de gorduras saturadas e trans. 2) para a base elaborouse biscoitos controle (farinha de trigo) e com substituição parcial e total de farinha de trigo por farinha de sorgo em 50% (50FS) e 100% (100FS). Foram conduzidas nos recheios e das bases dos biscoitos análises físicas e físico-químicas (textura, atividade de água, cor, composição centesimal e reologia) para avaliação e para análise de estabilidade de 6 semanas. Os resultados apresentaram que o biscoito 50FS obteve melhor valor de textura (Controle: 16,09 ± 1,28 N; 50FS: 19,63 ± 5,68 N e 100FS: 10,09 ± 0,65 N) e menor teor de atividade de água (Semana 01: 0,327±0,01 e Semana 06: 0,389 ± 0,00) do que o biscoito controle, durante análise de estabilidade. O biscoito 100FS apresentou coloração mais avermelhada. Os biscoitos 50FS e 100FS apresentaram maior teor proteico do que o controle (Controle: 5,37 ± 0,23 %; 50FS: 5,64 ± 0,49 % e 100FS: 5,75 ± 0,49 %). O recheio com organogel apresentou maior dureza (N) durante análise de estabilidade do que o recheio controle (Semana 6 Organogel: 6,81±1,48; Controle: 4,29±0,38). Os parâmetros de adesividade, coesividade e gomosidade do recheio com organogel não apresentaram diferenças significativas (p > 0,05). Os valores de atividade de água da formulação com organogel foram mais altos do que o recheio controle (Semana 6 Organogel: 0,730±0,00; Controle: 0,555±0,01). O valor de L* foi maior para o recheio controle, apresentando coloração mais amarelada do que a formulação com organogel. O recheio com organogel apresentou redução de 65 % do teor lipídico e aumento do teor proteico. Os recheios controle, com organogel e de mercado apresentaram comportamento tixotrópico durante a avaliação reológica, sendo que o produto de mercado teve comportamento próximo à formulação controle, com recuperação quase total da estrutura. Foram desenvolvidos cinco produtos, sendo três inovadores com valor nutricional agregado, atendendo às legislações vigentes, vida útil mínima de 6 semanas e ao apelo do mercado atual, podendo ser comercializados como biscoito recheado

In order to satisfy the demand of the public that is currently looking for healthier foods industries have been looking for alternatives that allow the application of ingredients that add nutritional value to the products. The reduction of saturated and trans fats in food products, as well as the insertion of cereals or nutritional flours, has been applied in bakery products. Filled cookies are usually based on wheat flour. The objective was to develop a formulation of filled cookies with replacement of vegetable fat for organogel in the filling and wheat flour for sorghum flour in the biscuit, in order to add nutritional value to the product. In this study, cookies filled with vegetable fat and wheat flour were used as a control where: 1) filling was replaced by organogel elaborated with an emulsified system (collagen + vegetable oil + water); and 2) base was prepared with partial and total replacer of wheat flour for sorghum flour in 50% (50FS) and 100% (100FS). Physical and physicochemical analyzes (texture, water activity, color, proximate composition and rheology) were carried out on the fillings and bases of the biscuits for evaluation and for the stability analysis of 6 weeks. The results showed that the 50FS cookies had a better texture value (Control: 16,09±1,28 N; 50FS: 19,63±5,68N and 10,09±0,65 N) and lower content of water activity (Week 1: 0,327±0,01 and Week 6: 0,389±0,00) than the control cookie during stability analysis. The 100FS had a more reddish color. The 50FS and 100FS cookies had a higher protein content than the control (Control: 5,37±0,23 %; 50FS 5,64±0,49 %). The fillings with organogel showed a higher hardness (N) than the control during stability analysis (Week 6 Organogel: 6,81±1,48; Control: 4,29±0,38). The parameters of adhesiveness, cohesiveness and guminess of the filling with organogel showed no significant differences (p> 0.05). The water activity values of the organogel formulation were higher than the control filling (Week 6 Organogel: 0,730±0,00; Control: 0,555±0,01). The value of L * was higher for the control filling, showing a more yellowish color than the formulation with organogel. The filling with organogel showed a 65% reduction in lipid content and an increase in protein content. The control, organogel and market fillings showed a thixotropic behavior in the rheological evaluation, and the market product had a behavior close to the control formulation, with almost total recovery of the structure. Five products were developed, three of which were innovative with added nutritional value, in compliance with current legislation, a minimum shelf life of 6 weeks, which can be sold as a stuffed cookies.

