IMMUNOLOGICAL STUDY ON ACUPUNCTURE TREATMENT OF COLLAGEN-INDUCED ARTHRITIS IN CX43-KNOCK-OUT RATS

Objective To observe the therapeutic effect of acupuncture for collagen-induced arthritis （CIA） in Cx43 knock-out mice and its underlying immunological mechanism.Methods Heterozygote （Cx43^＋/-） mice （n=64） and wild-type （Cx43^＋/＋） mice （n=46） were used in the present study.CIA model was es- tablished by intracutaneous injection of bovine collagenⅡ（immunonization）.Acupuncture of Zúsānlǐ（足三里ST36） was carried out from the 4^th week on after the initial immunization,once a day for 3 weeks.Arthritis score and days of onset of CIA were recorded.Intracellular contents of Th subgroups of splenic lymphocytes were detected with flow cytometry.Results The incidence rate of CIA and arthritis score in Cx43^＋/- mice were significantly lower than those in Cx43^＋/＋ mice （P〈0.05）.In Cx43^＋/＋ mice,3 weeks after acupuncture treatment,the score of arthritis symptoms and signs in acupuncture group was significantly lower than that in model group （P〈0.01）,while in Cx43^＋/- mice,no significant difference was found between model group and acupuncture group in arthritis score （P〉0.05）.In comparison with control group,percentages of both Thl and Th2 and Th1/Th2 increased significantly in 0x43^＋/＋ CIA model group （P〈0.05）,while compared with model group,the percentage of Th1,Th2 and Th1/Th2 in acupuncture group decreased evidently （P〈0.05） or moderately,showing that acupuncture can effectively suppress arthritis-induced increase of Thl in Cx43^＋/＋ mice.In 0x43^＋/- mice,Th1 level and Th1/Th2 of CIA model group increased considerably in comparison with control group （P〈0.05）,while compared with model group,the Th1,Th2 and Th1/Th2 of acupuncture group had no significant changes （P〉0.05）,indicating that the effect of acupuncture in suppressing increase of Th1 percent and Th1/Th2 was eliminated.Conclusion Acupuncture of Zúsānlǐ（足三里ST 36） can effectively relieve CIA symptoms and signs and inhibit increase of splenic Th1 in Cx43^＋/＋ mice but has no any effect on CIA score,Th1,Th2 and Th1/Th2 in Cx43^＋/- mice.It shows the effect of acupuncture in improving CIA is closely related to Cx43 gene.

Construction and validation of a risk prediction model for depressive symptoms in a middle-aged and elderly arthritis population

BACKGROUND Arthritis is a prevalent and debilitating condition that affects a significant proportion of middle-aged and older adults worldwide.Characterized by chronic pain,inflammation,and joint dysfunction,arthritis can severely impact physical function,quality of life,and mental health.The overall burden of arthritis is further compounded in this population due to its frequent association with depression.As the global population both the prevalence and severity of arthritis are anticipated to increase.AIM To investigate depressive symptoms in the middle-aged and elderly arthritic population in China,a risk prediction model was constructed,and its effectiveness was validated.METHODS Using the China Health and Retirement Longitudinal Study 2018 data on middleaged and elderly arthritic individuals,the population was randomly divided into a training set(n=4349)and a validation set(n=1862)at a 7:3 ratio.Based on 10-fold cross-validation,least absolute shrinkage and selection regression was used to screen the model for the best predictor variables.Logistic regression was used to construct the nomogram model.Subject receiver operating characteristic and calibration curves were used to determine model differentiation and accuracy.Decision curve analysis was used to assess the net clinical benefit.RESULTS The prevalence of depressive symptoms in the middle-aged and elderly arthritis population in China was 47.1%,multifactorial logistic regression analyses revealed that gender,age,number of chronic diseases,number of pain sites,nighttime sleep time,education,audiological status,health status,and place of residence were all predictors of depressive symptoms.The area under the curve values for the training and validation sets were 0.740(95%confidence interval:0.726-0.755)and 0.731(95%confidence interval:0.709-0.754),respectively,indicating good model differentiation.The calibration curves demonstrated good prediction accuracy,and the decision curve analysis curves demonstrated good clinical utility.CONCLUSION The risk prediction model developed in this study has strong predictive performance and is useful for screening and assessing depression symptoms in middle-aged and elderly arthritis patients.

