Zanthoxylum bungeanum Aqueous Extract Treated Rheumatoid Arthritis by Inhibiting Synovial Cell Proliferation and Inducing Cell Apoptosis / 中国实验方剂学杂志

ObjectiveTo investigate the therapeutic effect of the water extract of Zanthoxylum bungeanum aqueous extract（ZBAE）on rheumatoid arthritis. MethodThe sixty SD rats were divided into normal group， model group ［complete Freund's adjuvant （CFA）， 10 mg·kg-1］， methotrexate（MTX） group （0.25 mg·kg-1）， low -， medium -， and high-dose ZBAE groups （90， 180， 360 mg·kg-1）. The rats in MTX group were given intraperitoneal injection for two weeks， three times a week， and the rats in ZBAE group were administrated for 14 days. The swelling of the ankle joint and body weight were observed， and arthritis scores were also performed. Computed tomography （CT）， hematoxylin-eosin （HE） staining and safranine-O and fast green staining were used to observe the effect of ZBAE on synovial hyperplasia and bone protection. Cell counting kit-8（CCK-8）method was used to detect the proliferation of the RA-FLSs cells treated with ZBAE. According to the results of CCK-8 experiment， the optimal concentration and time of administration were determined， blank group， low -， medium -， and high-dose ZBAE groups （0.08，0.10，0.12 g·L-1） were set up. The cell cycle distribution and apoptosis rate were analyzed by flow cytometry，the migration ability of RA-FLSs cells was examined by scratch test. Western blot was used to detect the effect of ZBAE on phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， cyclin-dependent kinase 2 （CDK2）， Cyclin A and phosphorylated PI3K， Akt （p-PI3K，p-Akt） protein expression in RA-FLSs cells. ResultCompared with the normal group，joint swelling index and arthritis score were increased in the model group （P<0.05），the bone of the ankle was seriously damaged， and there was obvious synovial hyperplasia. Compared with the model group， the ZBAE group could significantly reduce the joint swelling index （P<0.05）， inhibit synovial hyperplasia and cartilage destruction. In vitro study showed that compared with the blank group， ZBAE could inhibit the migration of RA-FLSs （P<0.05）， promoted cell apoptosis （P<0.05）， and acted on RA-FLSs cells in S phase to inhibit cell proliferation. Moreover， the result of Western blot showed that compared with the blank group， the expression of p-PI3K， p-Akt， CDK2 and Cyclin A proteins were significantly decreased in the high dose group of ZBAE （P<0.05）. ConclusionThese results suggest that ZBAE has a therapeutic effect on rheumatoid arthritis by inhibiting synovial hyperplasia， promoting synovial apoptosis and inhibiting its migration.

Paeoniflorin-6'-O-benzene sulfonate ameliorates progression of adjuvant-induced arthritis by inhibiting interaction between Ahr and GRK2 of fibroblast-like synoviocytes / 中国药理学与毒理学杂志

OBJECTIVE Aryl hydrocarbon receptor (Ahr) is thought to be a crucial factor that regulates immune responses, which may be involved in the pathogenesis of autoimmune inflammation including rheumatoid arthritis (RA). The results of our group in recent years have shown that CP-25, a novel ester derivative of paeoniflorin, has a good effect on improving RA animal models. However, whether the anti-arthritis effect of CP-25 is related to Ahr remains unclear. METHODS CP-25 treatment ameliorated adjuvant-induced arthritis (AA), a mouse model of RA, by inhibiting Ahr-related activities in fibroblasts like synoviocytes (FLS). AA rats were treated with CP-25 or paroxetine from day 17 to 33 after immunization. RESULTS CP-25 alleviated arthritis symptoms and the pathological changes, decreased the expression of Ahr in the synovium and FLS of AA rats. Besides, treatment with CP-25 reduced the proliferation and migration of MH7A caused by Ahr activation. In addition, we also demonstrated that CP-25 down-regulated the co-expres?sion and co-localization of Ahr and G protein-coupled receptor kinase 2 (GRK2) in MH7A. CONCLUSION The data pre?sented here demonstrated that CP-25 suppressed FLS dysfunction in rats with AA, which were associated with reduced Ahr activation and the interaction between Ahr and GRK2.