Baicalin alleviates collagen­induced arthritis and suppresses TLR2/MYD88/NF­κB p65 signaling in rats and HFLS­RAs.

Baicalin is a flavonoid isolated from the root of Scutellaria baicalensis with anti­inflammatory, antioxidant and antiapoptotic pharmacological properties. however, the therapeutic effect of baicalin on rheumatoid arthritis (RA) is not completely understood. The present study aimed to explore the therapeutic potential and mechanisms underlying baicalin in collagen­induced arthritis (CIA) model rats. CIA was induced in male SD rats. The hind paw thickness and severity of joint injury were monitored to assess the onset of arthritis. At 28 days after the initial immunization, different doses of baicalin were administered once daily via oral gavage for 40 days. The radiologic and pathological alterations were examined using X­ray, and hematoxylin and eosin staining, respectively. ELISA was employed to measure the serum levels of proinflammatory cytokines. Reverse transcription­quantitative PCR and western blotting were conducted to determine the expression of toll­like receptor (TLR)2, myeloid differentiation factor 88 (MYD88) and NF­κB p65. Baicalin treatment noticeably alleviated radiographic and histologic abnormalities in the hind paw joints of CIA model rats in a dose­dependent manner. The serum levels of proinflammatory cytokines were significantly decreased in baicalin­treated CIA model rats compared with vehicle­treated CIA model rats. The mRNA expression levels of TLR2 and MYD88, as well as the protein expression levels of TLR2, MYD88 and NF­κB p65 were significantly decreased by baicalin treatment in the synovial tissue of CIA model rats and human RA fibroblast­like synoviocytes. The results suggested that baicalin may exert a beneficial effect on CIA, which may be mediated by inhibiting the TLR2/MYD88/NF­κB signaling pathway.

Dynamic Alterations in the Gut Microbiota of Collagen-Induced Arthritis Rats Following the Prolonged Administration of Total Glucosides of Paeony.

Rheumatoid arthritis (RA) is a common autoimmune disease linked to chronic inflammation. Dysbiosis of the gut microbiota has been proposed to contribute to the risk of RA, and a large number of researchers have investigated the gut-joint axis hypothesis using the collagen-induced arthritis (CIA) rats. However, previous studies mainly involved short-term experiments; very few used the CIA model to investigate changes in gut microbiota over time. Moreover, previous research failed to use the CIA model to carry out detailed investigations of the effects of drug treatments upon inflammation in the joints, hyperplasia of the synovium, imbalance in the ratios of Th1/Th2 and Th17/Treg cells, intestinal cytokines and the gut microbiota following long-term intervention. In the present study, we carried out a 16-week experiment to investigate changes in the gut microbiota of CIA rats, and evaluated the modulatory effect of total glucosides of paeony (TGP), an immunomodulatory agent widely used in the treatment of RA, after 12 weeks of administration. We found that taxonomic differences developed in the microbial structure between the CIA group and the Control group. Furthermore, the administration of TGP was able to correct 78% of these taxonomic differences, while also increase the relative abundance of certain forms of beneficial symbiotic bacteria. By the end of the experiment, TGP had reduced body weight, thymus index and inflammatory cell infiltration in the ankle joint of CIA rats. Furthermore, the administration of TGP had down-regulated the synovial content of VEGF and the levels of Th1 cells and Th17 cells in CIA rats, and up-regulated the levels of Th2 cells and Treg cells. The administration of TGP also inhibited the levels of intestinal cytokines, secretory immunoglobulin A (SIgA) and Interferon-γ (IFN-γ). In conclusion, the influence of TGP on dynamic changes in gut microbiota, along with the observed improvement of indicators related to CIA symptoms during 12 weeks of administration, supported the hypothesis that the microbiome may play a role in TGP-mediated therapeutic effects in CIA rats. The present study also indicated that the mechanism underlying these effects may be related to the regulation of intestinal mucosal immunity remains unknown and deserves further research attention.

