Protective Effect and Mechanism of n-butanol Extract from Diploclisia glaucescens(B1.)Diels on Rats with Adjuvant Arthritis

[Objectives]To study the protective effect and mechanism of n-butanol extract of Diploclisia glaucescens(B1.)Diels on rats with adjuvant arthritis.[Methods]A rat adjuvant arthritis(AA)model with similarities to a clinical RA(rheumatoid arthritis)patient was used,and the model was made by injection of Complete Freund s adjuvant(CFA).Body mass and joint swelling degree were used as indicators,and the organ index was calculated and the synovial tissue of rats was examined under microscope to evaluate the protective effect of n-butanol extract on arthritis.The effects of n-butanol extract on TNF-α,IL-1βand PGE_(2)contents in rat serum were detected by ELISA kit.[Results]Arthritic rats experienced significant weight loss;the n-butanol extract reduced the joint swelling in rats.It exerted an effect on rat organs and reduced the contents of TNF-α,IL-1βand PGE_(2) in rat serum,and also reduced synovial inflammation in rats.[Conclusions]The n-butanol extract of D.glaucescens can protect rats with adjuvant arthritis by reducing the content of inflammatory factors.

The mechanism of regulating macrophage polarization based on Notch1 signaling pathway to improve joint inflammation in adjuvant arthritis rats

Objective:To study the impact of the Notch1/Jagged1/RBP-Jκ/Hes1 signaling pathway on macrophage polarization and its role in modulating the inflammatory response in rats with adjuvant arthritis(AA).Methods:The rats were randomly divided into three groups(6 rats):the healthy group(NC),the model group(MC),and the Notch1 inhibitor group(FLI).Medication was administered after 12 days of inducing inflammation.After 30 days,the arthritis index(AI)and degree of swelling in the right hind foot joint(E)were measured in each group.The expression levels of CD80^(+)and CD163^(+)cells in peripheral blood macrophages of rats were analyzed by flow cytometry.The standards of IL-4,IL-10,IL-1β,and TNF-α in rat serum were gauged by Enzyme-linked immunosorbent assay.The expression of Notch1,Jagged1,RBP-Jκ,and Hes1 proteins in rat synovial tissue was detected using Western blot.Results:The degree of swelling(E)and arthritis index(AI)in the MC group rats with AA were significantly higher than those in the NC group(P<0.01).CD80^(+)cell expression was significantly higher compared to the control group(P<0.01),while CD163^(+)cell expression was significantly lower than the control group(P<0.01).IL-1βand TNF-α expression levels were significantly elevated(P<0.01),whereas IL-4 and IL-10 expression levels were significantly decreased(P<0.01).Notch1,RBP-Jκ,Jagged1,and Hes1 protein expression levels were significantly increased(P<0.01).In comparison to the MC group,the rats in the Notch1 inhibitor group exhibited a significant reduction in toe swelling and arthritis index(P<0.01).CD80^(+)cell expression was significantly decreased(P<0.01),while CD163+cell expression was significantly increased(P<0.01).IL-1β and TNF-α expression levels were significantly decreased(P<0.05),whereas IL-4 and IL-10 levels were significantly increased(P<0.01).Notch1,Jagged1,Hes1,and RBP-Jκ protein expression levels were significantly decreased(P<0.05).Correlation analysis revealed a positive association between CD80^(+)and Notch1,Jagged1,Hes1,and RBP-Jκ(P<0.01),while CD163^(+)showed a negative correlation with the expression of these proteins(P<0.01).Conclusion:The Notch1/Jagged1/RBP-Jκ/Hes1 signaling axis regulates macrophage polarization to M2 type and reduces inflammation in AA rats.