Effectiveness of Huai Qi Huang Granules on Juvenile Collagen-induced Arthritis and Its Influence on Pyroptosis Pathway in Synovial Tissue

Summary:Huai Qi Huang(HQH)exerts great effects in clinic,such as anti-inflammation,immune-regulation,anti-cancer,and so on.However,the mechanism by which HQH protects juvenile idiopathic arthritis(JIA)is obscure.Thus,we explored deeply the protective mechanisms in juvenile collagen-induced arthritis(CIA)rat model.Pyroptosis is Gasdermin D(GSDMD)-dependent programmed cell death,involved in many diseases,such as sepsis.We investigated whether GSDMD-induced pyroptosis take part in mechanisms of juvenile CIA arthritis.Juvenile Wistar rats(3-4 weeks)were injected intradermally with fully emulsified bovine typeⅡcollagen and complete Freund's adjuvant to establish CIA rat models.Later,the CIA rats received oral administration of HQH(4.16 g/kg)once a day from the day 21 of modeling,with the treatment lasting for 28 days.Varieties of indicators were measured for evaluation of anti-inflammation effect of HQH,including hind paw swelling,arthritis scores,micro CT,and histopathological changes and the level of pro-inflammatory cytokines in the serum,including tumor necrosis factor alpha(TNF-α)and interleukin-18(IL-18).The expression of GSDMD and caspasein the joint synovial tissues was detected.The results demonstrated that the expression of the pyroptotic protein GSDMD and its upstream caspase-1 was significantly increased in the synovial tissues of CIA rats.The treatment of HQH ameliorated the symptoms in CIA rats,reduced levels of pro-inflammatory cytokines and hind paw swelling,down-regulated the expression of GDSMD and caspase-1.GSDMDinduced pyroptosis participated in the pathogenesis of CIA rats.The study supported that HQH can effectively improve joints inflammation of juvenile collagen-induced arthritis rats by inhibiting pyroptosis pathway in the joint synovial tissues.

Effects of Triptolide on the Expression and Activity of NF-κB in Synovium of Collagen-induced Arthritis Rats

Summary： The expression and activity of NF-kB in the synovium of collagen-induced arthritis （CIA） rats was detected in order to investigate the possible therapeutic effects of triptolide on rheumatoid arthritis （RA）. The experimental Wistar rat model of CIA was set up by intradermal injection of emulsion of bovine collagen 11 and the successful rate of setting-up models was evaluated by arthritis index （AI）. Rats were grouped randomly into three groups： normal, model and treatment group. The expression of TNF-α and IL-6 in synovial fluid was detected by ELISA, and the expression and activity of NF kB in synovium by immunohistochemistry method and by electrophoretic mobility shift assay （EMSA） respectively. As compared with normal group, the expression of TNF a and IL-6 in synovia （P〈0. 05）, and the expression and activity of NF-kB （P〈0.05） in synovium were increased in model group. There was statistical difference in above-mentioned indexes between model group and treatment group. Triptolide may play a protective role in IRA via downregulating the expression and activity of NF-kB in synovium.

Prediction of Response of Collagen-induced Arthritis Rats to Methotrexate： An ~1H-NMR-based Urine Metabolomic Analysis

