DEVELOPMENT AND EVALUATION OF SOLID-LIPID NANO-PARTICLES OF SULFASALAZINE FOR ANTI- RHEUMATIC ACTIVITY

Chronic rheumatoid arthritis (RA) can cause irreversible joint deterioration over time. Solvent-based lipid nanoparticles (SLNs) are widely used as an efficient method to increase the oral bioavailability of poorly soluble medicines like Sulfasalazine. The present study aimed to formulate and evaluation of anti-rheumatic potential of the solid lipid nano-particles (SLNs) of Sulfasalazine. Drug loaded SLNs were formulated and coated with chitosan (CS) for sustained delivery and characterized for particle size, poly dispersity index and in vitro drug release. safety and efficacy profile of optimized batch was analyzed in animal model. Particle size of the optimized formulation was 269±2.45 nm with the PDI of 0.217±0.008 and entrapment efficiency of about 79.9±2.21. The zeta potential of particles was 35.7 mV. Particles had spherical shape with size ranging 100 nm which was determined by TEM analysis. Created formulation showed that the medication was released from the lipid matrix under regulated conditions, with 83.2±1.5% of the drug released in 24 h. Cmax for drug was higher (337±24) when administered as SLNs drug, similarly Tmax was longer when administered as lipid nanoparticles (6Hr), indicating a sustained drug release from SLNs. complete Freund's adjuvant (CFA) activity in rats administered with CS-SSZ-SLN (300mg/kg) equivalent to doses of 300mg/kg SSZ showed reduction in paw edema by day 9 (53.1 ± 1.75% (p<0.005), day 18 (68.68 ± 2.08%) (p<0.001) and 78.24 ± 2.36 % ( p<0.001) on day 21 respectively. Significant increase in the Tmax and the T1/2 values for the nanoparticles, indicates sustained release of the drugs by the SLNs. Sulfasalazine functions by decreasing inflammation, which is likely responsible for lessening the signs and symptoms of inflammatory diseases such rheumatoid arthritis and inflammatory bowel disease.

Mechanical puncture combined with tumor necrosis factor alpha and complete Freund's adjuvant to construct a rat discogenic low back pain model / 中国组织工程研究

BACKGROUND:Intervertebral disc degeneration is an important cause of low back pain.At present,there are many modeling methods for disc degeneration in China and abroad,but there is not a model for low back pain due to disc degeneration. OBJECTIVE:To compare the effect of mechanical puncture combined with tumor necrosis factor α and complete Freund's adjuvant with a conventional disc mechanical puncture alone. METHODS:A total of 18 male adult Sprague-Dawley rats were randomly divided into 3 groups,with 6 animals in each group.No treatment was given in the blank group.Animal models of intervertebral disc degeneration were made in the L4-5 segments of rats in the control using conventional mechanical puncture.In the experimental group,on the basis of mechanical puncture,tumor necrosis factor α+complete Freund's adjuvant was injected into the L4-5 intervertebral discs using a microinjector to establish a model of disc degeneration induced by mechanical puncture combined with inflammatory factors.Four weeks after surgery,the pain threshold of rats was measured by the hot plate method for assessing the perception of heat injury in rats with intervertebral disc degeneration.MRI examination was performed to observe the disc degeneration in each group.ELISA was used to detect the levels of serum tumor necrosis factor α,interleukin 1β,interleukin 6 and prostaglandin E2.Hematoxylin-eosin and Safranin O-fast green staining were used to observe the morphological changes of the disc. RESULTS AND CONCLUSION:In terms of pain,the behavioral pain threshold of the experimental group was continuously decreased,and the levels of serum inflammatory factors were significantly higher compared with the control group.In terms of morphology,the MRI results showed that the L4-5 nucleus pulposus signal completely disappeared in the experimental group.Histopathological results showed that in the control group,the nucleus pulposus was intact,more notochord cells were visible,and some fiber rings were ruptured,while in the experimental group,there are fewer notochord cells and the structure of the nucleus pulposus and fibrous ring is disturbed,with the boundary disappearing.To conclude,mechanical puncture combined with tumor necrosis factor alpha and complete Freund's adjuvant can successfully establish a discogenic low back pain model in rats.This operation is simple and economical to achieve obvious disc degeneration and low back pain,with greatly shortened molding cycle.This model can be used as a reference for studying discogenic low back pain models.