Effects of injection of anti-corticotropin release hormone serum in the lateral ventricles and electroacupuncture analgesia on pain threshold in rats with adjuvant arthritis

Rat models of adjuvant arthritis were established, and anti-corticotropin release hormone serum injection in the lateral ventricles and electroacupuncture at right Jiaji （EX-B2） were performed. The pain threshold was decreased at 45 and 60 minutes after injection of the anti-corticotropin release hormone serum. Electroacupuncture at Jiaji can resist this effect. Immunohistochemical staining results showed that the expression of corticotropin release hormone in the hypothalamic paraventdcular nucleus was greater in the electroacupuncture ＋ anti-corticotropin release hormone serum group compared with the anti-corticotropin release hormone serum group. The expression of corticotropin release hormone was correlated with the pain threshold. The effect of endogenous corticotropin release hormone in pain modulation can be obstructed by anti-corticotropin release hormone serum. The analgesia of electroacupuncture can partially resist the depressed pain threshold caused by injection of anti-corticotropin release hormone serum. The analgesic effect of electroacupuncture is associated with the corticotropin release hormone content in the hypothalamus.

Experimental Study on Anti-adjuvant Arthritis in Rats of Kadsura coccinea

[Objectives] To investigate the therapeutic effect of Kadsura coccinea on adjuvant arthritis( AA) rats and its mechanism from cytokine levels. [Methods] The complete Freund's adjuvant( CFA)-induced AA model of rats was used. The therapeutic effect of K. coccinea on AA rats was evaluated by the articular swelling,arthritis score,immune organ index and hematoxylin-eosin( HE) staining. The content of inflammatory factors TNF-α,IL-6,and IL-1β in the serum of AA rats was detected by enzyme-linked immunosorbent assay( ELISA) respectively. [Results]Compared with the model group,the total extract of K. coccinea( KCTE) could significantly inhibit the primary and secondary articular swelling of AA rats,lower the arthritis score and immune organ index,and improve the pathological state of ankles of AA rats. In the groups with a high and middle dosage of KCTE( KCTE-H and KCTE-M),KCTE could significantly reduce the content of TNF-α and IL-6 in the serum of rats,exhibiting a certain dose-effect relationship. [Conclusions]KCTE had significant therapeutic effect on the AA of rats,and the mechanism might be associated with the modulation of immune function and the decrease in the secretion of inflammatory cytokines TNF-α and IL-6.

Study on the Analgesic Effect and Mechanism of Zhitong Capsule (止痛胶囊) in Adjuvant Arthritis Rats

Objective: To observe the analgesic effect of Zhitong Capsule (止痛胶囊, ZTC) and study its mechanism in adjuvant arthritis (AA) rats. Methods: Forty-eight male Sprague-Dawley rats were randomly divided into six groups with 8 rats in each group. On the first day, except to those in the normal group that were treated with normal saline, the same amount of Freund's complete adjuvant (FCA) was given through intradermal injection into the right hind paw to all the rats in the other groups. From the 17th day of the modeling on, the rats in groups of ZTC were administered daily through gastrogavage with a dose of 1000, 500, 250 mg/kg respectively, while equal volume of normal saline was given to those in the normal group and model group, and an equal volume of aspirin (ASA) solution was given to rats in the ASA group through gastrogavage for 10 days, once per day, and on the 27th day, the analgesic effect of ZTC was measured with heat withdraw method. The activities and contents of superoxide dismutase (SOD) and lipid peroxides (LPO) in serum were observed by spectrophotometry, and the level of beta-endorphin (β-EP) in hypothalamus were determined by the assay of immunohistochemistry. Results: ZTC showed significant effects on enhancing the pain threshold and at the same time it increased the activities of SOD and reduced the contents of LPO in serum. ZTC could also increase the level of β-EP in hypothalamus. Conclusion: ZTC has analgesic effect and its mechanism is probably related with its effect in inhibiting the level of oxygen free radicals in serum and increasing the level of β-EP of hypothalamus in rats.KEY WORD Zhitong Capsule, adjuvant arthritis, superoxide dismutase, lipid peroxides, β-endorphin

Effect of Electro-acupuncture on Rat Joint Pathomorpnology of Chronic Adjuvant Arthritis Model