Over one half the patients with rheumatoid arthritis （RA） are being treated with methotrexate （MTX）. Although well proven, the efficacy of MTX varies in individual patients. This study examined the metabolic biomarkers that can be used to predict the therapeutic effect of MTX by using metabolomic analysis. Rats were immunized with collagen to rapidly cause collagen-induced arthritis （CIA） and then treated with 0.1 mg/kg MTX for 4 weeks. The clinical signs and the histopathological features of CIA were observed to evaluate the therapeutic effects. Urine samples of CIA rats were collected, and analyzed by using 600 M 1H-nuclear magnetic resonance （1H-NMR） for spectral binning after the therapy. The urine spectra were divided into spectral bins, and 20 endogenous metabolites were assigned by Chenomx Suite. Multivariate analyses were performed to identify the spectral pattern of endogenous metabolites related to MTX therapy. The results showed that the clustering of the spectra of the urine samples from the responsive rats （n=20） was different from that from the non-responsive rats （n=11）. Multivariate analysis showed difference in metabolic profiles between the responsive and non-responsive rats by using partial least squares-discrimination analysis （PLS-DA） （R2=0.812, Q2=0.604）. In targeted profiling, 13 endogenous metabolites （uric acid, taurine, histidine, methionine, glycine, etc.） were selected as putative biomarkers for predicting therapeutic response to MTX. It was suggested that 1H-NMR-based metabolomic analysis can be used to predict the therapeutic effect of MTX, and several metabolites were found to be related to the therapeutic effects of MTX.

Therapeutic Actions of the Chinese Herbal Formulae with Cold and Heat Properties and Their Effects on Ultrastructures of Synoviocytes in Rats of the Collagen-Induced Arthritis

The therapeutic actions of Qing Luo Yin (QLY清络饮) with heat property and Wen Luo Yin (WLY温络饮) with cold property on pain, swelling of the ankle, arthritis index and ultrastructures of synoviocytes were compared in rats of type II collagen-induced arthritis (CIA), with tripterygium glycosidorum (TG) used as control. The results indicated that both QLY and WLY could reduce pain, swelling of the ankle and the arthritis index of CIA, and QLY had better effects in reducing the swelling of the ankle and controlling the secondary pathological lesions as compared with WLY. Investigation on the ultrastructures of synoviocytes indicated that both QLY and WLY could reduce the number of Golgi apparatus, rough surface endoplasmic reticulum, dense bodies, matrix filaments and vacuoles so as to suppress the excessive secretion of synoviocytes in rats of CIA.

Expression of Peptidylarginine Deiminase 4 and Protein Tyrosine Phosphatase Nonreceptor Type 22 in the Synovium of Collagen-Induced Arthritis Rats

Objective To study the expression level of peptidylarginine deiminase 4(PADI4) and protein tyrosine phosphatase nonreceptor type 22(PTPN22) in the synovium of rat model of collagen-induced arthritis, and to explore their possible therapeutic role in rheumatoid arthritis. Methods Thirty-two female Wistar rats weighing 100±20 g were randomly assigned into 3-week collagen-induced arthritis(CIA) model group(n=8), 4-week CIA model group(n=8), 6-week CIA model group(n=8), and the control group(n=8). The body weight changes of each group were recorded. The expression levels of PADI4 and PTPN22 were detected and compared by the methods of immunohistochemical staining and Western blot. Results Arthritis of rat began to form 14 days after sensitization and the joint swelling reached peak at 28 days. The weights of the rats slowly grew both in CIA model groups and the control group. Immunohistochemical staining results showed that the positive expression of PADI4 and PTPN22 was mainly located in cartilage peripheral mononuclear cells, the cytoplasm of infiltrated cells, and bone marrow cavity. There were significant differences in the optical density of PADI4 and PTPN22 among CIA model groups and the control group(PADI4, 0.2898±0.012, 0.2982±0.022, 0.2974±0.031, 0.2530±0.013 in 3-week CIA model, 4-week CIA model, 6-week CIA model and control groups; PTPN22, 0.2723±0.004, 0.2781±0.010, 0.2767±0.008, 0.2422±0.019; all P <0.05). The expression bands of PADI4 were observed in Western blot 3 weeks after initial immunization, the thickest in the 4th week, and decreased in the 6th week. The expression bands of PTPN2 were observed at all the time points, with no obvious time-dependent trend. Conclusions PADI4 and PTPN22 are obviously correlated with CIA in rat model. PADI4 is expressed at early stage of the disease, while the expression of PTPN22 sustains throughout the course.

