Anti-inflammatory effects of Solanum procumbens on a low dose complete Freund's adjuvant-induced arthritis rat model

Objective:To investigate the anti-inflammatory and analgesic effects of Solanum procumbens on complete Freund’s adjuvantinduced arthritis rat models.Methods:We isolated and identified five compounds in the ethanolsoluble Solanum procumbens extract(SP)with anti-inflammatory effects,including ursolic acid,β-sitosterol,hexadecanoic acid,cisvaccenic acid,and vanillic acid.Additionally,we investigated the anti-inflammatory effects of SP on rheumatoid arthritis symptoms,including paw volumes,local temperatures,withdrawal latency,and mechanical withdrawal threshold at the hind paw and white blood cell(WBC)number from complete Freund’s adjuvant-induced arthritis rat models.Results:We have successfully established a complete Freund’s adjuvant-induced arthritis rat model at a low dose(1 mg/mL).SP extract significantly reduced paw volumes(P<0.05),prolonged withdrawal latencies(P<0.05),decreased local temperature,and increased the mechanical withdrawal threshold(P<0.05),but only SP extract at the dose of 300 mg/kg significantly decreased WBC numbers.Conclusions:SP extract could be a potential medication candidate with anti-inflammatory effects for arthritis,but it requires further investigation into the mechanism of the SP and its effectiveness on other models as well as clinical trials.

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.

The effects of Fumaderm~ on immunological function and cytokines in a rat model of adjuvant-induced arthritis

Objective To study the therapeutic effect of Fumaderm in Freund’s complete adjuvant-induced arthritis(AIA)in Spraque-Dawley rats.Methods Adjuvant-induced arthritis(AIA)was established by intradermal injection of 0.1 mL of Freund’s complete adjuvant(CFA)in the palmar surface of the right hindpaw and Fumaderm was delivered by oral gavage for 28 days.After CFA injection,the edema of the hindpaw was determined every two days.On 28 days after CFA injection,the lymphocyte subsets of peripheral blood and the cytokines were determined by flow cytometry,meanwhile the histopathological examination of ankle-joints of the animals was performed.Results Fumaderm had a significant therapeutic effect on AIA.The hindpaw swelling was reduced significantly in a dose-dependent manner.The ratio of peripheral blood T lymphocytes was improved obviously.Multiparameter cytokine analysis from peripheral blood CD4+ T cells showed a decrease of proinflammatory cytokines and an increase of anti-inflammatory cytokines in Fumaderm treated animals.A strongly reduced inflammatory response in the joint synovium was observed.Conclusion Fumaderm has potential anti-inflammatory effects on AIA rats.Further investigation is needed to elucidate the molecular mechanism involved in the clinical effect observed in the AIA model.

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.

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.

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.

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.

Human umbilical cord mesenchymal stem cells as treatment of adjuvant rheumatoid arthritis in a rat model