Objective:To study the effect of electro-acupuncture (EA) on pathomorphological changes of joints in rat model of chronic adjuvant arthritis. Methods: The rat chronic adjuvant arthritis model was established by subcutaneous injection of 0.1 ml of complete Freund’s adjuvant to the left hind sole. Forty Wistar rats were randomly divided into the model group, the low frequency (2 Hz) EA group, the high frequency EA (100 Hz) group and the body acupuncture group. After being modeled except the model group, the other three groups were treated with EA or body acupuncture in Yanglingquan points (bilateral) for 3 weeks, the left ankle joints and metatarsal joints of rats were taken for pathological examination by fixing with 10% formalin and embedding in paraffin, sectioning and staining with HE. Results: Obvious inflammatory cell infiltration, loosened synovial tissue, damage of articular cartilage and proliferation of synovial cells and granulation tissue were observed in the sections of joints in model rats.

Therapeutic Effect and Mechanism of Caulophyllum robustum Maxim Extract on Adjuvant Arthritis Rats

[Objectives]To study the anti-inflammation and anti-swelling effect of Caulophyllum robustum Maxim( CRM) extract on secondary lesions of adjuvant arthritis( AA),associated inflammatory factors of AA rats and mice immune function. [Methods]Rats were divided into high,middle and low dose CRM extract groups( 69. 23,34. 61,17. 31 mg/kg),methotrexate group( 0. 68 mg/kg),triptolide group( 7. 88 mg/kg),model group and normal control group. The AA rat model was induced by intradermal injection of Freund's complete adjuvant( FCA). After the AA model was established successfully,the paw swelling of AA rats was observed and recorded. The levels of interleukin-1β( IL-1β),interleukin-4( IL-4),interleukin-10( IL-10),Tumor Necrosis Factor-α( TNF-α),interleukin 17( IL-17),interferon( IFN-γ),granulocyte colony-stimulating factor( G-CSF) and transforming growth factor( TGF-β) in the serum of AA rats were detected by enzyme-linked immunosorbent assay( ELISA). The spleen was removed and weighed for detecting the drug influence on the immune organs.Impacts were observed by gavage administration at different dosages of 25 mg/kg,50 mg/kg,100 mg/kg for seven days,adopting carbon granules clearance test,delayed-type hypersensitivity induced by dinitrofluorobenzene( DNFB) coupled with flow cytometry evaluating lymphocytes( CD3^+,CD4^+,CD8^+) activation and serolysin test. [Results] Compared with the model group,the joint swelling,spleen index and inflammatory cytokines( IL-1β,IL-4,IL-10,TNF-α,IL-17,IFN-γ,G-CSF,TGF-β) were reduced significantly( P < 0. 05,P < 0. 01). And the activity of CRM was significantly superior to methotrexate and triptergium. Moreover,the results indicated that different dosages of 25 mg/kg,50 mg/kg,100 mg/kg of CRM could significantly reduce the carbon granules clearance index( K) and phagocytic index( a),inhibit the phagocytosis of mononuclear macrophages,reduce the mice swelling extent and inhibit DTH in mice induced by DNFB,decrease the quantity of serum hemolysin. Percentage of CD3^+,CD4^+,and the ratio of CD4^+,CD8^+ notably decreased,whereas CD8^+ obviously increased. Intragastrica dose of 50 mg/kg,100 mg/kg were acknowledged as more objective inhibiting effect compared with a dosage of 25 mg/kg.[Conclusions] Decreasing inflammatory cytokines content,non-specific immune,humoral immune and cellular immune function may be the therapeutic mechanism for rheumatoid arthritis( RA).