Methotrexate disposition, anti-folate activity and efficacy in the collagen-induced arthritis mouse model.

Methotrexate (MTX) efficacy in autoimmune arthritis is variable and unpredictable resulting in the need for the identification of biomarkers to guide drug therapy. This study utilizes the collagen-induced arthritis mouse model to investigate erythrocyte MTX disposition and anti-folate activity as biochemical markers of efficacy in autoimmune arthritis. Following induction of arthritis, DBA/1J mice were treated with once-weekly subcutaneous MTX at varying doses over a period of 40 days. At the completion of the study tissue samples were analyzed for MTX and folate content and assessed for their relationship with MTX efficacy. MTX treatment resulted in a reduction in disease activity that was variable and dose-dependent. Erythrocyte accumulation of MTX and its polyglutamate metabolites were dose proportionate, however, polyglutamate metabolites represented a mean ±â€¯S.E.M. of 8.9 ±â€¯0.4% of total erythrocyte MTX, which is markedly lower than previously observed in humans and failed to display any significant association with MTX efficacy. MTX treatment resulted in reductions in erythrocyte 5-methyl-tetrahydrofolate (5mTHF) levels that were similar to those previously observed in human studies. Disease induction was associated with a decrease in liver 5mTHF and increased formyl-tetrahydrofolate (fTHF) that was normalized in MTX treated mice. MTX efficacy was associated with reductions in erythrocyte 5mTHF (P = 0.04) and increases in liver 5mTHF (P = 0.0001). Together, these findings demonstrate a relationship between alterations in tissue folate levels and MTX efficacy, and supports erythrocyte levels of 5mTHF as a marker of MTX efficacy in autoimmune arthritis.

Dioscin, a Steroidal Saponin Isolated from Dioscorea nipponica, Attenuates Collagen-Induced Arthritis by Inhibiting Th17 Cell Response.

Dioscin, a steroidal saponin isolated from Dioscorea nipponica Makino, has previously been shown to possess antiarthritic effects. However, the underlying mechanism is still elusive. Herein, we investigated the therapeutic effects of dioscin on collagen-induced arthritis (CIA) in DBA/1 mice and related mechanism. Cytokine production in CII-specific immune responses were measured by enzyme-linked immunosorbent assay (ELISA); Th17 cell-related gene expression, including IL-17A, ROR γτ and IL-23p19, were detected by qPCR analysis; Surface marker, T regulatory (Treg) cells and intracellular cytokines (IL-17A and IFN- γ ) were evaluated by flow cytometry. We performed Th17 cell differentiation assay in vitro. Results showed that, in vivo, dioscin treatment significantly reduced the severity of CIA, which was accompanied by decreased Th17 response, but not Th1 and Treg response; dioscin-treated mice also showed lower percentage of CD11b + Gr-1 + neutrophils; In vitro, dioscin treatment suppressed the differentiation of naive CD4 + T cells into Th17 cell and decreased IL-17A production. Collectively, our results indicate that dioscin exerts antiarthritic effects by inhibiting Th17 cell immune response.

The analgesic effect and possible mechanisms by which koumine alters type II collagen-induced arthritis in rats.