Ameliorating Effect of Capparis aphylla Roth. in Adjuvant-induced Arthritis in rats with Reference to Changes in Reactants RF, CRP, Radiological Erosions

Despite abundant anti-arthritic therapies, there always remains an opportunity to identify novel drugs as well as targets. One such emerging drug Capparis aphylla Roth. acting on inflammatory cascades. This study was planned to investigate the effect of C. aphylla Roth. on inflammatory cascade in rheumatoid arthritis induced in wistar albino rats. The rheumatoid arthritis was induced by injection of 0.2 mL complete Freund’s adjuvant. The animals were randomized into six groups receiving either vehicle (control), injection of 0.2 mL complete Freund’s adjuvant (model) or treatments (indomethacin 100 mg/kg and methanolic extract of C. aphylla Roth. 190, 240, 300 mg/kg) for 21 days. Various hemodynamic parameters, anti-inflammatory parameters, and x-ray and histopathology of synovial joints were carried out. Paw volume, body weight, arthritic index, ESR, RF, CRP, A/G ratio estimated. Treatment with methanolic extract of C. aphylla Roth. Significantly prevented the rise in body weight, serum A/G ratio and increased arthritic index, paw volume, ESR, RF, CRP. The normal architecture of synovial joints was preserved in histopathological analysis by methanolic extract of C. aphylla Roth. treatment. Furthermore, the prevention of damage in joints carried out by methanolic extract of C. aphylla Roth. Altogether, methanolic extract of C. aphylla Roth. prevented architecture and functions of joints by augmenting the inflammation in complete freund’s adjuvant rats. Such promising effects are attributed to inflammation by novel herbal drug C. aphylla Roth.

Research on the mechanisms of ethanol extract of Scutellaria baicalensis Georgi in improving CFA-induced inflammatory pain rats based on 1H NMR metabolomics / 药学学报

The purpose of this study was to investigate the effects of ethanol extract of Scutellaria baicalensis Georgi (SGE) on endogenous metabolites in toes of rats with inflammatory pain induced by complete Freund's adjuvant (CFA) based on 1H NMR metabolomics, which would provide foundation for revealing the effects and mechanisms of SGE in improving inflammatory pain. This animal experiment was approved by the Committee on the Ethics of Animal Experiments of Shanxi University (SXULL2022062). The rats model of inflammatory pain was induced by subcutaneous injection of CFA (0.1 mL), and the effect of low, medium and high doses of SGE (1.5, 3, 6 g·kg-1) on inflammatory pain were explored. The effects of SGE on relieving inflammatory pain was evaluated by mechanical nociceptive thresholds (MNTs) test. Western blot was used to detect the effects of SGE on protein expression of cyclooxygenase-2 (COX-2), nuclear factor kappa-B (NF-κB) and phospho-NF-κB (p-NF-κB). 1H NMR metabolomics was used to analyze the regulatory effects of SGE on endogenous metabolites in the toes of rats with inflammatory pain. The results showed that SGE (6 g·kg-1) could significantly relieve CFA-induced inflammatory pain, and also notably inhibit the protein expression of COX-2, NF-κB and p-NF-κB. SGE could markedly reverse the changes of 8 differential metabolites, such as glycine, glutamine, succinate, phosphorylcholine, etc. The metabolites were involved in eight metabolic pathways, such as glycine, serine and threonine metabolism, alanine, aspartate and glutamate metabolism, glyoxylate and dicarboxylate metabolism, glutathione metabolism, glycerophospholipid metabolism. These results suggest that SGE may relieve inflammatory pain by regulating NF-κB signaling pathway and metabolic abnormality.

Plantar injection of complete Freund's adjuvant induces behavioral and monoamine neurotransmitter changes in comorbid pain and depression model rats / 中国实验动物学报

Objective To observe the effects of plantar injection of complete Freund's adjuvant(CFA)on pain-depressive behavior and changes in hippocampal monoamine neurotransmitters in rats to establish an animal model of related comorbidity.Methods Sixteen male,8-week-old,SPF-grade healthy SD rats were randomly divided into model and control groups with eight rats in each group.In the model group,rats were anesthetized and injected with 100 μL CFA in the left hind paw to induce the comorbid chronic inflammatory pain and depression model.In the control group,rats were injected with the same volume of saline.Pain thresholds were measured using the Von Frey hair and thermal radiation instrument,and depression-like behaviors were assessed using the open field test(OFT),tail suspension test(TST),and forced swim test(FST).Enzyme-linked immunosorbent assays were used to measure 5-hydroxytryptamine(5-HT),dopamine(DA),and norepinephrine(NE)in rat hippocampal tissue.Histological changes in the hippocampal area were observed by hematoxylin-eosin(HE)staining.Results Compared with the control group,the mechanical withdrawal threshold and thermal withdrawal latency in the model group were significantly decreased at 3,7,and 14 days(P<0.01).The total distance in the OFT was significantly reduced at 7 and 14 days(P<0.01),and the time spent in the center zone was significantly decreased at 14 days(P<0.01).Immobility time in the TST was significantly increased at 14 days(P<0.01),and the immobility time in the FST was significantly increased at 7 and 14 days(P<0.05,P<0.01).5-HT,DA,and NE contents in hippocampal tissue of the model group rats were significantly reduced compared with those in the control group(P<0.01),and hippocampal tissue in the model group showed pathological changes,including irregular neuronal shapes,loose and disordered arrangement,increased intercellular space,some unclear cell nuclei,and some neuronal contraction and apoptosis.Conclusions Injection of 100 μL CFA into the footpad causes pain hypersensitivity,depression-like behavior,significant reductions in monoaminergic neurotransmitters in the hippocampus,and histological changes in the hippocampus,effectively simulating the manifestations of comorbid pain and depression,and is an experimental model to study the pathological mechanisms of comorbid chronic inflammatory pain and depression.