Study on Effects and Active Ingredients of Tibetan Medicine Lamiophlomis rotata against Rheumatoid Arthritis / 中国药房

OBJECTIVE：To study the effects and active ingr edients of Tibetan medicine Lamiophlomis rotata against rheumatoid arthritis （RA）. METHODS ：Fifty-six SD rats were randomly divided into normal control group （0.5％ sodium carboxymethylcellulose），model group （0.5％ sodium carboxymethylcellulose ），methotrexate group （positive control group ，3 mg/kg），L. rotata water extract low-dose ，medium-dose and high-dose groups （0.5，1，2 g/kg，by crude drug ），L. rotata total flavonoid group （200 mg/kg，by flavonoid extat ），with 8 rats in each group. Except for normal control group ，other groups were given Freund ’s complete adjuvant （FCA）on the rat ’s right hind footpad to induce adjuvant-induced arthritis model. The next day after injection of FCA ，rats in all groups were given relevant medicine intragastrically ，once a day （once every 3 days for methotrexate group ）for 30 days. At 15th and 30th day of administration ，the degree of paw swelling of left hind foot was measured，and the arthritis index ，spleen index were calculated. At the end of 30th day of administration ，the levels of TNF-α，IL-1β， IL-6 and IL- 10 in rat serum were determined by ELISA assay ，the thymus index and spleen index were calculated ，the pathological changes of the ankle joints were observed by HE staining. RESULTS ：Compared with normal control group ，degree of paw swelling，arthritis index at 15th and 30th day of administration as well as the spleen index and the levels of TNF-α，IL-1β， msxmX0146） IL-6 in serum at 30th day of administration were significantly com increased in model group （P＜0.01）；while the thymus indexyunbinji- and IL- 10 level at 30th day of administration were ang@swu.edu.cn significantly decreased（P＜0.01）；synovial cell proliferation and infiltration of articular cavity were observed in ankle joint. Compared with model group ，degree of paw swelling ，arthritis index at 15th and 30th day of administration as well as the spleen index and the levels of TNF-α，IL-1β，IL-6 in serum at 30th day of administration were significantly decreased in each medicine group （P＜0.01）；while the thymus index and IL- 10 level at 30th day of administration were significantly increased （P＜0.01）；the pathological changes of arthritis were significantly improved. Compared with L. rotata water extract high-dose group ，there were no significant differences in degree of paw swelling at 15th day of administration as well as the arthritis index ，spleen index ，levels of inflammatory cytokines and pathological changes of ankle joint in L. rotata total flavonoid group （P＞0.05）. CONCLUSIONS ：Tibetan medicine L. rotata shows well anti-RA activity ，and total flavonoids may be the active ingredients of its efficacy.

Anti-inflammatory Effect of Gentiopicroside in Adjuvant-induced Arthritis Rats / 中国实验方剂学杂志

Objective::To study the therapeutic and inflammatory effects of gentiopicroside(GPS) on adjuvant-induced arthritis (AA) rats. Method::The 36 male SD rats were randomly divided into six groups, namely the model group, GPS groups (low, medium and high dose), and the methotrexate (MTX) group, with six rats in each group. AA rats were induced through intradermal injection with 0.1 mL complete Freund's adjuvant (CFA) into the right hind paw, except the normal group. After modeling, rats in each group were treated with drugs for 7 days, once a day. The doses were 30, 60, 120 mg·kg-1 in the GPS groups, and 0.2 mg·kg-1 in the MTX group. The normal group and the model group were intragastrically treated with the same volume of normal saline. During the experiment, the paw thickness and paw volume of rats were recorded everyday by the digital vernier calipers and the toe volume measuring instrument. On the seventh day, X-ray and histopathological examination of the ankle joints were performed by the small animal living imaging instrument and hematoxylin eosin stain. Blood samples were collected from the abdominal aorta at the end of the experiment to determine the levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α in serum by enzyme-linked immunosorbent assay(ELISA) kits. The mRNA levels of IL-6 and TNF-α in synovial tissues were determined by Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR). Result::Compared with the normal group, the results of each index in the model group were significantly different (P<0.01). Compared with the model group, the results of paw volume and paw thickness decreased significantly (P<0.01), TNF-α level decreased significantly (P<0.01), and IL-6 and TNF-α mRNA levels decreased significantly (P<0.05, P<0.01) in drug treated groups. The results of X-ray and histopathological examinations indicated that GPS had a protective effect on the ankle joints of AA rats. Conclusion::GPS has the therapeutic effect on AA rats by inhibiting levels of proinflammatory cytokines in serum and relevant mRNA levels in synovial tissues.