Gelsemium elegans Benth. is a toxic plant that has been used as an ancient Chinese herbal remedy for rheumatoid arthritis (RA) and nervous pain, spasticity, skin ulcers, and cancers. Koumine, one of its representative alkaloids, shows numerous promising pharmacological activities, including anti-inflammatory and analgesic activities. Here, we investigated the analgesic effect of koumine on the collagen-induced arthritis (CIA) rat model of RA and explored the potential pharmacological mechanisms underlying the analgesia. In the CIA rats, repeated koumine treatments significantly reduced pain compared to controls and attenuated the collagen-induced increase in levels of glial fibrillary acidic protein (GFAP) and the pro-inflammatory cytokines tumour necrosis factor α (TNF-α) and interleukin 1ß (IL-1ß). Cultured astrocytes showed reduced astrocyte reactivation and decreased production of both tested cytokines. Based on our results, koumine exerted both analgesic and anti-inflammatory effects on the CIA rat model that were apparently mediated by inhibiting astrocyte reactivation and pro-inflammatory cytokine production.

Diet-induced obesity and associated disorders are prevented by natural bioactive type 1 fish collagen peptides (Naticol®) treatment.

To fight against metabolic disorders such as insulin resistance, new alimentary behaviors are developed. For instance, hyperproteined, gluten-free, or collagen-enriched diets could be preconized in order to reduce the consequences of obesity. In this aim, this study evaluates the potential effects of warm sea fish collagen peptides (Naticol®) on representative metabolic and inflammatory parameters. For that, male C57Bl6/J mice fed with either a chow- (CD) or high-fat diet (HFD) were submitted or not to specific collagen peptides in drinking water (4 g/kg bw/d) for 20 weeks. Weight, body composition, glucose tolerance, and insulin sensitivity were followed up. Effects of fish collagen peptides on various blood parameters reflecting the metabolism status were also measured (free fatty acids, triglycerides, cholesterol, hormones) together with adipocyte inflammation. Results showed that HFD-fed mice supplemented by fish collagen peptides exhibited a significant lower increase in body weight as soon as the twelfth week of treatment whereas no effect of the peptide was observed in CD fed mice. In line with this result, a weaker increase in fat mass in HFD-fed mice supplemented with Naticol® at both 9 and 18 weeks of treatment was also observed. In spite of this resistance to obesity promoted by fish collagen peptides treatment, no difference in glucose tolerance was found between groups whereas mice treated with Naticol® exhibited a lower basal glycemia. Also, even if no effect of the treatment on adipocyte lipolysis was found, a decrease of inflammatory cytokines was retrieved in collagen-supplemented group arguing for a potential better insulin sensitivity. Altogether, these results need to be completed but are the first describing a benefic role of warm sea fish collagen peptides in a context of metabolic disease paving the route for a potential utilization in human obesity-associated disorders.

Hyperimmune colostrum alleviates rheumatoid arthritis in a collagen-induced arthritis murine model.

Our aging population and the accompanying decline in immune function is a growing concern that may be addressed by finding natural methods to enhance the immunocompetence of our elderly. Bovine milk and colostrum from cows that have been immunized have been shown to provide additional immunoglobulins and other bioactive molecules that enhance immune function. The purpose of this study was to investigate the ability of hyperimmune bovine colostrum to alleviate the symptoms of rheumatoid arthritis in a murine model. The collagen-induced arthritis DBA/1J murine model was used for this study. Mice were fed colostrum from immunized cows at either 5 or 10 mg/mouse per day or controls for 49 d. The data showed that the colostrum-fed groups had significantly lower total swelling scores and significantly lower collagen-specific antibody (IgG2a), inflammation-associated antibody (total IgG), and the inflammatory cytokines tumor necrosis factor α, IL-2, IL-6, and IFN-γ. The results strongly suggest that colostrum from immunized cows may have anti-inflammatory activity in a mouse model of rheumatoid arthritis.

[Effect of Moxibustion on Serum IL-17 and TNF-α Levels in Collagen-induced Arthritis Rats].