Comparison of Three Experimental Models for Rat Osteoarthritis Induction

Background: Osteoarthritis is a slowly progressive and debilitating disease with high prevalence in adult population. Knee is one of the joints most affected by this disorder. There are several models for animals’ osteoarthritis induction, however it is not identified any paper that compares these techniques. The present study was aimed to define the most appropriate model for rats osteoarthritis induction. Material and Methods: 40 Wistar rats were distributed into 4 groups of 10 animals each: normality group (NG);meniscectomy group (MG);quinolone group (QG) and iodoacetate group (IG). Radiographic images of the rat’s knees were analyzed as well as the amount of chondrocytes in the epiphyseal and articular cartilage. Results: In the radiographic analysis, there was a low correlation between the raters. Regarding the amount of chondrocytes in the epiphyseal cartilage, it was noticed that the IG and QG groups had fewer chondrocytes than NG, in contrast to MG that reported similar results to normality (p > 0.05). There was no significant difference between IG and QG groups (p > 0.05). Regarding the amount of chondrocytes in articular cartilage, it was noticed that the IG group showed fewer chondrocytes than NG (p 0.05). There was no significant difference between QG and MG groups (p > 0.05). Conclusion: Intraarticular injection of iodoacetate in rats is the model with greatest effect on reduction of chondrocytes amount.

Preparation and analysis of active rat model of rheumatoid arthritis with features of TCM toxic heat-stasis painful obstruction

Objective:To establish a collagen type II-induced rat model of rheumatoid arthritis(RA)presenting characteristics of the human form of the traditional Chinese syndrome pattern of toxic heat-stasis painful obstruction(bi zheng;arthromyodynia)as well as pathologic features of active RA.The Chinese herbal medicine Tengmei decoction was used to validate the animal model.Methods:Ninety specific pathogen free Sprague-Dawley rats were randomly divided into a normal group of 6 rats and a model group of 84 rats.To establish the rat model of collageninduced arthritis(CIA),bovine type II collagen in complete Freund’s adjuvant was injected into the model group rats as a priming dose(Day 0)and boosting dose(Day 9).Changes in arthritic index(AI)scores,including limb swelling,were monitored.Thereafter,24 successfullyestablished CIA rats were randomly assigned to 4 groups with 6 animals each:model,positive control drug,high-dose traditional Chinese herbal medicine,and traditional Chinese herbal medicine.A blank control group of 6 rats was included.After 12 weeks of intervention with Tengmei decoction,articular synovial tissue and serum specimens were collected to detect interleukin-2(IL-2)and IL-17 transcription and protein expression levels.

Systematic review of robust experimental models of rheumatoid arthritis for basic research

Rheumatoid arthritis(RA)is a common autoimmune disease characterized by progressive joint inflammation and destruction,deformity,loss of mobility,and permanent disability.Although the cellular and molecular mechanisms involved in RA are understood in detail,no drugs or therapies can completely cure RA.Many long-term efforts have been directed towards a better understanding of RA pathogenesis and the development of new classes of therapeutics.Thus,the ongoing elucidation of pathogenic events underlying RA mostly relies on studies of animal models.Herein,we comprehensively review and discuss the characteristics,challenges,and unresolved of issues of various experimental models of RA to provide a basis and reference for the rational selection of experimental RA models for basic investigations into traditional Chinese medicine(TCM).