Relationship between complete Freund ' s adjuvant-induced chronic pain-induced negative emotions and morphological changes of hippocampal microglia / 解剖学报

Objective To investigate the alteration of mood and hippocampal microglia morphology in a mouse model of chronic inflammatory pain induced by complete Freund' s adjuvant (CFA). Methods Thirty-two male ICR mice were randomly divided into two groups, including normal saline control group (NS) and CFA model group (CFA). The pain model was established by right hindpaw intraplantar CFA injection. The change of mechanical pain threshold after CFA injection was measured by von Frey fiber needle, the locomotor activity and anxiety-like behavior were determined by open field test (OFT), the depression-like behavior was determined by sucrose preference test (SPT) and forced swimming test (FST). The expression of microglia marker ionized calcium binding adaptor molecule-1 (IBA-1) in the hippocampus was determined by immunohistochemistry and its morphological change was analyzed by Sholl analysis. Results Compared with the NS group, the mechanical pain threshold of CFA group decreased significantly (P<0.01). The behavior result showed that the CFA group showed remarkably reduced time in the inner area (P<0.01) compared with the NS group in the open field test; In the sucrose preference test, the percentage of sucrose preference (P<0.01) of CFA mice decreased significantly compared with the NS mice, while the immobility time of CFA mice (P<0.01) increased significantly in the forced swimming test compared with the NS mice. The immunohistochemistry showed that the number of microglia in the dentate gyrus (DG) of CFA mice increased significantly compared with the NS mice. The Sholl analysis result showed that compared with the NS mice, the number of intersections of microglia in hippocampal DG decreased significantly in CFA mice. Conclusion Our finding indicates that the negative emotions in CFA-induced chronic inflammatory pain may be related to the morphological changes of hippocampal microglia in the mice.

A standardized extract of Centella asiatica (ECa 233) prevents temporomandibular joint osteoarthritis by modulating the expression of local inflammatory mediators in mice

Abstract Objectives To investigate the effect of a standardized extract of Centella asiatica (ECa 233), which has anti-inflammatory properties, on the local expression of the transient receptor potential vanilloid 1 (TRPV1), the acid-sensing ion channel subunit 3 (ASIC3), and the calcitonin gene-related peptide (CGRP) in the temporomandibular joint (TMJ) structure 21 days after injecting the TMJ with complete Freund's adjuvant (CFA). Methodology A mouse model was induced by analyzing the CFA-injected TMJ on days 7, 14, and 21. We assessed TMJ histology by the osteoarthritis cartilage grade score. Then, we observed the effect of different ECa 233 concentrations (30, 100, and 300 mg/kg) and of 140 mg/kg ibuprofen doses on TRPV1, ASIC3, and CGRP local expression on day 21. Results Osteoarthritis cartilage scores were 1.17±0.37 and 3.83±0.68 on days 14 and 21, respectively, in the CFA group (n=5). On day 21, TRPV1, ASIC3, and CGRP expression significantly increased in the CFA group. In the ibuprofen-treated group, TRPV1 expression significantly decreased, but ASIC3 and CGRP showed no significant difference. All ECa 233 doses reduced TRPV1 expression, but the 100 mg/kg ECa 233 dose significantly decreased ASIC3 expression. Conclusions TRPV1, ASIC3, and CGRP expression increased in mice with TMJ-OA on day 21. All ECa 233 and ibuprofen doses inhibited pathogenesis by modulating the local expression of TRPV1 and ASIC3. Therefore, ECa 233 was more effective than ibuprofen.