Effect and mechanism of N-acetylcysteine on adjuvant arthritis rats

Objective:To investigate the anti-inflammatory effects of N-Acetylcysteine(NAC)on adjuvant arthritis(AA)rats and the changes of Toll-like receptor 2(TLR2)-inflammatory factors.Methods:50 SD rats,randomly divided into 10 control groups,40 AA model group,after 10 d,which were divided into model group,NAC groups gave low,medium and high dose NAC(lavage respectively 50,100,200 mg/kg)for 16 d,joint swelling degree and arthritis index evaluation NAC therapeutic effect on AA rats;Immunohistochemical staining and Western blot were used to observe the expression of TLR2 protein in synovial tissue of joints.Serum levels of TNF-α,IL-1βand IL-10 were detected by ELISA.Results:Compared with the control group,the joint swelling and arthritis index in the model group were significant,and the joint swelling and arthritis index in the NAC treatment groups were significantly reduced(P<0.05,P<0.01).The expression of TLR2 protein in the synovial tissue of NAC treatment groups was decreased,inflammatory factors TNF-αand IL-1βwere decreased,and IL-10 was increased(P<0.01).Conclusion:NAC can improve the symptoms of arthritis in AA rats,and its mechanism may be related to inhibition of TLR2 and regulation of downstream inflammatory factor pathways in AA rats.

Changes and correlation of inflammatory polarization and Notch pathway in rats with adjuvant arthritis

Objective:To investigate the relationship between inflammatory polarization and Notch pathway in rats with adjuvant arthritis.Methods:Twelve rats were randomly divided into normal(NC)group(n=6)and model(MC)group(n=6).In the model group,complete Freund's adjuvant(0.1ml/rat)was injected into the right hindfoot to induce inflammation.On the 12th day after inflammation,the changes of plantar swelling degree(E)and arthritis index(AI)were observed,and the expressions of inflammatory polarization markers CD68 and CD206 in peripheral blood were detected by flow cytometry.PCR was used to detect the expression of factors related to Notch signal pathway in peripheral blood.Results:Compared with the normal group,the expression of E,AI,CD68,Notch2,Notch3,Notch4 and Delta1 in the model group increased significantly,while the expression of CD206,Notch1,Jagged1 and Jagged2 decreased(P<0.01or P<0.05).The results showed that CD68,toe swelling degree and arthritis index were negatively correlated with Notch1,Jagged1 and Jagged2,CD68,toe swelling degree and arthritis index were positively correlated with Notch2,Notch4 and Delta1,CD206 was positively correlated with Notch1 and Jagged1,Jagged2 and CD206 was negatively correlated with Notch2,Notch4 and Delta1.Conclusion:Notch signal pathway may promote the occurrence and development of AA by regulating inflammatory polarization of macrophages.

Effects of External Use of Pereskia aculeate Miller on Foot Swelling of Adjuvant Arthritis in Rats

[Objectives] To study the anti-inflammatory effects and its possible action mechanism of Pereskia aculeate Miller on rats with adjuvant arthritis( AJ). [Methods] Fifty SD rats( half male and half female) were randomly divided into 5 groups: blank group,model group,positive control group( Glucosidorum Tripterygll Totorum( GTT),12 mg/kg),and P. aculeate high and low dose group( 0. 86 and0. 43 g/m L). Except the blank group,other groups were induced with complete Freund's adjuvant( CFA) to establish the AA rat model. On the 17 th day of modeling,the drug was externally applied for soaking,and the diameter of foot hole and foot joint before and after administration was measured to observe the degree of swelling. The enzyme-linked immunosorbent assay( ELISA) was adopted to determine the inflammatory cytokines interleukin-1β( IL-1β) and tumor necrosis factor-α( TNF-α). [Results] Compared with the model group,the degree of swelling of the foot sole and foot joints was reduced in the P. aculeate high dose group and the positive control group( P < 0. 05). According to ELISA test,compared with the model control group,the serum levels of IL-1β( P < 0. 05) and TNF-α( P < 0. 05) were significantly reduced in the P. aculeate high dose group and the positive control group. Compared with the positive control group,there was no significant difference( P > 0. 05). [Conclusions] P. aculeate has significant inhibitory effects on foot swelling of adjuvant arthritis in rats,and the action mechanism is possibly related to the decrease of IL-1β and TNF-α.

