RESUMEN
Dehydroevodiamine (DHE) is an effective natural active substance extracted from Euodiae Fructus, which is a widely used herbal drug in traditional Chinese medicine. The focus of this study was to test the possibility of using DHE in the treatment of rheumatoid arthritis (RA) diseases. A rat model of adjuvant-induced arthritis (AIA) was generated using Complete Freund's Adjuvant (CFA). Body weight changes, arthritis scores, ankle pathology, tumor necrosis factor-alpha (TNF-α), interleukin-1ß(IL-1ß), interleukin-6 (IL-6), and interleukin-17 (IL-17) secretion, as well as matrix metalloproteinase (MMP) expression in joint tissue, were measured as indicators of viability of DHE medicated AIA rats. Human fibroblast-like synoviocytes (MH7A cells) were connected to check these impacts. The results confirmed that DHE administration had an excellent therapeutic impact on the AIA rat model, substantially relieving joint swelling, inhibiting synovial pannus hyperplasia, and decreasing joint scores. In addition, the serum enzyme-linked immunosorbent assay (ELISA) showed that DHE treatment reduced the expression of pro-inflammatory factors in AIA rats. The immunohistochemical results showed that DHE treatment could reduce the synthesis of MMPs such as matrix metalloproteinase-1(MMP-1) and matrix metalloproteinase-3 (MMP-3) in the ankle tissue of AIA rats. In vitro, DHE inhibited cell proliferation, mRNA transcription, protein synthesis of proinflammatory factors such as IL-1ßand IL-6, and matrix metalloproteinases such as MMP-1 and MMP-3. Furthermore, DHE inhibited the phosphorylation levels of p38, JNK, and ERK proteins in TNF-α-treated MH7A cells.This work assessed the effect of DHE in AIA rats and revealed its mechanism in vitro.
Asunto(s)
Alcaloides/administración & dosificación , Antiinflamatorios/administración & dosificación , Artritis Experimental/tratamiento farmacológico , Adyuvante de Freund/efectos adversos , Sinoviocitos/citología , Alcaloides/farmacología , Animales , Antiinflamatorios/farmacología , Artritis Experimental/inducido químicamente , Artritis Experimental/genética , Artritis Experimental/inmunología , Peso Corporal/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citocinas/genética , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Metaloproteinasas de la Matriz/metabolismo , Ratas , Sinoviocitos/efectos de los fármacos , Sinoviocitos/inmunologíaRESUMEN
The present study aimed to investigate the antiarthritic effect of physcion 8Oßglucopyranoside (POGD) and its possible mechanisms. The antiproliferative effects of POGD on MH7A cells were detected using a CCK8 assay, and the release of proinflammatory cytokines, interleukin (IL)1ß, IL6, IL8, IL12 and IL17A, were determined by ELISA. A type II collageninduced arthritis (CIA) rat model was established to evaluate the antiarthritic effect of POGD in vivo. The paw volumes, arthritis indices and serum levels of tumor necrosis factor (TNF)α, IL1ß, IL6, IL8, IL17A were determined by ELISA. The mRNA expression levels of matrix metalloproteinase (MMP)2, MMP3, MMP9, vascular endothelial growth factor and cyclooxygenase2 were determined by reverse transcriptionquantitative polymerase chain reaction analysis, and the expression levels of transforming growth factor (TGF)ß1, small mothers against decapentaplegic (Smad)4, Smad7, cJun Nterminal kinase (JNK), phosphorylated (p)JNK, pP38, P38, pextracellular signalregulated kinase (ERK)1/2, ERK1/2, nuclear factor (NF)κB p65 in the nucleus (N), cytosolic NFκB p65 (C), and inhibitor of NFκB (IκB) were determined by western blot analysis. The results indicated that POGD significantly inhibited MH7A cell growth. POGD markedly inhibited paw swelling and the arthritis indices of the CIA rats, and POGD may also inhibit the release of proinflammatory cytokines. Furthermore, POGD downregulated the expression levels of TGFß1, Smad4, NFκB p65 (N), p38, pp38, pERK1/2, JNK, pJNK, TGFß1, Smad4, pJNK, JNK, pP38, P38, pERK1/2, ERK1/2 and NFκB p65 (N), and upregulated the Smad7, NFκB p65 (C) and IκB in TNFα induced MH7A cells. In conclusion, the results suggested that POGD is a promising potential antiinflammatory drug, and that POGD may decrease the expression of proinflammatory cytokines and mediators via inhibiting the TGFß/NFκB/mitogenactivated protein kinase pathways.