Deciphering the pharmacological mechanism of Guan-Jie-Kang in treating rat adjuvant-induced arthritis using omics analysis / 医学前沿

Traditional Chinese medicine (TCM) formulas have attracted increasing attention worldwide in the past few years for treating complex disease including rheumatoid arthritis. However, their mechanisms are complex and remain unclear. Guan-Jie-Kang (GJK), a prescription modified from "Wu Tou Decoction," was found to significantly relieve arthritis symptoms in rats with adjuvant-induced arthritis after 30-day treatment, especially in the 24 g/kg/day group. By analyzing 1749 targets related to 358 compounds in the five herbs of GJK, we identified the possible anti-arthritis pathways of GJK, including the calcium signaling and metabolic pathways. Bone damage levels were assessed by micro-computed tomography, and greater bone protective effect was observed with GJK treatment than with methotrexate. Receptor activator of nuclear factor κB ligand (RANKL)-RANK signaling, which is related to calcium signaling, was significantly regulated by GJK. Moreover, a target metabolomics assay of serum was conducted; 17 metabolic biomarkers showed significant correlations with treatment. An integrated pathway analysis revealed that pyruvate metabolism, purine metabolism, and glycolysis metabolism were significantly associated with the effects of GJK in arthritis treatment. Thus, this study establishes a new omics analytical method integrated with bioinformatics analysis for elucidating the multi-pathway mechanisms of TCM.

Study on Toxicity Mechanism of Aconitum carmichaeli Lipid-soluble Alkaloids to Adjuvant-induced Arthritis Model Rats Based on Plasma Metabolomics / 中国药房

OBJECTIVE：To study the toxicity mechanism of lipid-soluble alkaloids of Aconitum carmichaeli to adjuvant-induced arthritis （AIA） model rats. METHODS： Totally 40 rats were randomly divided into blank group （ultrapure water）， model group （ultrapure water） and A. carmichaeli lipid-soluble alkaloids low-dose and high-dose groups （12.5， 35 mg/kg）， with 10 rats in each group. Except for blank group， rats in other groups were given complete Freund’s adjuvant 0.1 mL on the right hind paw to induce AIA model. 19 d after modeling， they were given relevant medicine intragastrically， once a day. After 14 d of administration， endogenous metabolites were separated and identified from plasma by UPLC-LTQ/Orbitrap MS. Then， the collected data were analyzed by principal component analysis （PCA） and partial least squares-discriminant analysis （PLS-DA）. Variable importance projection （VIP）＞1 and P value （＜0.05） were used to screen differential metabolites in plasma. Retrieving the database of Kyoto Encyclopedia of Genes and Genomes according to the differential metabolites，the toxic mechanism of A. carmichaeli liposoluble alkaloids to AIA rats were speculated. RESULTS： A total of 57 plasma metabolites were indentified， and 11 differential metabolites such as L-proline， 6-hydroxynicotinic acid and adenosine were identified. After inducing AIA model， the plasma contents of L-proline and uridylic acid were decreased significantly （P＜0.05 or P＜0.01）， and the content of deoxycytidine was increased significantly （P＜0.01）. Low dose of A. carmichaeli lipid-soluble alkaloids could decrease the plasma contents of adenosine and L-proline in rats （P＜0.05 or P＜0.01）， while the plasma contents of deoxycholic acid was increased significantly （P＜0.05）. High dose of A. carmichaeli lipid-soluble alkaloids could decrease the plasma contents of 6-hydroxynicotinic acid， adenosine， carnitine， L-proline， N-formylaminobenzoic acid were decreased significantly （P＜0.05 or P＜0.01）， while the plasma contents of deoxycholic acid， L-arginine， deoxycytidine and L-lysine were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： The toxicity of low-dose of A. carmichaeli lipid-soluble alkaloids to AIA model rats is less； the toxicity of high-dose of A. carmichaeli lipid-soluble alkaloids to AIA model rats may be related to abnormal bile secretion， lysine biosynthesis and metabolic disorders of purine， pyrimidine， tryptophan， proline and arginine.