Correlation analysis of changes in miR145‑5p/Smads pathway and macrophage polarization in adjuvant arthritis rats

Objective:To explore the relationship between the changes of miR145‑5p/Smads pathway and macrophage polarization in adjuvant arthritis rats.Methods:Twelve rats were divided into normal group and model group induced by freund's complete adjuvant(0.1 mL/mouse)by random number table method,with 6 rats in each group.The expression of inflammatory polarization markers IL‑8 and CD206 in synovial tissue was detected by enzyme‑linked immunosorbent assay on the 12th day after the formation of arthritis in rats.Western blotting was used to detect the expression of TGF‑β1/Smads pathway factors in synovial tissues.The expression of miR145‑5P,Smads3 and Smads7 in synovial tissue was detected by RT‑qPCR.Results:Compared with normal group,the expression levels of IL‑8,TGF‑β1 and Smad3 in model group were significantly increased(P<0.05);The expression levels of CD206,Smad7 and miR145‑5P were significantly decreased(P<0.01).The correlation results showed that IL‑8 was positively correlated with Smad3(P<0.01),IL‑8 was negatively correlated with Smad7(P<0.05),CD206 was negatively correlated with Smad3(P<0.01)and positively correlated with Smad7(P<0.05).miR145‑5p was negatively correlated with Smad3(P<0.01)and positively correlated with Smad7(P<0.01).Conclusion:miR145‑5p may inhibit the overactivation of TGF‑β1/Smads pathway,regulate macrophage polarization,and inhibit the development of adjuvant arthritis by inhibiting Smad3 expression.

Progresses of studies on acupuncture and pain signals transduction mechanisms in complete Freund's adjuvant arthritis

Advances of studies on the acupuncture and pain signal transduction mechanisms in complete Freud＇s adjuvant arthritis are reviewed from the three aspects, the first messenger of modulating pain signals and the related receptors, the second messenger of modulating pain signals and other factors possibly involved in modulation of pain signal transduction, etc. It is held that modulation of acupuncture for pain signals is a comprehensive course involved in multi-channels, multi-levels, multi-links, and in future, acupuncture analgesic mechanisms for Freud＇s adjuvant arthritis will be more deeply studied by use of more new techniques and new methods.

Improving Effects of Astragalus Polysaccharides on Cardiac Function via Keap1/Nrf2-ARE Signal Pathway in Adjuvant Arthritis Rats

Objective To observe the effect of Astragalus polysaccharides（APS） on cardiac function and expression of Keap1/Nrf2-ARE signal pathway in adjuvant arthritis（AA） rats. Methods Forty-eight rats were randomly divided into normal control（NC） group, model control（MC） group, leflunomide（LEF） group, and APS group. The AA model rats were induced with Freund＇s complete adjuvant, then administration began from day 19 after modeling for 30 d. The paw swelling and arthritic index（AI）, the cardiac function indexes, the expression of Keap1, maf, Nrf2 m RNAs, and HO-1, r-GCS proteins in cardiac tissues were observed. Oxidation-related substances（SOD, MDA, ROS, and TAC） and cytokines（IL-10 and TNF-ɑ） were also determined. Results Compared with NC group, the paw swelling and AI were increased, and the body weight was decreased in MC group. HR, HI, LVSP, LVEDP, and levels of MDA, TAC, ROS, RNS, and TNF-ɑ in MC group were increased, while dp/dtmax and levels of GSH, TRX, and IL-10 were decreased. Compared with MC group, LEF group showed higher HR and ± dp/dtmax, and lower LVEDP. In APS group, the HR, LVSP, and LVEDP were decreased and ±dp/dtmax was increased. TNF-ɑ was decreased, TAC, ROS, MDA, and IL-10 were increased in both LEF and APS groups. APS and LEF groups also showed less MDA, TAC, ROS, and SOD, and the differences were statistically significant. Spearman correlation analysis showed that the cardiac function parameters were positively correlated with Keap1, anti-oxidant indicators, and anti-inflammation cytokines, and negatively correlated with Nrf2, oxidation indictors, pro-inflammation cytokines, and inflammation indictors.Conclusion APS can adjust the expression of Keap1/Nrf2-ARE signal pathway in AA rats, and can also improve their cardiac function. The mechanism may be involved in increasing myocardial anti-oxidant capacity, reducing oxidative stress, and inhibiting inflammation.