OBJECTIVE: To observe the influence of moxibustion on serum interleukin -17 (IL-17) and tumor necrosis factor alpha (TNF-α) levels in collagen-induced arthritis (CIA) rats, so as to study its mechanism underlying improvement of rheumatoid arthritis. METHODS: A total of 40 male Wistar rats were used in the present study, and 8 rats were randomly selected as a normal control group. The other 32 rats were modeled. The primary immunity emulsion was made with mixed Type Ⅱ chicken collagen and complete Freund's adjuvant, and 0.3 mL emulsion (containing 0.3 mg collagen) was injected equally into left pelma, tail root and the back. Seven days after the primary immune, the same procedure was conducted to induce the secondary immunity, and the emulsion was made with mixed Type Ⅱ chicken collagen and incomplete Freund's adjuvant. The whole course of mo-deling lasted 21 days. And then 24 CIA rats were randomly divided into model group, medication group and moxibustion group (n=8 in each group). For those of the moxibustion group, suspended moxibustion with 20 mm distance above "Zusanli"(ST 36)and "Kunlun" (BL 60) was performed for 20 min/acupoint, once daily, alternately on left and right hind limbs for 10 consecutive days. For those of the medication group, gavage of methotrexate (0.1 mg/100 g) was administrated once every 5 days, and totally two times. Left ankle joint diameter and body weight were detected, and X-ray of left tarsus was observed in each group before and after modeling or after treatment. Serum levels of IL-17 and TNF-α were determined by ELISA kits. RESULTS: After modeling, the left ankle diameter and serum concentrations of IL-17 and TNF-α increased (P<0.05), and the body weight decreased (P<0.05) in the model group compared to the control group, combined with the tarsus soft tissue swelling, joint space narrowing, bone destruction seen from the tarsal X-ray. After intervention, the ankle diameter, the serum IL-17 and TNF-α levels decreased (P<0.05), and the body weight increased (P<0.05) in both medication and moxibustion groups compared to the model group; meanwhile the tarsus soft tissue swelling and the bone deformity turned to be improved. There were no significant differences between the medication group and the moxibustion group in above mentioned indexes (P>0.05). CONCLUSIONS: Moxi-bustion is effective in CIA rats, and the mechanism may be related to the reduction of serum IL-17 and TNF-α levels.

Higher iron bioavailability of a human-like collagen iron complex.

Iron deficiency remains a public health problem around the world due to low iron intake and/or bioavailability. FeSO4, ferrous succinate, and ferrous glycinate chelate are rich in iron but have poor bioavailability. To solve the problem of iron deficiency, following previous research studies, a thiolated human-like collagen-ironcomplex supplement with a high iron content was prepared in an anaerobic workstation. In addition, cell viability tests were evaluated after conducting an MTT assay, and a quantitative analysis of the thiolated human-like collagen-iron digesta samples was performed using the SDS-PAGE method coupled with gel filtration chromatography. The iron bioavailability was assessed using Caco-2 cell monolayers and iron-deficiency anemia mice models. The results showed that (1) one mole of thiolated human-like collagen-iron possessed approximately 35.34 moles of iron; (2) thiolated human-like collagen-iron did not exhibit cytotoxity and (3) thiolated human-like collagen- iron digesta samples had higher bioavailability than other iron supplements, including FeSO4, ferrous succinate, ferrous glycine chelate and thiolated human-like collagen-Fe iron. Finally, the iron bioavailability was significantly enhanced by vitamin C. These results indicated that thiolated human-like collagen-iron is a promising iron supplement for use in the future.

Herbal formula Xian-Fang-Huo-Ming-Yin regulates differentiation of lymphocytes and production of pro-inflammatory cytokines in collagen-induced arthritis mice.