Evolution of mirror-image pain in temporomandibular joint osteoarthritis mouse model

Abstract Mirror-image pain is a kind of pain that occurs on the contralateral side, but its pathogenesis remains unclear. Objective To develop an osteoarthritis mouse model for investigating mirror-image pain through observing nocifensive behaviors, histological changes, and nociceptive activity at days 3, 7, 14, 21, and 28 after the chemical induction of unilateral temporomandibular joint (TMJ) osteoarthritis. Methodology We randomly divided 6-week-old mice into sham and complete Freund adjuvant groups. To induce nocifensive behaviors, we applied 0.04 g of von Frey filament, 10 psi of air puff, and cold acetone on both sides of whisker pads at different days. The histology of TMJ on both sides was observed by hematoxylin/eosin staining and microcomputed tomography scanning. Furthermore, the nociceptive activity was evaluated using the phosphorylated cyclic AMP response element binding protein (pCREB) and a microglia marker at different days in the trigeminal subnucleus caudalis. Results Nocifensive behaviors against mechanical and temperature stimuli on the contralateral side became stronger than the baseline on day 28, in agreement with the elevation of the pCREB and the microglia marker in the trigeminal subnucleus caudalis. Thus, hypernociception on the contralateral side occurred at day 28. Conclusions Clearly, the TMJ model with unilateral osteoarthritis exhibited mirror-image pain. Therefore, this model is useful in investigating the pathogenesis of pain and in developing treatments.

Analgesic Effect of Panlongqi Tablet on Rats with Chronic Inflammatory Pain / 中国实验方剂学杂志

Objective:To observe the analgesic effect of Panlongqi tablet（PLQT） on rats with chronic inflammatory pain， and to explore mechanism of the action preliminarily from the perspective of the nuclear factor kappa-light-chain-enhancer of activated B cells （NF-κB）and mitogen-activated protein kinase（MAPKs） signaling pathways. Method:Rats were induced to establish model of chronic inflammatory pain by complete Freund adjuvant（CFA）， which was divided into normal group， model group， the PLQT 0.16，0.32，0.64 g·kg-1 group， and the ibuprofen 0.05 g·kg-1 group（also positive group）， give the medicine once a day by gavage. Standard Von Frey fiber was used to evaluated the mechanical pain threshold， acetone was used to stimulated rats inflammatory foot to get the cold-induced response score， with the mechanical pain threshold and cold-induced response score to be observed at 1， 2， 3， 4 and 6 h before and after administration on day 1， and at 4 h after administration on day 3-7. The content of PGE2， IL-1， TNF-α in serum， inflammatory foot and 4-5 lumbar spinal cord was detected by enzyme-linked immunosorbent assay（ELISA）. The protein level of MAPKs （p-p38， p-ERK， p-JNK） in lumbar spinal cord 4-5 was detected by Western blot. The expression of NF-κB p65 in the lumbar spinal cord was detected by IFA. Result:Model group had lower mechanical pain threshold and higher cold-induced response score than these in normal group（P<0.01）， while the mechanical pain threshold and cold-induce response score of the model rats were dose-dependent better regulated after administration of PLQT 0.16， 0.32， 0.64 g·kg-1·d-1（P<0.05，P<0.01）， these effect lasted 6 h， of which PLQT groups get the most significant effect on 4 h， however the effect of IBP was similar to that of PLQT medium dose group. In addition， PLQT reduced the abnormal increase of PGE2， IL-1 and TNF-α contents in serum， inflammatory foot and spinal cord of rats in model group， decreased the protein phosphorylation levels of ERK and JNK in spinal cord， and decreased the protein expression of NF-κB p65， that was significant in the PLQT high-dose group（P<0.01）. Conclusion:PLQT had significant analgesic effect on chronic inflammatory pain model rats， which may be related to the inhibition of NF-κB and MAPKs signaling pathways in spinal cord.

Analgesic effect of Fengshi Qutong Capsules on mice with chronic inflammatory pain caused by complete Freund's adjuvant / 中国中药杂志