Effects of Xinfeng Capsule(新风胶囊) on the Expression of Platelet Derived Growth Factor in Synovium of Adjuvant Arthritis Rats

Objective： To observe the effects of Xinfeng Capsule （新风胶囊, XFC） on platelet parameters in peripheral blood and expression of platelet derived growth factor （PDGF） in synovium of adjuvant arthritis （AA） rats. Methods： A total of 40 male Sprague-Dawley （SD） rats were randomized into 5 groups： normal control （NC）, AA model control （MC）, methotrexate （MTX） treatment, Tripterygium wilfordii polycoride tablet （TPT） treatment, and XFC treatment. Excluding the NC group, the AA model was induced by intracutaneous injection of 0.1 mL Freund＇s complete adjuvant in the right hind limb. Induction of AA and the effects of drug treatments were assessed by voix pedis swelling, arthritis index （AL） body mass, and the pathological changes of joints and cartilage with a light microscopy. Platelet parameters in peripheral blood were detected with an automated hematology analyzer. PDGF in synovium was detected with immunohistochemical methods and PDGF mRNA expression in synovium was detected with reverse transcription polymerase chain reaction. Results： Compared with the NC group, the MC group had significantly increased voix pedis swelling, AI, platelet （PLT） and plateletcrit （PCT） in peripheral blood and PDGF as well as PDGF mRNA in synovium （all P〈0.01） and the joint cartilage was also highly degenerated. Compared with the MC group, the 3 treated groups had significantly decreased voix pedis swelling, AI, PLT, PCT, PDGF, and PDGF mRNA （P〈0.01）. The body mass in the XFC group was significantly higher than those in MTX and TPT groups （P〈0.05）. The levels of PLT, PCT, PDGF, and PDGF mRNA in the XFC group showed a decreasing tendency with no significant difference compared with the M＇IX and TPT groups （P〉0,05）. PDGF and PDGF mRNA of AA rats were positively correlated with voix pedis swelling, AI, PLT, and PCT （P〈0.05 or P〈0.01）. Conclusions： The expression and biosynthesis of PDGF increase in the synovium of AA rats and correlate with voix pedis swelling, AI, PLT, and PCT. XFC can decrease the levels of PDGF, PDGF mRNA, PLT, and PCT, thereby mitigating inflammation induced by platelet activation and reducing voix pedis swelling and the AI in AA rats.

Effects of cryptotanshinone on immune functions in rats with adjuvant arthritis

Background Cryptotanshinone （CT） was originally isolated from the dried roots of Salvia militorrhiza, an herb that is used extensively in Asian medicine and the extracts of this herb have been used in the treatment of several pathologies, including cardiovascular diseases, hematological abnormalities, hepatitis, and hyperlipidemia, but no studies had been carried on the treatment for rheumatic diseases with it. This study aimed to investigate the effects of cryptotanshinone on immune functions in rats with adjuvant arthritis （AA）. Methods Complete Freund＇s adjuvant was used to induce AA in rats. Thymus and spleen was aseptically taken from normal rats and the AA rats. Then a thymus lymphoid cell suspension, splenic lymphoid cell suspension and peritoneal macrophage cell suspension were prepared. After adding CT （0.1 ug/ml, 1.0 ug/ml, 10 ug/ml, 100 ug/ml, 1000 ug/ml） into the suspension, T and B lymphocytes proliferation was determined by 3-（4,5-2 dimethylthiazal-2yl）2,5- diphenyltetrazoliumbromide （MTT） assay. And the activities of interleukin-1 （IL-1） and IL-2 were measured by the mouse lymphocytes proliferation assay. Results Thymic T and splenic B lymphocyte proliferation of the AA rat was significantly lower, and could be stored through using CT in vitro. CT （100ug/ml and 1000ug/ml） increased T or B lymphocytes proliferation in vitro （P 〈0.01）. In AA rats, the levels of IL-1 released by abdominal PMφ significantly increased whereas the level of IL-2 released by T cells decreased in vitro. CT （1000 pg/ml） decreased the production of IL-1 and promoted production of IL-2 in vitro （P 〈0.05）. Conclusions CT can ameliorate the abnormal immunological functions in AA rats.