BACKGROUND: Xian-Fang-Huo-Ming-Yin (XFHM), a traditional herbal formula, has been used to treat sores and carbuncles for hundreds of years in Asia. Nowadays, its clinical effects in treatment of rheumatoid arthritis (RA) have been validated. In this study, we want to study its possible molecular mechanisms of regulating the differentiation of lymphocytes and production of pro-inflammatory cytokines in collagen-induced arthritis (CIA) mice for RA treatment. METHODS: A high performance liquid chromatography-electrospray ionization/mass spectrometer (HPLC-ESI/MSn) system was used to analyze the constituents of XFHM granules. An arthritics mouse model was induced by collagen and leflunomide (LEF) was used as a positive control medicine. Pathological changes at the metatarsophalangeal joint were studied through Safranin O and immunohistochemical staining. The differentiation of T, B and NK cells was examined by flow cytometry and pro-inflammatory cytokines were assayed using an Inflammation Antibody Array assay. The expression of key molecules of the nuclear factor κB (NF-κB) and Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathways in spleen were studied by western-blot analysis. RESULTS: In our study. 21 different dominant chemical constituents were identified in XFHM. Treatment with XFHM suppressed the pathological changes in arthrosis of CIA. Additionally, XFHM down-regulated the proliferation and differentiation of CD3+ T cells and CD3-CD19+ B cells significantly. However, XFHM had no significant effect on CD3-NK1.1+ NK cells. Further study showed that the production of pro-inflammatory cytokines had been suppressed by inhibiting the activation of NF-κB and JAK/STAT signaling. CONCLUSIONS: XFHM can regulate and maintain the immunologic balance of lymphocytic immunity and inhibit the production of pro-inflammatory cytokines, thus suppressing the pathological changes of RA. Therefore, XFHM may be used as an application of traditional medicine against RA in modern complementary and alternative therapeutics.

Harnessing the natural inhibitory domain to control TNFα Converting Enzyme (TACE) activity in vivo.

Dysregulated activity of A Disintegrin And Metalloproteinase 17 (ADAM17)/TNFα Converting Enzyme (TACE) is associated with inflammatory disorders and cancer progression by releasing regulatory membrane-tethered proteins like TNFα, IL6R and EGFR ligands. Although specific inhibition of TACE is thought to be a viable strategy for inflammatory disorders and for malignancies treatment, the generation of effective inhibitors in vivo has been proven to be challenging. Here we report on the development of a protein inhibitor that leverages the endogenous modulator of TACE. We have generated a stable form of the auto-inhibitory TACE prodomain (TPD), which specifically inhibits in vitro and cell-surface TACE, but not the related ADAM10, and effectively modulated TNFα secretion in cells. TPD significantly attenuated TACE-mediated disease models of sepsis, rheumatoid arthritis (RA) and inflammatory bowel disease (IBD), and reduced TNFα in synovial fluids from RA patients. Our results demonstrate that intervening with endogenous ADAM sheddase modulatory mechanisms holds potential as a general strategy for the design of ADAM inhibitors.

Puerarin attenuates inflammation and oxidation in mice with collagen antibody-induced arthritis via TLR4/NF-κB signaling.

Puerarin is an important active ingredient in the root of kudzu vine due to its pharmacological properties. The aim of the present study is to contribute to the existing knowledge of the effect of puerarin in the attenuation of inflammation and oxidation in mice with collagen antibody-induced arthritis via toll­like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling. Arthritis was induced using injection of anti­type II collagen antibodies. Treatment with puerarin was observed to significantly decrease clinical scoring of the collagen antibody­induced arthritis and suppress oxidative stress and the inflammatory response in mice. Furthermore, puerarin was demonstrated to inhibit mRNA expression of matrix metalloproteinase­9 and protein expression of TLR4 following collagen antibody-induced arthritis in mice. The effect of puerarin may be associated with the suppression of NF­κB activity in collagen antibody­induced arthritis mice. Furthermore, upregulation of phosphorylated (p)­Janus kinase 2 and p­signal transducer and activator of transcription 3 protein expression was suppressed by puerarin. The results of the present study indicate, for the first time, the effect of puerarin to attenuate inflammation and oxidation in mice with collagen antibody­induced arthritis via TLR4/NF-κB signaling.