AIM:To investigate the effect of human umbilical cord stem cells,both mesenchymal and hematopoietic(CD34+),in the treatment of arthritis.METHODS:Mesenchymal stem cells(MSCs) and hematopoietic(CD34+) stem cells(HSC) were isolated from human umbilical cord blood obtained from the umbilical cord of healthy pregnant donors undergoing fullterm normal vaginal delivery.MSC,HSC,methotrexate(MTX) and sterile saline were injected intra-articularly into the rat hindpaw with complete freunds adjuvant(CFA) induced arthritis after the onset of disease(day 34),when arthritis had become well established(arthritis score ≥ 2).Arthritic indices were evaluated and the levels of interleukin(IL)-1,tumor necrosis factor(TNF)-α and interferon(IFN)-γ and anti-inflammatory cytokine IL-10 in serum were determined using enzyme-linked immunosorbent assay.Animals of all groups were sacrificed 34 d after beginning treatment,except positive control(PC) which was sacrificed at 10,21 and 34 d for microscopic observation of disease progression.We used hematoxylin,eosin and Masson's trichrome stains for histopathological examination of cartilage and synovium.RESULTS:The mean arthritis scores were similar in all groups at 12 and 34 d post immunization,with no statistical significant difference.Upon the injection of stem cells(hematopoietic and mesenchymal),the overall arthritis signs were significantly improved around 21 d after receiving the injection and totally disappeared at day 34 post treatment in MSC group.Mean hindpaw diameter(mm) in the MSC rats was about half that of the PC and MTX groups(P = 0.007 and P = 0.021,respectively) and 0.6 mm less than the HSC group(P = 0.047),as indicated by paw swelling.Associated with these findings,serum levels of TNF-α,IFN-γ and IL-1 decreased significantly in HSC and MSC groups compared to PC and MTX groups(P < 0.05),while the expression of IL-10 was increased.Histopathological examination with H and E stain revealed that the MTX treated group showed significant reduction of leucocytic infiltrate and hypertrophy of the synovial tissue with moderate obliteration of the joint cavity.Stem cells treated groups(both hematopoietic CD34+ and mesenchymal),showed significant reduction in leucocytic infiltrate and hypertrophy of the synovial tissue with mild obliteration of the joint cavity.With Masson's trichrome,stain sections from the PC group showed evidence of vascular edema of almost all vessels within the synovium in nearly all arthritic rats.Vacuoles were also visible in the outer vessel wall.The vessel became hemorrhagic and finally necrotic.In addition,there was extensive fibrosis completely obliterating the joint cavity.The mean color area percentage of collagen in this group was 0.324 ± 0.096,which was significantly increased when compared to the negative control group.The mean color area percentage of collagen in hematopoietic CD34+ and mesenchymal groups was 0.176 ± 0.0137 and 0.174 ± 0.0197 respectively,which showed a marked decrement compared to the PC group,denoting a mild increase in synovial tissue collagen fibers.CONCLUSION:MSC enhance the efficacy of CFAinduced arthritis treatment,most likely through the modulation of the expression of cytokines and amelioration of pathological changes in joints.

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).

The petroleum ether fraction of Celastrus paniculatus Willd. seeds demonstrates antiarthritic effect in adjuvant-induced arthritis in rats

Objective:Celastrus paniculatus(CP)Willd.is a plant indigenous to India with medicinal properties.It is used in the ayurvedic treatment of inflammatory ailments such as rheumatoid arthritis.The present study was designed to investigate the therapeutic efficacy of the petroleum ether fraction(PCP)obtained from CP seeds on adjuvant-induced arthritis in rats.Methods:Arthritis was induced in SpragueeDawley rats by immunization with Freund’s complete adjuvant(FCA).Arthritis severity was evaluated by arthritis score,paw volume,tibiotarsal joint thickness,body weight,dorsal flexion pain,motility test,stair climbing ability and index of thymus and spleen.Moreover,histopathology of knee joints supported by haematological analysis was used to assess the anti-arthritic action of PCP.The levels of superoxide dismutase(SOD),glutathione(GSH),catalase(CAT),nitric oxide(NO)and malondialdehyde(MDA)in liver were also assessed.Serum samples were collected for estimation of aspartate transaminase(AST),alanine transaminase(ALT)and alkaline phosphatase(ALP).In addition,cytokines such as tumour necrosis factor-alpha(TNF-a)and interleukin-6(IL-6)were estimated in plasma.Results:PCP significantly alleviated arthritic progression with regards to paw swelling,arthritic score,immune organ indices,hyperalgesic effect and body weight.This phenomenon was correlated with significant suppression of overproduction of inflammatory cytokines(TNFa and IL-6),oxidant stress markers(MDA and NO)and cellular enzyme(AST,ALT and ALP)levels versus arthritic rats without treatment.Moreover,PCP restored the decreased levels of SOD,CAT and GSH.Conclusion:Our results suggest that the anti-arthritic properties of PCP may be due to immunosuppressive effects,cytokine regulation,antioxidant effects and bone-protective activities.

Efficacy of static magnetic field for pain of adjuvant arthritis rats

Rats suffering from adjuvant arthritis (AA) were used to examine the effect of a static magnetic field (SMF) upon pain relief. Rats were divided into SMF- treated AA rats, non-SMF treated AA rats and control rats. Following SMF stimulation, we measured blood flow volume in the paw and then reactive speed response to thermal stimulation. The AA groups exhibited significantly lower blood volume and reactivity to thermal stimulation compared to the control group. Compared to non-SMF, SMF exhibited increased blood flow volume in both the tail and paw, along with an increased reactive speed response to thermal stimulation. Our findings suggest that an improved of blood flow and reactive speed response, induced by SMF, appears to be effective for the relief of pain induced by chronic inflammation.