Effect and Mechanism of Dihydroartemisinin on Rheumatoid Arthritis Animal Models / 中国实验方剂学杂志

Objective:To study the protective effect of dihydroartemisinin (DHA) on adjuvant-induced arthritis (AIA) and collagen-induced arthritis (CIA) in rats, in order to explore its possible mechanism. Method:Wistar rats were randomly divided into eight groups, namely AIA control group, AIA model group, AIA DHA group and AIA methotrexate group, CIA control group, CIA model group, CIA DHA group and CIA methotrexate group. To establish adjuvant-induced arthritis (AIA) model, rheumatoid arthritis rats were induced through intradermal injection with 0.1 mL Freund's complete adjuvant (FCA) into right postpedes except for the control group. To establish the model of collagen-induced arthritis (CIA), except for control group, the caudal root of rats was immunized subcutaneously with 0.2 mL of emulsion containing 1 g·L-1 of Collagen type Ⅱ (CⅡ). One week later, CⅡ emulsion was injected for the second time. After the rheumatoid arthritis model was successfully established and the administration with DHA (30 mg·kg-1·d-1), the anti-inflammatory effect of DHA on AIA/CIA rats was observed, including the arthritis index (AI), paw swelling degree and effect of DHA on immune organ index of AIA/CIA rats. Interleukin (IL)-6 levels in serum was detected by enzyme-linked immunosorbent assay (ELISA) and pathological sections of ankle joints of AIA/CIA rats. RAW264.7 macrophage cells were cultured in vitro and treated with DHA at various doses (0.5, 1, 2, 4, 8, 16 μmol·L-1) for 24 h, and the cell viability was detected by methyl thiazolyl tetrazolium(MTT) assay. Lipopolysaccharides (LPS) group, LPS+DHA groups (0.5, 1, 2 μmol·L-1) and control group were established. The level of IL-6 was detected by enzyme-linked immunosorbent assay(ELISA). The protein expression levels of nuclear transcription factor-κB p65 (NF-κB p65) was tested by Western blot. Result:Compared with control group, the paw swelling, AI, spleen index and IL-6 levels of model group were significantly increased (PPPPPPPP-1) groups had a remarkable effect on the cell viability (PP-1. The level of IL-6 and the protein expression of NF-κB p65 in LPS group were higher than those of control group. Compared with LPS group, DHA (0.5 μmol·L-1) groups could significantly reduce the secretion of IL-6 (PκB p65. Conclusion:DHA can alleviate the ankle joint lesion on rheumatoid arthritis rats. Its mechanism may be related to NF-κB signal pathway.

Anti-arthritic Effects of Oplopanax elatus in a Rat Model of Rheumatoid Arthritis (Adjuvant-induced Arthritis)

The stems of Oplopanax elatus (OE) have long been used to treat inflammatory disorders in herbal medicine, and in the previous investigation, OE was found to possess anti-inflammatory activity in lipopolysaccharide-treated macrophages, RAW 264.7 cell. OE reduces inducible nitric oxide (NO) synthase-induced NO production, and interferes with mitogen-activated protein kinase activation pathways. In the present study, the pharmacological action of the water extract of OE was examined to establish anti-arthritic action, using a rat model of adjuvant-induced arthritis (AIA). The water extract of OE administered orally inhibited AIA-induced arthritis at (100 – 300) mg/kg/day. The paw edema was significantly decreased, in combination with reduced production of pro-inflammatory cytokines. The action mechanism includes an inhibition of MAPKs/nuclear transcription factor-κB activation. These new findings strongly suggest that OE possesses anti-arthritic action, and may be used as a therapeutic agent in inflammation-related disorders, particularly in arthritic condition.

Madecassoside impedes invasion of rheumatoid fibroblast-like synoviocyte from adjuvant arthritis rats via inhibition of NF-κB-mediated matrix metalloproteinase-13 expression / 中国天然药物

Fibroblast-like synoviocytes (FLS) play a pivotal role in Rheumatoid arthritis (RA) pathogenesis through aggressive migration and invasion. Madecassoside (Madec), a triterpenoid saponin present in Centella asiatica herbs, has a potent anti-inflammatory effect. In the present study, Madec exerted an obvious therapeutic effect in reversing the histological lesions in adjuvant-induced arthritis (AIA) rats. To recognize the anti-rheumatoid potentials of Madec, we further investigated whether Madec interfered with FLS invasion and metalloproteinase (MMP) expression. In cultures of primary FLS isolated from the AIA rats, Madec (10 and 30 μmol·L) was proven to considerably inhibit migration and invasion of FLS induced by interleukin 1β (IL-1β), but exhibiting no obvious effect on cell proliferation. Madec repressed IL-1β-triggered FLS invasion by prohibiting the expression of MMP-13. Additionally, Madec suppressed MMP-13 transcription via inhibiting the MMP-13 promoter-binding activity of NF-κB. Our results further showed that Madec down-regulated the translocation and phosphorylation of NF-κB as demonstrated by Western blotting and immunofluorescence assays. In conclusion, our results suggest that Madec exerts anti-RA activity via inhibiting the NF-κB/MMP-13 pathway.