Yiguanjian decoction and its ingredients inhibit angiogenesis in carbon tetrachloride-induced cirrhosis mice.

BACKGROUND: Cirrhosis is associated with angiogenesis and disruption of hepatic vascular architecture. Yiguanjian (YGJ) decoction, a prescription from traditional Chinese medicine, is widely used for treating liver diseases. We studied whether YGJ or its ingredients (iYGJ) had an anti-angiogenic effect and explored possible mechanisms underlying this process. METHODS: Cirrhosis was induced with carbon tetrachloride (CCl4) (ip) in C57BL/6 mice for 6 weeks. From week 4 to week 6, cirrhotic mice were randomly divided into four groups: sorafenib-treated, YGJ-treated and iYGJ-treated mice and placebo. Serum biochemistries, hydroxyproline (Hyp) content and histopathological changes of hepatic tissues were measured as were α-smooth muscle actin (α-SMA), collagen I, CD31, vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR) 2 and hypoxia-inducible factor (HIF)-1α. RESULTS: Both YGJ and iYGJ improved serum biochemistries. Changes of histopathology showed that YGJ and iYGJ reduced hepatic tissue necroinflammatory and collagen fiber deposition in cirrhosis mice. Compared to the CCl4 treated animals, Hyp, α-SMA, collagen I, CD31, VEGF, VEGFR, and HIF-1α expression decreased in YGJ and iYGJ groups. CONCLUSIONS: YGJ and iYGJ inhibited liver angiogenesis in cirrhotic mice treated with CCl4 by inhibiting the HIF-1α/VEGF signaling pathway, suggesting that anti-angiogenic effects of YGJ and iYGJ are associated with improving the hepatic hypoxic microenvironment.

Naringenin inhibits dendritic cell maturation and has therapeutic effects in a murine model of collagen-induced arthritis.

The aim of this study was to investigate the effect of naringenin (5,7,4'-trihydroxyflavanone), a citrus flavonoid, on dendritic cell (DC) maturation, as well as its potential as a therapeutic agent in a murine model of collagen-induced arthritis (CIA). Naringenin effectively inhibited lipopolysaccharide (LPS)-induced DC maturation as shown by reductions in the production of proinflammatory cytokines/chemokines, the expression of costimulatory molecules and the Ag-specific T cell priming ability of DCs when given at noncytotoxic doses. In addition, the decrease of LPS-induced MAPK and NF-κB signaling activation may contribute to the inhibitory activity of naringenin. In mice with CIA, the oral administration of naringenin ameliorated the severity of arthritis, reduced the levels of anticollagen Type II (CII) IgG and limited the proliferation of T cells, observed as a lower frequency of Th1 and Th17 cells in the spleen after restimulation with CII. In conclusion, this study shows for the first time that naringenin can manipulate the immunostimulatory properties of DCs and thus represents a potential therapeutic for the treatment of rheumatoid arthritis in humans.

Electroacupuncture attenuates collagen-induced arthritis in rats through vasoactive intestinal peptide signalling-dependent re-establishment of the regulatory T cell/T-helper 17 cell balance.