Anti-angiogenic effect of tripterygium glycosides tablets in animal models of rheumatoid arthritis:A systematic review and meta-analysis

Objectives:To explore and summarize the beneficial effects of a traditional Chinese medicine preparation,Tripterygium glycosides tablets(TGT),in rheumatoid arthritis(RA)animal models of neovascularization,and to provide a reference for future clinical applications and research on its pharmacologic mechanism.Methods:We searched the databases PubMed,Embase,Web of Science,Chinese National Knowledge Infrastructure,VIP,Wan Fang and SinoMed(China Biomedical Document Service System)to identify studies of TGT with outcome indicators of angiogenesis-related factors that were published before April2020.Subgroup analysis and meta-regression were performed for dosage and duration of TGT.Statistical tests and subgroup analysis were conducted using RevMan 5.3,and meta-regression and sensitivity analysis were conducted using STATA/SE 15.0.Results:Fourteen studies of TGT in RA rats were included in this analysis.Treatment with TGT significantly reduces synovial microvessel density and the expression of vascular endothelial growth factor(VEGF),VEGF receptor 2,hypoxia inducible factor a,c-Fos,c-Jun,angiopoietin-1 and angiopoietin-2 compared with control groups(P<.05).Subgroup analysis did not show a significant association of the mRNA levels of VEGF in synovium,assessed using quantitative real-time PCR,with duration or dosage of TGT.Meta-regression analysis also indicated that the effects of dosage and duration were not significantly associated with differences in VEGF mRNA levels.Sensitivity analysis on VEGF m RNA levels did not fundamentally change the results.Conclusions:TGT can reduce synovial neovascularization by decreasing synovial microvessel density and expression of VEGF,VEGF receptor 2,hypoxia-inducible factor a,c-Fos,c-Jun,Ang-1 and Ang-2,thereby suppressing pannus formation and bone destruction in rat models of RA.Additional well-designed studies are required to confirm these findings.

Attenuation of Collagen Induced Arthritis by Centella asiatica Methanol Fraction via Modulation of Cytokines and Oxidative Stress

Objective To investigate the anti-inflammatory, antioxidant and anti-arthritic effects of Centello asiatica methanolfraction （CAME） on collagen-induced arthritis （ClA）, an animal model of rheumatoid arthritis. Methods Arthritis was induced in female wistar rats by immunization with porcine type II collagen. The CIA rats were treated orally with CaME （50, 150, and 250 mg/kg/day） for 15 d （beginning on day 21 of the experimental period）. The clinical, histological, biochemical, and immunological parameters were assessed. Results CaME treatment （150 and 250 mg/kg） significantly attenuated the severity of CIA and reduced the synovial inflammation, cartilage erosion, and bone erosion as evident from both histological and radiographic data. The escalated plasma levels of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-12 alongwith nitric oxide in CIA rats decreased significantly on CaME treatment. The serum levels of type-Ⅱ collagen antibody were significantly lower in rats of CaME （150 and 250 mg/kg） treated group than those in the arthritic group. Furthermore, by inhibiting the above mediators, CaME also contributed towards the reversal of the disturbed antioxidant levels and peroxidative damage. Conclusion Our results clearly indicate that oral administration of CaME suppresses joint inflammation, cytokine expression as well as antioxidant imbalance, thereby contributing to an amelioration of arthritis severity in CIA rats.

Huangqi Guizhi Wuwu Decoction suppresses inflammation and bone destruction in collagen-induced arthritis mice

Objective: Rheumatoid arthritis(RA) is a chronic inflammatory and destructive arthritis, characterized by inflammatory infiltration and bone destruction. Huangqi Guizhi Wuwu Decoction(HGWD) is traditional Chinese medicine, which has been applied in the treatment of RA in clinical. The aim of this study was to investigate the therapeutic effect of HGWD on collagen-induced arthritis(CIA) mouse model.Methods: DBA/1J female mice were used to establish the collagen-induced arthritis(CIA) model. HGWD was administered intragastrically once a day for four weeks starting on the 22nd day after the first immunization. The body weight, hind paw thickness and clinical score were measured every five days. Gait analysis, histopathological staining, enzyme-linked immunosorbent assay(ELISA), ultrasound imaging and micro-computed tomography imaging were performed to determine the effects of HGWD treatment on inflammation and bone structure in this model. Moreover, Real-time PCR and Western blot analysis were used to detect inflammatory factors m RNA and protein levels after HGWD intervention in RAW264.7 cells.Results: HGWD attenuated symptoms of arthritis, suppressed inflammatory synovium area and the serum levels of inflammatory factors, inhibited joint space enlargement in the knee and ankle joints,reduced numbers of osteoclasts, protected bone destruction, as well as improved motor function.HGWD decreased the expression of m RNA for inflammatory factors and the protein expression levels of p-NF-ΚB and IL-17.Conclusion: These results suggested that HGWD suppresses inflammation, attenuates bone erosion and maintains motor function in collagen-induced arthritis mice.