Anti-rheumatoid arthritic effect of volatile components in notopterygium incisum in rats via anti-inflammatory and anti-angiogenic activities / 中国天然药物

Notopterygium incisum (QH) has been used for the treatment of rheumatoid arthritis (RA), and volatile oils may be its mainly bioactive constituents. The present study was designed to analyze the volatile compounds in QH and to determine the anti-arthritic capacity of Notopterygium volatile oils and the potential mechanism of action. The volatile compounds analysis was conducted by GC-MS. The anti-arthritic capacity test of the volatile oils was conducted on adjuvant-induced arthritis (AIA) rats. The anti-inflammatory property was tested in NO release model in RAW 264.7 cells. Endothelial cells were used to evaluate the anti-proliferative and anti-tube formative effects. 70 compounds were analyzed by GC-MS in the volatile oils. Notopterygium volatile oils weakened the rat AIA in a dose-dependent manner (2, 4, and 8 g crude drug/kg). The NO production by RAW 264.7 was decreased by more than 50% in Notopterygium volatile oils (5, 15, and 45 μg·mL) pretreated groups. Notopterygium volatile oils also inhibited EAhy926 cell proliferation and further delayed EAhy926 cell capillary tube formation in a concentration-dependent manner. The anti-NO productive, anti-proliferative, and anti-tube formative effects of Notopterygium volatile oils strongly suggested that the therapeutic effect of QH in AIA might be related to the potent anti-inflammatory and anti-angiogenic capacities of the volatile oils.

Madecassoside impedes invasion of rheumatoid fibroblast-like synoviocyte from adjuvant arthritis rats via inhibition of NF-κB-mediated matrix metalloproteinase-13 expression / 中国天然药物

Fibroblast-like synoviocytes (FLS) play a pivotal role in Rheumatoid arthritis (RA) pathogenesis through aggressive migration and invasion. Madecassoside (Madec), a triterpenoid saponin present in Centella asiatica herbs, has a potent anti-inflammatory effect. In the present study, Madec exerted an obvious therapeutic effect in reversing the histological lesions in adjuvant-induced arthritis (AIA) rats. To recognize the anti-rheumatoid potentials of Madec, we further investigated whether Madec interfered with FLS invasion and metalloproteinase (MMP) expression. In cultures of primary FLS isolated from the AIA rats, Madec (10 and 30 μmol·L) was proven to considerably inhibit migration and invasion of FLS induced by interleukin 1β (IL-1β), but exhibiting no obvious effect on cell proliferation. Madec repressed IL-1β-triggered FLS invasion by prohibiting the expression of MMP-13. Additionally, Madec suppressed MMP-13 transcription via inhibiting the MMP-13 promoter-binding activity of NF-κB. Our results further showed that Madec down-regulated the translocation and phosphorylation of NF-κB as demonstrated by Western blotting and immunofluorescence assays. In conclusion, our results suggest that Madec exerts anti-RA activity via inhibiting the NF-κB/MMP-13 pathway.

Gentiopicroside,originated from Gentiana macrophylla Pall,possesses anti-arthritic efficacy in adjuvant-induced arthritis rats / 中国药理学与毒理学杂志

OBJECTIVE This work aimed to investigate the anti-rheumatoid arthritic effect of gentio-picroside from Gentiana macrophylla Pall using an animal model of adjuvant induced arthritis. METH-ODS Adjuvant arthritis was induced in fifty SD male rats,which were randomly divided into five groups (n=10):control(0.5% CMC-Na)group,AIA(rats with CFA)group,dexamethasone(1 mg·kg-1)group, gentiopicroside(50 mg·kg-1)group,and gentiopicroside(100 mg·kg-1)group.Rats were administered intragastrically with drugs or CMC-Na once a day for a period of 2 weeks.Paw swelling,arthritic index, histological changes were assessed to evaluate the anti-arthritic effect.Weight growth,spleen and thymus indexes were also investigated in.RESULTS Gentiopicroside at dose of 100 mg·kg-1significantly inhibited the secondary paw swelling(P<0.05)and arthritis index(P<0.05),decreased synovial inflammatory infil-tration, synovial hyperplasia and bone erosion. Furthermore, gentiopicroside showed no immunosup-pressive adverse effects in body weight, index of spleen and thyums compared with dexamethasone administration (P<0.05, P<0.01). CONCLUSION Gentiopicroside possessed anti-arthritic efficacy in AIA rats without immunosuppressive effects.