The aim was to observe the analgesic effect of Fengshi Qutong Capsules(FSQTC) on chronic inflammatory pain in mice, and investigate its effect on p-ERK/COX-2 signal molecular activity. A model of chronic inflammatory pain was induced in mice by complete Freund's adjuvant(CFA). The mice were divided into normal control group, model group, model+FSQTC 0.3, 0.6 and 1.2 g·kg~(-1 )groups, model+positive control drug ibuprofen(IBP, 0.34 mg·kg~(-1)·d~(-1)) group, and normal control+ FSQTC 1.2 g·kg~(-1)group. FSQTC or IBP was given once a day by oral administration. Standard Von Frey fiber was used to evaluate the mechanical pain threshold, and the acetone stimulation was used to induce inflammatory plantar and observe the cold pain reaction scores. The mechanical pain threshold and cold pain reaction scores were observed before administration and 1, 2, 3, 4, 6 h after administration on the first day, as well as 3 h after administration on the 3 rd to 7 th day. The protein levels of PGE_2, COXs-1,2 and p-ERK in the spinal cord of the inflammatory foot and lumbar 4-5 were detected by enzyme-linked immunosorbent assay, Western blot, immunohistochemistry and immunofluorescence. The results showed that the mechanical pain threshold of the model group decreased and the cold pain reaction score increased as compared with the normal group. FSQTC application could dose-dependently increase the mechanical pain threshold and decrease the cold pain reaction score. The effect lasted for 6 h, most significant at 3 h. The effect of ibuprofen was similar to that of the 0.6 g·kg~(-1) dose group. In addition, FSQTC could reduce the abnormally increased protein content of PGE_2, COX-2 and p-ERK in the inflammatory foot and/or spinal cord of the model group, and the effect was most significant in middle and high dose groups. However, it had no effect on COX-1 in the inflammatory foot and spinal cord of mice. The results suggest that FSQTC has ob-vious analgesic effect on chronic inflammatory pain in mice, which may be related to inhibition of p-ERK/COX-2 signaling pathway.

Evaluation of antiarthritic activity of ginkgolic acid against Freund's adjuvant induced arthritic rat model

This study aimed to analyze the antiarthritic activity of ginkgolic acid against the Complete Freund's Adjuvant (CFA)-induced arthritis in rats. Arthritis was induced through an intradermal injection of CFA (0.1 mL) at the right hind footpad of adult Wistar Albino rats. Ginkgolic acid was administered orally at doses of 25 mg/kg and 50 mg/kg, respectively, once daily via gavage for 25 days upon inducing arthritis. Indomethacin was administered orally at a dose of 3 mg/kg twice in a week which served as positive control group. The animals were sacrificed and subjected to biochemical and histopathological analysis upon completion of treatment. Ginkgolic acid was able to reverse the arthritic effect (p < 0.01) induced by CFA in a dose dependent manner. Swelling of paw, thymus and spleen index, serum biomarker levels, and pro-inflammatory cytokines were significantly reduced (p < 0.01) by the acid whereas the antioxidant enzyme activities were remarkably restored. The histopathological findings were in agreement with the biochemical results. The results indicate that the antioxidant and anti-inflammatory properties of ginkgolic acid can be credited to the antiarthritic effects, and it can be promoted as a potential agent for therapeutic use against osteoarthritis

Effect of Xiao Chaihutang on NF-κB Signaling Pathway in Synovial Tissue of CFA Rats / 中国实验方剂学杂志

Objective:To observe the expression of tumor necrosis factor receptor-associated death domain (TARDD), nuclear transcription factor-κB inhibiting protein α(IκBα)IκB kinase-α (IKKα) and nuclear transcription factor (NF)-κB p65 protein in the NF-κB signaling pathway of synovial tissues of complete Freund's adjuvant (CFA) rats after treatment with Xiao Chaihutang (XCHT). Method:In animal experiments, SPF health adult female Wistar rats were used to prepare the CFA animal model of rats with rheumatoid arthritis with Freund's complete adjuvant and cattle Ⅱ collagen type. According to the random number table, the rats were randomly divided into the normal group, the model group, the low-dose XCHT group, the medium-dose XCHT group, the high-dose XCHT group, and the Tripterygium glucosides group. The drugs were given at 7 d after the model was built. Both normal group and model group were given water for injection,and low-dose XCHT group(5.94 g·kg-1),medium-dose XCHT group(11.88 g·kg-1),high-dose XCHT group(23.76 g·kg-1),Tripterygium glucosides group(0.006 3 g·kg-1) were given corresponding drugs by gavage for three times a day, 2 mL/time. The histopathology of rat ankle joint was observed, and the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in the NF-κB signaling pathway in synovial tissue of CFA rats were detected by Western blot. Result:With the increase of the dosage of XCHT, the histopathological score of the right posterior ankle joint of the experimental rats was increased. And in the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in NF-κB signaling pathway in Synovial Tissue of CFA rats, compared with the model group, the statistical results of the low-dose XCHT group showed decreased protein expressions (PPPα, IκB α, NF-κB p65 in the NF-κB signaling pathway were significantly increased (PPα, IκBα, NF-κB p65 key protein expressions in the NF-κB signaling pathway and protein expressions in low-dose XCHT group were obviously lower (PPConclusion:This study shows that as the dose of Xiao Chaihutang increases, it could effectively improve synovitis, and suppress the expressions of key proteins in the inflammatory signaling pathway of NF-κB, thereby preventing inflammation and suppressing bone erosion.