OBJECTIVE: Imbalance between T-helper 17 (Th17) cells and regulatory T (Treg) cells is causally linked to the development of rheumatoid arthritis (RA). In this study, we tested the hypothesis that electroacupuncture (EA) confers therapeutic benefits in RA through activation of vasoactive intestinal peptide (VIP)-dependent signalling and restoration of the Th17/Treg cell balance. MATERIALS AND METHODS: A collagen-induced arthritis (CIA) model was induced in Sprague-Dawley rats by injection of bovine type II collagen in incomplete Freund's adjuvant on day 0 and day 7. Three days after the second injection, EA was given at acupuncture points GB39 and ST36 three times per week for 4 weeks. To block VIP signalling, [D-P-Cl-Phe(6)-Leu(17)]-VIP, a VIP receptor antagonist, was administered intraperitoneally 30 min before EA. Inflammatory and pathological responses in the joint were assessed. Synovial VIP receptor mRNA levels and Treg and Th17 cell frequencies in the spleen were determined. RESULTS: EA significantly reduced the severity of CIA, as evidenced by reduced paw volumes, arthritis scores and inflammation scores. EA significantly increased mRNA expression of the VIP receptor VPAC1 and led to an elevation in CD4(+)FOXP3(+) Treg cell frequency and a reduction in CD4(+)IL17(+) Th17 cell frequency. Pre-injection of a VIP receptor antagonist significantly reversed EA-induced expansion of Treg cells, but did not alter the frequencies of Th17 cells. CONCLUSIONS: EA exerts anti-inflammatory effects in a collagen-induced rat model of arthritis. These effects appear to be mediated through activation of VIP signalling and re-establishment of the Th17/Treg cell balance.

Anti-thrombotic effect of rutin isolated from Dendropanax morbifera Leveille.

Dendropanax morbifera H. Lev. is well known in Korean traditional medicine for improvement of blood circulation. In this study, rutin, a bioflavonoid having anti-thrombotic and anticoagulant activities was isolated from a traditional medicinal plant, D. morbifera H. Lev. The chemical characteristics of rutin was studied to be quercetin 3-O-α-l-rhamnopyranosyl-(1-6)-ß-d-glucopyranoside using high performance liquid chromatography mass spectrometry (HPLC-MS), proton nuclear magnetic resonance ((1)H NMR) and carbon-13 nuclear magnetic resonance ((13)C NMR). Turbidity and fibrin clotting studies revealed that rutin reduces fibrin clot in concentration dependent manner. Rutin was found to prolong activated partial thromboplastin time (aPTT), prothrombin time (PT) and closure time (CT). Furthermore, it decreased the activity of pro-coagulant protein, thrombin. In vivo study showed that rutin exerted a significant protective effect against collagen and epinephrine (or thrombin) induced acute thromboembolism in mice. These results suggest that rutin has a potent to be an anti-thrombotic agent for cardiovascular diseases.

Therapeutic effect of Cryptotanshinone on collagen-induced arthritis in rats via inhibiting nuclear factor kappa B signaling pathway.

The discovery of new therapeutic drugs with the ability of preventing inflammation and joint destruction with less adverse effects is extremely urgent for rheumatoid arthritis (RA). Cryptotanshinone (CTS), an active component isolated from the root of Salvia miltiorrhiza Bunge, has been reported to have antibacterial and antitumor effects. However, its effects on RA have not been clearly elucidated. Here, we investigated the therapeutic effect of CTS on type II collagen-induced arthritis (CIA) in rats and explored the underlying mechanisms. Our results revealed that CTS treatment efficaciously ameliorated inflammation and joint destruction of rats with CIA. Both in vivo and in vitro studies showed that CTS suppressed the production of proinflammatory cytokines including interleukin 1ß, tumor necrosis factor alpha, and interleukin 17α production and downregulated the production and activity of matrix metalloproteinase 9. By receptor activator of nuclear factor kappa B (NF-κB) ligand-induced bone marrow macrophages, we observed that CTS could inhibit osteoclast differentiation, which is critic for joint destruction. Further studies on inflammatory signaling revealed that CTS could inhibit the degradation of inhibitor of NF-κB (IκB) α in vivo and in vitro, prevent the nuclear translocation of NF-κB p65 induced by lipopolysaccharide in a time- and dose-dependent manner. By electrophoretic mobility shift assay and luciferase reporter assay, we found that CTS distinctively inhibited the NF-κB DNA binding activity and NF-κB-dependent luciferase activity. These results indicate that the therapeutic effect of CTS on CIA is accomplished mainly through the inhibition of NF-κB signaling. Our findings provide the evidence to develop CTS as a potential therapeutic agent for patients with RA.