Anti-rheumatoid arthritic effect of volatile components in notopterygium incisum in rats via anti-inflammatory and anti-angiogenic activities / 中国天然药物

Notopterygium incisum (QH) has been used for the treatment of rheumatoid arthritis (RA), and volatile oils may be its mainly bioactive constituents. The present study was designed to analyze the volatile compounds in QH and to determine the anti-arthritic capacity of Notopterygium volatile oils and the potential mechanism of action. The volatile compounds analysis was conducted by GC-MS. The anti-arthritic capacity test of the volatile oils was conducted on adjuvant-induced arthritis (AIA) rats. The anti-inflammatory property was tested in NO release model in RAW 264.7 cells. Endothelial cells were used to evaluate the anti-proliferative and anti-tube formative effects. 70 compounds were analyzed by GC-MS in the volatile oils. Notopterygium volatile oils weakened the rat AIA in a dose-dependent manner (2, 4, and 8 g crude drug/kg). The NO production by RAW 264.7 was decreased by more than 50% in Notopterygium volatile oils (5, 15, and 45 μg·mL) pretreated groups. Notopterygium volatile oils also inhibited EAhy926 cell proliferation and further delayed EAhy926 cell capillary tube formation in a concentration-dependent manner. The anti-NO productive, anti-proliferative, and anti-tube formative effects of Notopterygium volatile oils strongly suggested that the therapeutic effect of QH in AIA might be related to the potent anti-inflammatory and anti-angiogenic capacities of the volatile oils.

Programmed cell death 5 transgenic mice attenuates adjuvant induced arthritis by 2 modifying the T lymphocytes balance

BACKGROUND: Programmed cell death 5 (PDCD5) is an apoptosis-related gene cloned from TF-1 cells whose primary biological functions are to promote apoptosis and immune regulation. The effects and mechanisms exerted by key mediators of arthritic inflammation remain unclear in PDCD5 transgenic (PDCD5 tg) mice. RESULTS: In the current study, PDCD5 tg mice inhibited the progression of adjuvant-induced arthritis, specifically decreasing clinical signs and histological damage, compared with arthritis control mice. Additionally, the ratio of CD4+IFN-γ+ cells (Th1) and CD4+IL-17A+ cells (Th17), as well as the mRNA expression of the pro-inflammatory mediators IFN-γ, IL-6, IL-17A and TNF-α, were decreased in PDCD5 tg mice, while CD4+CD25+Foxp3+ regulatory T (Treg) cells and the anti-inflammatory mediators IL-4 and IL-10 were increased. Furthermore, PDCD5 tg mice demonstrated reduced serum levels of IFN-γ, IL-6, IL-17A and TNF-α and increased levels of IL-4. CONCLUSIONS: Based on our data, PDCD5 exerts anti-inflammatory effects by modifying the T lymphocytes balance, inhibiting the production of pro-inflammatory mediators and promoting the secretion of anti-inflammatory cytokines, validating PDCD5 protein as a possible treatment for RA.

Tujia ethnomedicine Xuetong suppresses onset and progression of adjuvant-induced arthritis in rats / 中国药理学通报

Aim Kadsuraheteroclita ( Roxb ) Craib ( Schizandraceae) is a medicinal plant termed Xuetong in Chinese Tujia ethnomedicine. Xuetong possesses therapeutic effects of, in the terms of Chinese medical theories, reinforcing vital energy, promoting blood cir-culation, expelling wind-evil, and removing damp-e-vil, and has been long used for the prevention and treatment of rheumatic and arthritic diseases, especial-ly in the southern China. The HPLC analysis has iden-tified that the ethanol extract of Xuetong contains large-ly biologically active lignans and triterpenoids. Our previous studies have shown that KHS exhibits very fa-vorable safety profile and potent anti-inflammatory and analgesic activities. In the present study, we investiga-ted anti-arthritic effects and the possible mechanisms of Xuetongon adjuvant-induced arthritis ( AIA ) in rats. Methods AIA was established in male Sprague-Daw-ley ( SD ) rats as described previously, and animals were daily treated by gavage with Xuetong ethanol ex-tract ( 1. 0 g · kg-1 ) or vehicle ( 0. 3% CMC-Na ) throughout the 30-day experiment. The incidence and severity of arthritis were evaluated using clinical pa-rameters. On day 30, bone destruction of the arthritic joints was assessed by computed tomography( CT) and histopathological analyses. The serum levels of pro-in-flammatory cytokines TNF-α, IL-1β, and IL-6 were measured by ELISA. Results Treatment with 1. 0 g/kg Xuetong significantly inhibited the onset and pro-gression of AIA. The vehicle-treated rats all developed severe arthritis, while the incidence of AIA in the Xue-tong-treated rats was as low as 55%( P=0. 035 ) . The Xuetong -treated rats exhibited 1. 8 to 2. 3 fold reduc-tion of paw swelling, and gained 10 to 20% more body weight than the vehicle-treated AIA rats throughout the experiment. CT and histopathological examinations re-vealed that Xuetong markedly protected AIA rats from cartilage and bone destruction of joints. Moreover, the serum levels of TNF-α, IL-1β, and IL-6 were signifi-cantly decreased in the Xuetong-treated rats than the vehicle-treated AIA rats. Conclusions These data strongly support the clinical use of Xuetong for rheu-matic and arthritic diseases, and suggest that Xuetong is a valuable candidate for further investigation to be a new anti-arthritic drug with favorable safety profile.