Immunosuppressant effect of Boswellia serrata extract on CFA induced arthritis in rats

Background: Rheumatoid arthritis (RA) is an immune-mediated arthropathy, so for the treatment disease modifying antirheumatoid drugs are required. In this study we are evaluating the immunomodulatory property of Boswellia serrata extract (BSE) as an alternative medicine.Methods: Complete Freund’s adjuvant (CFA), 0.1ml was injected intradermally in the footpad of left hind paw in 36 Wistar rats to induce RA. Animals were divided into 6 groups. BSE in the doses of 45mg/kg, 90mg/kg and 180mg/kg was administered and cyclophosphamide as standard drug. Various parameters as body weight, paw thickness, ankle diameter, paw volume, arthritis index, TNF- ? and histopathological changes were analyzed.Results: Marked reduction in paw thickness, ankle diameter, paw volume, arthritis index and an improved body weight was found in high dose BSE (180mg/kg) group but the effect was lesser than standard drug Cyclophosphamide.Conclusions: BSE has significant potential as an alternative medicine for treatment of autoimmune diseases like rheumatoid arthritis.

Role and mechanism of Epac protein in regulation of inflammatory pain in paraventricular nucleus of hypothalamus / 中国药理学通报

Aim To investigate the role and mecha-nism of exchange protein directly activated by cAMP (Epac) protein in the paraventricular nucleus(PVN) of the hypothalamus in the development of inflammatory pain in rats. Methods Adult SD male rats were cho-sen to establish the model of inflammatory pain through subcutaneous injection of complete Freund's adjuvant(CFA) on the center of left hind foot. Western blot was used to detect the changes of the expression of Ep-ac protein. Thermal withdrawal latency(TWL) was ob-served after the PVN injecting 8p-CPT-2′-O-Me-cAMP (8p-CPT),the agonist of Epac. Then activated down-stream MEK1/2 protein of Epac in PVN was detected using Western blot when the potency was the strongest.Results ① Compared with normal saline(control group),TWL decreased significantly on d 1, d 3, d 5, d 7,d 9 on the ipsilateral foot of CFA group rats(P<0.01),whereas it returned to normal level in d 13;the paw mechanical withdrawal threshold(PMWT) de-creased significantly on d 6,d 8,d 10,d 12 and d 14 (P<0.05);②Compared with the control,the Epac1 protein in CFA group rats began to decrease from d 3, and significantly decreased on d 3 and d 9(P<0.05), however the expression of Epac2 had no significant change, meanwhile p-MEK1/2 protein decreased sig-nificantly on d 3(P<0.05);③Compared with micro-injection of saline into the PVN(Saline group), the heat hyperalgesia of 20 min and 1h decreased signifi-cantly and TWL increased significantly after PVN ad-ministration of 8p-CPT(8p-CPT group)(P <0.05);paraventricular nucleus p-MEK1/2 protein expression increased significantly in 30 min(P <0.05) and re-covered to normal level 2 h after administration. Con-clusion The Epac1-MEK1/2 signaling pathway in the paraventricular nucleus of the hypothalamus may be in-volved in the development of chronic inflammatory pain induced by CFA.

Paeoniflorin-6′-O-benzene sulfonate influences expression of CXCL12/CXCR4 in spleen of adjuvant-induced arthritis / 中国药理学通报

Aim To observe the expression of CXCL12 and its receptor CXCR4 in spleen of rats with adjuvant-induced arthritis (AA) and the effects of paeoniflorin-6′-O-benzene sulfonate (CP-25). Methods AA rats were induced using complete Freund's adjuvant and were randomly divided into normal group,AA group, CP-25 group (50 mg·kg-1) and methotrexate group (MTX,0.5 mg·kg-1),which were treated from d 14 to d 28. HE staining was used to assess the pathologi-cal changes of spleen. The expression of CXCL12 and CXCR4 in spleen was detected by ELISA,immunohis-tochemitry and Western blot. Results CP-25 (50 mg ·kg-1)alleviated the pathological changes of spleen and decreased the expression of CXCL12 and CXCR4 in spleen of AA rats. The pathological changes of spleen and the expression of CXCL12/CXCR4 in spleen revealed a positive correlation. Conclusions Increased expression of CXCL12 and its receptor CX-CR4 may be associated with the pathological changes of spleen in AA rats,which plays an important role in the pathogenesis of RA. CP-25 has obvious therapeutic effect on AA rats and its mechanism may be related to the inhibition of the expression of CXCL12 and CXCR4 in the spleen.