What qualifies as rheumatoid arthritis?

Expansion of diagnostic criteria for rheumatoid arthritis and deletion of exceptions increases sensitivity, but at the expense of specificity.Two decades later, modification of criteria included the caveat: "absence of an alternative diagnosis that better explains the synovitis."That puts great faith in the diagnostic skills of the evaluating individual and their perspectives of disease.The major confounding factor appears to be spondyloarthropathy, which shares some characteristics with rheumatoid arthritis.Recognition of the latter on the basis of marginally distributed and symmetrical polyarticular erosions, in absence of axial(odontoid disease excepted) involvement requires modification to avoid failure to recognize a different disease, spondyloarthropathy.Skeletal distribution, pure expression of disease in natural animal models and biomechanical studies clearly rule out peripheral joint fusion(at least in the absence of corticosteroid therapy) as a manifestation of rheumatoid arthritis.Further, such studies identity predominant wrist and ankle involvement as characteristic of a different disease, spondyloarthropathy.It is important to separate the two diagnostic groups for epidemiologic study and for clinical diagnosis.They certainly differ in their pathophysiology.

A comprehensive meta-analysis of the association between three <i>IL</i>1B polymorphisms and rheumatoid arthritis

Rheumatoid arthritis (RA) is an immune-mediated chronic inflammatory disease that causes huge destruction to human body. IL1B encodes key mediator IL-1β protein, which plays an important role in the pathogenesis of inflammatory syndromes. The aim of this study was to evaluate the association between IL1B polymorphisms and RA. A meta-analysis was performed on the association between three IL1B polymorphisms (IL1B-31: rs1143627;IL1B-511: rs16944;IL1B + 3954: rs1143634) and RA. A trend of significant association was observed between IL1B + 3954 and RA (p = 0.06, odd ratio (OR) = 1.19, 95% confidential interval (CI) = 1.00-1.42). A significant association was found in Europeans under the dominant model between IL1B-511T and RA (p = 0.03, OR = 0.89, 95% CI = 0.81-0.99). Our meta-analysis indicated that IL1B ? 511-T played a protective role against RA in Europeans, and that IL1B + 3954-T had the potential to increase the risk of RA. Future large-scale studies should be considered to confirm the association between IL1B polymorphisms and RA.

Nuciferine alleviates collagen-induced arthritic in rats by inhibiting the proliferation and invasion of human arthritis-derived fibroblast-like synoviocytes and rectifying Th17/Treg imbalance

Rheumatoid arthritis(RA)is a chronic autoimmune disorder marked by persistent synovial inflammation and joint degradation,posing challenges in the development of effective treatments.Nuciferine,an alkaloid found in lotus leaf,has shown promising anti-inflammatory and anti-tumor effects,yet its efficacy in RA treatment remains unexplored.This study investigated the antiproliferative effects of nuciferine on the MH7A cell line,a human RA-derived fibroblast-like synoviocyte,revealing its ability to inhibit cell proliferation,promote apoptosis,induce apoptosis,and cause G1/S phase arrest.Additionally,nuciferine significantly reduced the migration and invasion capabilities of MH7A cells.The therapeutic potential of nuciferine was further evaluated in a collagen-induced arthritis(CIA)rat model,where it markedly alleviated joint swelling,synovial hyperplasia,cartilage injury,and inflammatory infiltration.Nuciferine also improved collagen-induced bone erosion,decreased pro-inflammatory cytokines and serum immunoglobulins(IgG,IgG1,IgG2a),and restored the balance between T helper(Th)17 and regulatory T cells in the spleen of CIA rats.These results indicate that nuciferine may offer therapeutic advantages for RA by decreasing the proliferation and invasiveness of FLS cells and correcting the Th17/Treg cell imbalance in CIA rats.