Pharmacodynamic Effects of Volatile Oil from Homalomena occulta on Adjuvant-induced Arthritis Model Rats and Its Mechanism Study / 中国药房

OBJECTIVE:To study the pharmacodynamic effects of volatile oil from Homalomena occulta on adjuvant-induced arthritis(AA)model rats and its mechanism. METHODS:60 rats were randomly divided into normal control group(0.5%polysor-bate 80),model control group (0.5% polysorbate 80),positive control group (Tripterygium glycosides tablets,10 mg/kg), high-dose,middle-dose and low-dose(0.08,0.04 and 0.02 ml/kg)groups of volatile oil from H. occulta. Except for normal control group,other groups were given complete Freund's adjuvant subcutaneously via right rear foot plantar to induce AA model,and giv-en relevant medicine intragastrically for 25 days,once a day,since modeling. The articular swelling degree,immune organ (thy-mus gland and spleen) index,pathological change,the contents of IL-1β and TNF-α in serum were detected. RESULTS:Com-pared with normal control group,the primary and secondary articular swelling degree and thymus gland index of rats and the serum contents of IL-1β and TNF-α increased in model control group,while spleen index decreased(P<0.05 or P<0.01);obvious tissue swelling,large amount of neutrophile granulocyte,leukomonocyte and macrophage infiltrating joint surrounding tissue,the prolifer-ation of synovial cells and obvious osteoarthritic lesion were observed in podarthrum. Compared with model control group,the pri-mary and secondary articular swelling degree and the serum content of IL-1β decreased in the volatile oil from H. occulta groups;thymus gland index increased in middle-dose and low-dose groups of the volatile oil from H. occulta;the content of TNF-α de-creased in high-dose group of the volatile oil from H. occulta(P<0.05 or P<0.01). The inflammatory cell infiltration of joint sur-rounding tissue relieved in treatment groups,synovial cells proliferation was not obvious and synovial cells morphology was im-proved. CONCLUSIONS:The volatile oil from H. occulta has the pharmacodynamic effects on AA in rat,and its mechanism might be related to the serum content reduction of IL-1β and TNF-α.

Effect of Acanthopanax trifo-liatus Alcohol Extract on the Contents of Inflammatory Factors in Serum of Adjuvant-induced Arthritis Rat / 中国药房

OBJECTIVE:To study the effect of Yi medicine Acanthopanax trifo-liatus alcohol extract on the contents of IL-1β, IL-6,IL-10,NO,TNF-αand PGE2 in serum of adjuvant-induced arthritis(AA)rats. METHODS:60 SD rats were randomly divid-ed into normal control group (distilled water),model group (distilled water),positive control group (nimesulide,0.03 g/kg), A. trifo-liatus high-dose,medium-dose and low-dose groups [7,3.5,1.75 g(medicinal material)/kg]. Except for normal control group,other groups were given 0.1 ml Freund’s complete adjuvant on posterior right toe to induce AA model. After modeling, those groups were given relevant medicine intragastrically,once a day. The contents of IL-1β,IL-6,IL-10,TNF-α,NO,PGE2 in rat serum were measured 30 days later. The pathological morphology of the rat ankle was also observed. RESULTS:Compared with normal control group,the contents of IL-1β,IL-6,TNF-α,NO and PGE2 in serum increased in model group while the content of IL-10 decreased,with statistical significance (P<0.01);compared with model group,the contents of IL-1β,IL-6,TNF-α,NO and PGE2 in serum decreased in A. trifo-liatus high-dose groups while the content of IL-10 decreased,with statistical significance (P<0.05). The pathology results showed that part of the slice synoviocytes mild thickening,fibrous tissue mild hyperplasia,in-flammatory cells infiltration at varying degrees,macrophages mild hyperplasia in A. trifo-liatus groups. Those changes were not found in normal control group. CONCLUSIONS:A. trifo-liatus alcohol extract has an obvious anti-inflammatory action to AA mod-el rat,by a mechanism of decreasing the contents of IL-1β,IL-6,TNF-α,NO and PGE2,and increasing the content of IL-10.