Heliopsis longipes: anti-arthritic activity evaluated in a Freund's adjuvant-induced model in rodents

ABSTRACT This study assesses the anti-arthritic effect of the affinin-enriched (spilanthol, main alkamide) hexane extract from the roots of Heliopsis longipes (A. Gray) S.F. Blake, Asteraceae, on a Freund adjuvant-induced arthritis model in rodents. The extract was orally administered at a dose of 2, 6.6, or 20 mg/kg; a significant edema-inhibitory activity in the acute and chronic phases was observed with a dose of 2 and 20 mg/kg, respectively. The extract showed higher anti-inflammatory and anti-arthritic effects than the reference drug phenylbutazone (80 mg/kg). Moreover, the extract prevented the occurrence of secondary lesions associated to this pharmacological model.

Effects of Different Concentrations of Capsaicine on the Chronic Knee Arthritis Pain Model in Mice / 现代生物医学进展

Objective:To observe the therapeutic effect ofcapsaicin at different concentrations on chronic knee arthritis pain model in mice.Methods:Choosing 50 healthy adult male Kunming mice builded chronic knee arthritis pain model by injecting 0.01 mL CFA (Complete Freund's Adjuvant,CFA) into left joint cavity.The model would be succeed in building after 3 weeks.The successful model mice were divided into five groups randomly (n=10):The first experimental group (saline group),the second experimental group (capsaicin excipient group),the third experimental group (0.5 ％ of capsaicin),the fourth experimental group (3 ％ of capsaicin) and the fifth experimental group (8 ％ of capsaicin).All of the mice would be observed the time of withdrawal latencies from the thermal heated surface after administration of one,four and seven hours,and thermal withdrawal time within 60 days after the injection.Results:①The physiological saline group compared with excipient group,the thermal withdrawal time had no statistically significant difference (P＞0.05)after administration of one,four and seven hours,and thermal withdrawal time within 60 days.②The acute pain duration of the third group would disappear after capsaicin injection 7 hours,four hours for the fourth group,and one hour for the fifth group.③The duration of analgesia of the third group,lasted for 18.9± 1.1 days;The analgesia time of the fourth group lasted for 33.7± 1.0 days;The analgesia time of the fifth group lasted for 58.2± 1.2 days.Conclusions:Capsaicin has analgesic effects on chronic knee pain model in mice induced by CFA,and the days of analgesia increases with the concentration of capsaicin.

Evaluación de la actividad de Baccharis latifolia en modelos de artritis experimental / Evaluation of Baccharis latifolia activity in experimental arthritis models

La artritis reumatoide (AR) es una enfermedad crónica de naturaleza autoinmune e inflamatoria que conduce a la formación de pannus seguido de la destrucción de las articulaciones, se caracteriza por hiperplasia sinovial, inflamación y angiogénesis. La especie vegetal Baccharis latifolia es utilizada tradicionalmente en muchas regiones de nuestro país para tratar el dolor, la inflamación y la artritis. En el presente estudio se evaluó la actividad antiartrítica del extracto etanólico de B. latifolia en modelos murinos de artritis reumatoide inducida por adyuvante, la estimación del edema/espesor de la pata inflamada, parámetros hematológicos (hemoglobina, velocidad de sedimentación globular, recuento de eritrocitos, recuento total de leucocitos) y observación radiológica fueron evaluados. La administración oral del extracto etanólico de B. latifolia (600 mg/kg de p.c.) inhibió significativamente (p<0.001) el incremento del edema/espesor de la pata en el modelo de artritis subcrónica. Del mismo modo, B. latifolia (500 mg/kg de p.c.) inhibió significativamente el incremento del edema/espesor de la pata en el modelo de artritis crónica (p<0.05, p<0.01), los pesos de los animales se mantuvieron sin variación durante el tratamiento. Por otro lado, los parámetros hematológicos señalan que los niveles de hemoglobina disminuyen en ratones artríticos y que esta disminución es revertida tras la administración de los extractos de B. latifolia, este mismo perfil de recuperación es observado tras el recuento de glóbulos rojos. Adicionalmente, la velocidad de sedimentación globular (VSG) incrementada en ratones artríticos, es revertida tras la administración de B. latifolia. El análisis radiológico evidenció el efecto del extracto etanólico de B. latifolia en el retraso de la destrucción ósea. Los resultados sugieren que el extracto etanólico de B. latifolia tiene una potencial actividad antiartrítica.