Polyherbal formula SC-E3 inhibits rheumatoid arthritis activity in a mouse model of type-Ⅱcollagen-induced arthritis

Objective:SC-E3 is a polyherbal formula that contains five medicinal herbs used frequently in traditional herbal medicine.In our previous study,we demonstrated the antioxidant and anti-inflammatory effects of SC-E3.The present study examined the effects of SC-E3 in a mouse model of type-II collagen-induced arthritis(CIA).Methods:In vivo,male DBA/1 J mice were immunized by intradermal injection of bovine type-II collagen and complete or incomplete Freund’s adjuvant,to induce arthritis.SC-E3 was orally administered daily for 23 days.In vitro,bone marrow-derived macrophages(BMMs)were treated with macrophage colony-stimulating factor(M-CSF)and receptor activator of nuclear factor-j B ligand(RANKL)in the absence or presence of SC-E3.Results:Administrations of SC-E3 were found to have anti-arthritic effects in the joints of CIA mice,as evidenced by reduced paw swelling,bone erosion and deformation,inflammatory cell infiltration,and inflammation in synovial membrane.SC-E3 also reduced serum levels of tumor necrosis factor-a,interleukin-1 b,aspartate aminotransferase and alanine aminotransferase.Furthermore,tartrateresistant acid phosphatase-positive osteoclast numbers in the joints were significantly lower in SC-E3-treated CIA mice than in CIA mice.In addition,the differentiations of BMMs to multinucleated osteoclasts induced by M-CSF and RANKL stimulation were dose-dependently reduced by SC-E3.Conclusion:These results suggest that SC-E3 possesses substantial anti-arthritic activity because it inhibits pro-inflammatory cytokines and osteoclastogenesis,and that SC-E3 has potential therapeutic use for the treatment of rheumatoid arthritis.

成纤维细胞生长因子受体1 抑制剂对胶原诱导关节炎模型大鼠骨破坏的影响

背景:课题组前期的研究表明靶向成纤维细胞生长因子受体1(fibroblast growth factor receptor 1,FGFR1)可能是治疗类风湿性关节炎的有效靶点。目的:探讨FGFR1抑制剂(PD173074)对胶原诱导关节炎模型大鼠骨破坏的影响。方法:将25只雌性SD大鼠随机分为5组,正常对照组、模型组、甲氨蝶呤组、PD173074低剂量组、PD173074高剂量组。除正常对照组外,其余各组大鼠建立Ⅱ型胶原诱导关节炎模型。造模成功后正常组及模型组大鼠腹腔注射无菌PBS,甲氨蝶呤组药物注射剂量为1.04 mg/kg,PD173074低剂量组和高剂量组药物注射剂量分别为5,20 mg/kg,1次/周。给药4周后取材,观察大鼠临床症状以及关节肿胀情况,踝关节Micro-CT三维重建及分析,观察踝关节病理变化,检测关节周围血管生成情况及核因子κB受体活化因子配体的表达,检测关节滑膜中p-FGFR1、血管内皮生长因子A、抗酒石酸酸性磷酸酶的表达,观察肝、脾、肾病理变化并计算肝、脾、肾指数。结果与结论:①PD173074能够减轻模型大鼠踝关节临床症状及关节肿胀,延缓骨质丢失,改善骨结构,减轻关节滑膜侵袭以及软骨骨侵蚀,降低关节周围破骨细胞数量,抑制关节滑膜组织中的血管生成,降低核因子κB受体活化因子配体的表达,抑制FGFR1磷酸化蛋白、抗酒石酸酸性磷酸酶和血管内皮生长因子A的蛋白表达。②大鼠肝、脾、肾病理观察表明经过PD173074治疗后无明显的毒副作用。③研究证明了FGFR1抑制剂能够延缓Ⅱ型胶原诱导关节炎模型大鼠关节炎症及骨破坏的进展,并抑制血管的生成。初步验证了PD173074在Ⅱ型胶原诱导关节炎模型中的治疗作用,其可能是通过抑制FGFR1磷酸化发挥作用,为寻找类风湿性关节炎新的治疗靶点提供了方向。