Evaluation of antinociceptive and anti-inflammatory activities of extract and fractions of Eugenia jambolana root bark and isolation of phytoconstituents

Eugenia jambolana Lam., Myrtaceae, is a widely distributed and traditionally well known plant in India. The root bark of the plant was extracted with ethanol and then successively fractionated into petroleum ether fraction, chloroform fraction, n-butanol fraction and methanol fraction. The extract and fractions of the plant material were evaluated for the antinociceptive activity by acetic acid-induced writhing test and formalin-induced nociception test, and anti-inflammatory activity was screened by carrageenan-induced rat paw edema, cotton pellet induced granuloma formation and adjuvant induced arthritis in rat models. The test materials showed the antinociceptive and anti-inflammatory effect in dose dependent manner and the petroleum ether fraction was found to be most potent among the test materials. At 400 mg/kg b.w., p.o. dose petroleum ether fraction significantly inhibited 54.28% writhing response and 73.77% formalin induced nociception in mice. The fraction with same dose showed significant 79.31% inhibition of carrageenan-induced rat paw edema, 57.78% anti-proliferative effect and 77.93% inhibition of adjuvant induced arthritis. The bioactive petroleum ether fraction was then subjected to column chromatography which led to isolate three compounds, namely, β-sitosterol, stigmasterol and lupeol. These compounds were characterized and identified by IR, ¹H NMR, 13C NMR and mass spectroscopy.

Anti-inflammatory effects of ramipril in experimentally induced rheumatoid arthritis.

Objective: To evaluate anti-inflammatory effects of ramipril in experimentally induced rheumatoid arthritis (RA).Materials and Methods: Adjuvant induced arthritis model is used in this study.Albino-Wistar rats of either sex were used. Arthritis was induced by single intradermal injection of Freund’s complete adjuvant (FCA) suspended in oil inplantar region of right hind paw. Rats were divided in to three groups (n=8) namely disease control, standard and test group. Drug treatment was carried out for 21 days. Effect of test drug on acute inflammatory phase was evaluated on day 5 by assessing right hind paw edema. After 21 days animals were sacrificed and evaluated for left hind paw edema, weight changes, histo-pathological synovitis grading in left hind limbs and secondary lesion score.Results: Results showed that ramiprilsignificantly reduced right hind paw edema on day 5 (p<0.05). Ramipril also showed statistically significant weight gain (p<0.05), reduction of histo-pathological synovitis grade (p<0.05) as well as secondary lesion score (p<0.05).Conclusion:Our study suggests that ramiprilmay be used as an adjuvant anti-inflammatory agent in patients with RA. However this speculation needs to be confirmed clinically.

Expression of EAAC1 in dorsal root ganglion in rats with inflammatory pain-morphine tolerance / 临床麻醉学杂志

Objective To explore the role of excitatory amino acid carrier 1 (EAAC1)in dorsal root ganglion (DRG) in the mechanism of developing morphine tolerance. Methods Thirty male SD rats were implanted intrathecal catheters and randomized into 6 groups with 5 rats each. The rats of 4 groups were made into the model of adjuvant-induced arthritis in the left hind limb and were administered intrathecally, morphine 10 μg(group M_(10)), morphine 20μg(group M_(20)), morphine 20 μg plus naloxone 10 μg(group MN) ,or saline(group C) respectively. The other 2 groups without were administered intrathecally saline (group C_0) or morphine 20 μg (group M0). The drugs were administered twice daily for 7 days. Mechanical withdrawl threshold(MWT) of the left hind limb was examined to evaluate the behavior. Immunohistochemistry was used to detect the expression of EAAC1 in the left L_(3-4) and L_(4-5) DRG. Results Morphine tolerance was formmed stably in the arthritis rats of group M_(10) and group M_(20) after administering morphine for 7 days. The expression of EAAC1 in DRG was downregulated. Conclusion DRG EAAC1 may be involved in the mechanism of developing morphine tolerance in rats with inflammatory pain.