Effect of Tripterygium Vilfordii on Adrenal Cortex in Rat with Adjuvant Arthritis

This paper reports the effects of Tripterygium Wilfordii (TW) on adrenal cortex in rats with adjuvant artbritis. Forty rats were divided into 5 groups. Adjuvant arthritis (AA) models were made with complete freund's adjuvant (CFA) in groups Ⅰ-Ⅳ. Each of which was treated with sodium carboxyl methyl cellulose, TW, prednisone and cyclophosphamide respecrively.The untreated rats allocated to group,V served as normal. controls. The swelling of AA markedly subsided in groups Ⅱ,Ⅲ and Ⅳ as compasred with group I (P<0.01). whereas no significant differences were noted among groups Ⅱ, Ⅲ and Ⅳ(P>0.05).The obviously increased plasma corticosterone levels and decreased adrenal ascorbic acid levels were observed in group Ⅱ,whereas decreased plasma corticosterone levels and inereased adrenal ascorbic acid levels were noted in group Ⅲ.There was a striking comtrast between groups Ⅱand Ⅲ. The morphological changes of adrenal glands under light microseope revealed hypertrophic adrenal cortices in group Ⅱ,and atrophic adrenal cortices in group Ⅲ.The above findings snggest that that the effect promoting production of corticosterods may be one of mechanism by which TW can effectively treating autoimmune diseases.

Effects and mechanisms of Cryptotanshinone on rats with adjuvant arthritis

Background Cryptotanshinone （CT） is the major active constituent of Salvia miltiorrhiza Bunge. The present study was carried out to investigate the effects of CT on rats with adjuvant arthritis （AA）. Methods AA was induced by the metatarsal footpad injection with complete Freund＇s adjuvant in male Sprague-Dawley rats. The secondary inflammatory reaction was evaluated by hind paw swelling and the polyarthritis index. Activity of interleukin-1 （IL-1） was detected by the concanavalin A-induced thymocytes proliferation assay. The lymphocytes proliferation and IL-2 production were assayed by 3-（4,5-2dimethylthiazal-2yl）2,5-diphenyltetrazoliumbromide （MTT） and activated mouse splenocytes proliferation, respectively. Results Intragastric administration of CT （50 and 100 mg/kg） significantly decreased secondary inflammatory reactions and increased the spleen and thymus index. There was a marked immunologic and inflammatory response in the AA model, which was accompanied by the decrease of thymocyte proliferation and IL-2 production as well as the increase of IL-1 production. CT apparently enhanced thymocyte proliferation and decreased IL-1 production in AA rats. Conclusion These results indicate that CT may exert its anti-inflammatory and immunoregulatory effects through inhibiting lymphocyte proliferation and production of pro-inflammatory mediators.

Effects of cold-damp and hot-damp environment on VEGF and IL-1 expression in joint cartilage cells in adjuvant arthritis in rats

OBJECTIVE:To study the effects of environmental factors on the degree of injury and expression of vascular endothelial growth factor(VEGF) and interleukin-1(IL-1) in cartilage cells of the joint in a rat model of adjuvant arthritis(AA).METHODS:SD rats aged 10 months were randomly divided into 4 groups that varied by temperature and humidity housing conditions and induction of AA:a control group,a model group,a cold-damp group,and a hot-damp group.All groups except the control group were induced with AA.After 4 w,VEGF and IL-1 expression in cartilage cells of ankle joints of hind limbs were observed.RESULTS:Mean area,optical density,and numbers of VEGF-and IL-1-positive cells in the model group,the cold-damp group,and the hot-damp group were significantly higher than that of the control group(all P<0.05).Optical density and positive cell numbers in the cold-damp group and the hot-damp group were significantly higher than that of the model group(all P<0.05).Optical density and positive cell numbers in the hot-damp group were significantly higher than that of the cold-damp group.Bone in the hot-damp and cold-damp groups was severely injured.CONCLUSION:Environmental factors such as high humidity combined with either high or low temperature increase the severity of damage and expression of VEGF and IL-1 in cartilage cells of joints in rats induced with AA.