加权共表达网络分析与机器学习识别类风湿关节炎滑膜中的关键基因

背景:类风湿关节炎是一种全身的免疫相关性疾病,主要病理特点是关节滑膜炎性增生及关节软骨的破坏,其发病机制目前尚不明确,迫切需要发现新的具有高度敏感性和特异性的诊断标志物。目的:联合使用生物信息学技术及计算机学习算法,识别并筛选类风湿关节炎患者滑膜中的关键基因,构建类风湿关节炎预测模型并进行验证。方法:从基因表达综合数据库中下载3个包含类风湿关节炎患者滑膜的数据集(GSE77298、GSE55235、GSE55457),GSE77298和GSE55235作为训练集,GSE55457作为测试集,共纳入66个样本,其中类风湿关节炎患者滑膜样本39个,正常滑膜样本27个。应用R语言筛选训练集中的差异基因,然后使用加权共表达网络将训练集中的基因模块化,选出关键模块中的特征基因,将差异表达基因和特征基因取交集,交集基因进入下一步机器学习。采用3种机器学习方法:最小绝对值收敛和选择算子算法、支持向量机-递归特征消除和随机森林算法对交集基因进一步分析获得枢纽基因,将枢纽基因再次相交即得到类风湿关节炎滑膜中的关键基因。以关键基因为变量构建预测类风湿关节炎的列线图模型,推测患者发生类风湿关节炎的危险程度,使用受试者工作特征曲线确定类风湿关节炎预测模型及其关键基因的诊断价值。结果与结论:①通过差异分析,训练集中共筛选出差异基因730个,加权共表达网络分析得到特征基因185个,两者交集基因159个;②最小绝对值收敛和选择算子发现枢纽基因4个,支持向量机-递归特征消除发现枢纽基因11个,随机森林发现枢纽基因5个,取交集后获得关键基因2个(TNS3、SDC1);③基于2个关键基因,在训练集及测试集种构建列线图,其校准预测曲线与标准曲线贴合较好,且预测类风湿关节炎发生的临床效能良好;④上述结果证实,基于生物信息及机器学习算法获得的TNS3和SDC1有可能成为类风湿关节炎诊断和治疗的关键靶点。

Pharmacokinetic and pharmacodynamic study of triptolide-loaded liposome hydrogel patch under microneedles on rats with collagen-induced arthritis

Triptolide (TP), a major active component of Triptelygium wilfordii Hook.F. (TWHF), is used to treat rheumatoid arthritis (RA). However, it has a narrow therapeutic window due to its serious toxicities. To increase the therapeutic index, a new triptolide-loaded transdermal delivery system, named triptolide-loaded liposome hydrogel patch (TP-LHP), has been developed. In this paper, we used a micro-needle array to deliver TP-LHP to promote transdermal absorption and evaluated this treatment on the pharmacokinetics and pharmacodynamics of TP-LHP in a rat model of collagen-induced arthritis (CIA). The pharmacokinetic results showed that transdermal delivery of microneedle TP-LHP yielded plasma drug levels which fit a one compartment open model. The relationship equation between plasma concentration and time was C=303.59 x (e(-0.064t)-e(-0.287t)). The results of pharmacodynamic study demonstrated that TP-LHP treatment mitigated the degree of joint swelling and suppressed the expressions of fetal liver kinase-1, fetal liver tyrosine kinase-4 and hypoxia-inducible factor-1a in synovium. Other indicators were also reduced by TP-LHP, including hyperfunction of immune, interleukin-1/3 and interleukin-6 levels in serum. The therapeutic mechanism of TP-LHP might be regulation of the balance between Thl and Th2, as well as inhibition of the expression and biological effects of vascular endothelial growth factor. (C) 2015 Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V.