Asunto(s)
Antiinflamatorios/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Artritis Reumatoide/tratamiento farmacológico , Emodina/análogos & derivados , Glucósidos/uso terapéutico , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Sinoviocitos/efectos de los fármacos , Factor de Crecimiento Transformador beta/inmunología , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Artritis Experimental/inmunología , Artritis Experimental/patología , Artritis Reumatoide/inmunología , Artritis Reumatoide/patología , Línea Celular , Proliferación Celular/efectos de los fármacos , Emodina/química , Emodina/aislamiento & purificación , Emodina/uso terapéutico , Fallopia japonica/química , Femenino , Glucósidos/química , Glucósidos/aislamiento & purificación , Interleucinas/inmunología , Masculino , Ratas , Transducción de Señal/efectos de los fármacos , Sinoviocitos/citología , Sinoviocitos/inmunología , Sinoviocitos/patologíaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Aconiti Sinomontani Radix is frequently used in the treatment of Bi syndrome in traditional Chinese medicine. Several reports indicate that Aconiti Sinomontani Radix has therapeutic effects for rheumatoid arthritis (RA). However, the cellular mode of action is still unclear. To investigate the effect of alkaloid extracts of Aconiti Sinomontani Radix on proliferation and migration of human synovial sarcoma SW982 cells as well as the molecular mechanism underlying. MATERIALS AND METHODS: SW982 cells were examined for proliferation by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method. Wound scratch assays were performed to assess the migrated rate of SW982 cells. Quantitative real-time PCR was used to measure the mRNA expression levels of Wnt5a, Runx2, MMP3, and Bmp2. Western blotting was used to measure the phosphorylated levels of JNK and NF-κB as well as the expression of MMP3. RESULTS: The alkaloid extract from Aconiti Sinomontani Radix (MQA) and MQB, which removed lappaconitine from MQA significantly inhibited the proliferation of SW982 in a dose-dependent manner. The proliferation inhibitory effect of MQB was more potent. Incubation with 10µg/ml MQB for 12, 24, and 36h inhibited the migration of SW982 cells by 83%, 58%, and 42%, respectively. Treatment with different concentrations of MQB for 24h inhibited mRNA expression of Wnt5a, Runx2, and MMP3, but Bmp2 mRNA expression was elevated by MQB. Further, MQB inhibited phosphorylation of JNK and NF-κB p65 as well as MMP3 expression by Western blotting analysis. CONCLUSION: The results showed that MQB inhibited proliferation and migration of SW982 cells possibly through suppressing Wnt5a-mediated JNK and NF-κB pathways. These results indicated that MQB might be an active extract of Aconiti Sinomontani Radix for targeting fibroblast-like synoviocytes (FLS) and be potential for RA therapy.
Asunto(s)
Aconitum/química , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Extractos Vegetales/farmacología , Sinoviocitos/citología , Sinoviocitos/efectos de los fármacos , Proteína Morfogenética Ósea 2/biosíntesis , Línea Celular , Ensayos de Migración Celular , Subunidad alfa 1 del Factor de Unión al Sitio Principal/biosíntesis , Relación Dosis-Respuesta a Droga , Fibroblastos/citología , Expresión Génica/efectos de los fármacos , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Metaloproteinasa 3 de la Matriz/biosíntesis , FN-kappa B/metabolismo , Fosforilación/efectos de los fármacos , Proteína Wnt-5a/biosíntesisRESUMEN
Three guaiane-type sesquiterpenoids, dysodensiols G-I (1-3), together with five known sesquiterpenoids (4-8) were isolated from the stems of Fissistigma oldhamii (Hemsl.) Merr. Compound 1 represents the first example of an ene(6 â 5)-abeo-14-norguaiane sesquiterpenoid derived from natural products. Their structures were elucidated by a combination of 1D and 2D NMR and MS spectra. The absolute configuration of 2 was determined by an X-ray crystallographic analysis. The inhibitory effect of all compounds on the proliferation of primary synovial cells was evaluated. Compound 3 showed a potent inhibitory effect on the proliferation of synoviocytes with an IC50 value of 1.0 µM.