Effect of Tongbiling (通痹灵) on the Synoviocyte Function in Adjuvant Arthritis Rats

Objective: To study the effect of Tongbiling (TBL) on the proliferation of synovial fibroblast and interleukin1 (IL1), tumor necrosis factor-α (TNF-α) and prostaglandin E 2 (PGE 2) secreted by synoviocytes in adjuvant arthritis (AA) rats. Methods: Synovial fibroblast was derived from culture of tissue piece. The effect of primary synoviocyte culture supernatants on the fibroblast proliferation were assayed and IL1, TNF-α bioactivity and PGE 2 content of supernatants of cultured synoviocytes were measured. Results: TBL could significantly inhibit the synovial fibroblast proliferation (P<0.001), and downregulate IL1, TNFα and PGE 2 productions (P<0.001); indomethacin could obviously promote the synovial fibroblast proliferation(P< 0.001 ). It significantly inhibited PGE 2 production, but further upregulated IL1 and TNFα secreted by synoviocytes (P<0.01). Conclusion: The therapeutical effect of TBL on AA might be associated with its downregulating the secretory function of synoviocyte, then restoring the abnormal proliferation of fibroblast to normal levels.

Anti-arthritis effect of berberine associated with regulating energy metabolism of macrophage through AMPK/HIF-1α pathway

OBJECTIVE To investigate berberine(BBR)attenuates arthritis in adjuvant-induced arthritic(AA)rats associated with regulating the energy metabolism and correcting the polarization of macrophages through activation of AMP-activated protein kinase(AMPK)and inhibition of hypoxia inducible factor 1α(HIF-1α).METHODS AA rats were treated with BBR(40,80,or 160 mg·kg-1)from days 15 to 29 after immunization.The histopathology of ankle joint was examined through hematoxylin-eosin(HE)staining.The concentrations of tumour necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1β,IL-2,IL-17A,interferon-gamma(IFN-γ),monocyte chemotactic protein 1(MCP-1),IL-4,IL-10,transforming growth factor-β1(TGF-β1),ATP,and lactic acid were measured by using ELISA kits.The percentage of M1 and M2 macro⁃phage cells in joint tissues were evaluated by immune-fluorescence.The expressions of p-AMPK and HIF-1αin joint of AA rats were determined according to immunohistochemistry analysis.The migration of macrophage was detected by Transwell assays.The expression of inducible nitric oxide synthase(iNOS),Arginase-1(Arg1),p-AMPK,AMPK and HIF-1αwere examined by Western blotting.The labeled macrophages were observed with laser confocal microscopy.RESULTS BBR relieved signs and symptoms of AA rats and reversed pathological changes.BBR treatment group exhibited decreases in pro-inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-2,IL-17A,IFN-γ,and MCP-1)coupled with increases anti-inflammatory cytokines(IL-4,IL-10,TGF-β1)in the serum.The number of M1 macrophage was reduced,while the number of M2 macrophage was increased in BBR group joint tissues.Moreover,BBR showed marked up-regu⁃lation the expression of p-AMPK and down-regulation the expression of HIF-1αin joint of AA rats.Next in vitro study,we found BBR up-regulated the expression of p-AMPK,Arg1(M2 marker)and down-regulated the expression of HIF-1α,iNOS(M1 marker)induced by LPS in peritoneal macrophages from normal SD rat.Furthermore,BBR treatment inhibited the migration of macrophages stimulated by LPS.The level of ATP was elevated and lactic acid was reduced in LPSinduced macrophages after treated by BBR.However,Compound C significantly attenuated the effects of BBR on acti⁃vated macrophages.CONCLUSION BBR alleviates inflammation by regulating energy metabolism and correcting the polarization of macrophage through AMPK-HIF-1αpathway.BBR might have great therapeutic value for RA.