Rheumatoid arthritis (RA) is a chronic autoimmune and inflammatory disease leading to pannus formation followed by the destruction of the joints; it is characterized by synovial hyperplasia, inflammation and angiogenesis. The plant species Baccharis latifolia is traditionally used in many regions of our country to treat pain, inflammation and arthritis. In the present study the anti-arthritic activity of ethanol extract of B. latifolia was evaluated in murine experimental model of rheumatoid arthritis induced by adjuvant, edema estimation / thickness of the inflamed foot, hematological parameters (hemoglobin, erythrocyte sedimentation rate, erythrocyte count was assessed, total white blood cell) and radiological observation were evaluated. The oral administration of the ethanolic extract of B. latifolia (600 mg / kg p.c.) significantly inhibited (p<0,001) increased edema / paw thickness in the subchronic model of arthritis. Similarly, B. latifolia (500 mg / kg bw) significantly inhibited the increase of edema / paw thickness in the model of chronic arthritis (p <0.05, p <0.01), the weights of the animals were kept without variation during treatment. Moreover hematological parameters indicate that hemoglobin levels decrease in arthritic mice and that this decline is reversed after administration of the extracts of B. latifolia, this same profile is observed recovery after red blood cell count. Additionally, erythrocyte sedimentation rate (ESR) increased in arthritic mice is reversed after administration of B. latifolia. The radiological analysis showed the effect of ethanol extract of B. latifolia in delaying bone destruction. The results suggest that the ethanolic extract of B. latifolia has potential antiarthritic activity.

Potential of Vigna radiata (L.) sprouts in the management of inflammation and arthritis in rats: Possible biochemical alterations.

Vigna radiata (Fabaceae) is an important pulse crop widespread throughout the tropics and warm temperature regions. In this study, we evaluated the in vitro anti-inflammatory and in vivo antiarthritic activity of Vigna radiata sprouts in rats. The in vitro anti-inflammatory activity was determined by membrane stabilization and protein denaturation method. Whereas, the antiarthritic activity of the ethanolic extract of the sprouts was evaluated by complete Freund’s adjuvant model with diclofenac sodium as the standard drug. Body weights, paw volume, biochemical parameters such as lipid peroxidation, total reduced glutathione, myeloperoxidase and lysosomal enzymes like cathepsin-D, N-acetyl β-D-glucosamindase and β-D-glucuronidase were estimated. Treatment with ethanolic extract of V. radiata exhibited significant membrane stabilization activity and protein denaturation activity, and significantly attenuated the biochemical changes induced by administration of complete Freund’s adjuvant. The findings of the present study suggest the possible role of Vigna radiata in the therapeutics of arthritis.

Anti-inflammatory effects of botulinum toxin type A in a rat model of arthritis / 中华物理医学与康复杂志

Objective To verify the anti-inflammatory effects of intra-articular injection of botulinum toxin type A (BoNT/A) on adjuvant-induced arthritis using a rat model.Methods A murine model of chronic ankle arthritis was established in 90 Wistar rats by injection of 0.1 ml of complete Freund adjuvant (CFA) into the pads of their left paws.They were then randomly divided into a BoNT group (n =30) which received an intra-articular injection of 0.1 ml (20 IU) of BoNT/A,an NS group (n=30) which received intra-articular injection of0.1 ml of normal saline solution and a sham group (n =30) which were punctured without any injection.In addition,30 normal rats formed a control group.Infrared thermal imaging was performed and an index of arthritis was evaluated every three days.The infrared thermal imaging revealed the expression of interleukin-1β (IL-1β) through hematoxy-eosin (HE) staining.Results The arthritis index began to increase 3 days after the injection of CFA and it had increased significantly after 10 days,reaching a peak value of 18,24 days after the injection.The infrared thermal imaging showed that the temperature in the right paw increased greatly after the injection.Following the development of arthritis,the temperature declined gradually,arriving at a steady temperature of between 37.5 and 38.0 ℃ in both ankles 20 days after the injection.The average temperature in both paws of the BoNT group had decreased significantly more by 7 and 14 days after the injection than in the NS and sham groups.The expression of IL-1β in the synovium of the ankle joint also had decreased significantly more in the BoNT group after 7 and 14 days.HE scoring showed an obvious histopathologic change in the hypertrophic synovium,inflamnatory cell infiltration,cartilage destruction and exposure of subchondral bone after 7 and 14 days compared with right after the injection in all groups except the control group.Moreover,the average HE scores of the BoNT group rats after 7 and 14 days were significantly lower than those seen in the NS and sham groups at the same time points.Conclusion Intra-articular injection of botulinum toxin type A has an anti-inflammatory effect on arthritis induced by complete Freund adjuvant,at least in rats.