Effects of Alcohol Extract of Giantleaf Ardisia Rhizome on Levels of Inflammatory Factors in Serum of Rats with Adjuvant Arthritis

[Objectives]The purpose of this study was to investigate the effects of ethanol extract of Giantleaf Ardisia Rhizome on the serum levels of interleukin 6(IL-6),tumor necrosis factorα(TNF-α)and interleukin-1β(IL-1β)in adjuvant arthritis(AA)rats induced by Freund’s complete adjuvant(CFA),so as to investigate its role in the treatment of rheumatoid arthritis.[Methods]Rat models of AA were induced by CFA,and they were given with different doses of ethanol extract of Giantleaf Ardisia Rhizome,followed by scoring of systemic and joint inflammation and swelling degree and determination of serum IL-6,TNF-αand IL-1βlevels.[Results]Compared with the model group,the ethanol extract of Giantleaf Ardisia Rhizome significantly reduced the scores of systemic and joint inflammation and swelling degree and the serum levels of IL-6,TNF-αand IL-1βin AA rats(P<0.05,P<0.01).[Conclusions]The ethanol extract of Giantleaf Ardisia Rhizome can suppress the development of AA in rats and the mechanism may be related to the down-regulation of the expression of serum inflammatory factors(IL-6,TNF-α,IL-1β).

Observation on the Therapeutical Effect of Adjuvant Arthritis in Rats with Medicinal Indirect Moxibustion

ＯｂｓｅｒｖａｔｉｏｎｏｎｔｈｅＴｈｅｒａｐｅｕｔｉｃａｌＥｆｆｅｃｔｏｆＡｄｊｕｖａｎｔＡｒｔｈｒｉｔｉｓｉｎＲａｔｓｗｉｔｈＭｅｄｉｃｉｎａｌＩｎｄｉｒｅｃｔＭｏｘｉｂｕｓｔｉｏｎ￥ＺｈｕＢａｉｊｕｎ；ＷａｎｇＹｕａｎｃｈａｏ；ＤａｉＨｕｉｔｉｎ...

Validation of Reference Genes for mRNA Quantification in Adjuvant Arthritis

Real time quantitative PCR (RT-qPCR) requires a method to normalize the expression of target genes against an en- dogenous reference gene. It is known that commonly used housekeeping genes (HKGs) vary tremendously in inflam- matory conditions;however information about the stability and expression of HKGs in chronic inflammatory joint dis- ease such as rheumatoid arthritis (RA) is scarce. The expressional stability of 10 commonly used HKGs was analyzed in the neuronal (spinal cord, dorsal root ganglia) and in the musculoskeletal tissues (tendon, muscle, epiphysis, capsule, periosteum and ankle joint) using RT-qPCR in the rat model of RA. In individual tissues, suitable HKGs were selected by | △Ct| (│Ct control-Ct arthritis│) and further analyzed by using software programs;geNorm and normfinder. We found hypoxanthine-guanine phosphoribosyl tranferase (HPRT) as the most stable gene except ankle joint while glyce-raldehyde-3-phosphate dehydrogenase (GAPDH) was found as the least stable gene in musculoskeletal tissues. In in-flamed ankle joint where no reference gene was found to be stably expressed, an inflammatory cell marker CD3 was used to normalize peptidylprolyl isomerase B (PPIB), the most homogenous HKG identified among the 10 HKGs. The normalized PPIB was then used to analyze the gene expression of neurokinin 1 (NK1), receptor of substance P, a potent pro-inflammatory mediator. We observed a 3.5 fold increase (p = 0.009) in NK1 expression in inflamed ankle joint compared to control. Our results indicate that reference genes stability should be evaluated before using them as refer- ence during inflammatory conditions. In tissues with intense inflammatory cell infiltration, an inflammatory cell marker should be used to normalize the selected reference gene to avoid erroneous results.

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.

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.

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.