RESUMEN
BACKGROUND: ChondroT, a new herbal medication, consists of the water extracts of Osterici Radix, Lonicerae Folium, Angelicae Gigantis Radix, Clematidis Radix, and Phellodendri Cortex (6:4:4:4:3). We previously reported that ChondroT showed significant anti-arthritis and anti-inflammatory effects. METHODS: This study was designed to evaluate the effect of ChondroT on hyperuricemia. First, the effect of ChondroT was evaluated on xanthine oxidase (XOD) activity in vitro. The anti-hyperuricemic effect of ChondroT was also studied in potassium oxonate (PO)-induced hyperuricemic model mice. Uric acid (UA) and XOD were evaluated in the serum, urine, and liver of the mice. In addition, we measured serum creatinine (Cr) and blood urea nitrogen (BUN) levels as well as mRNA expression of the mouse urate transporter 1 (mURAT1) to evaluate kidney function and urate excretion in hyperuricemic mice. RESULTS: ChondroT showed in vitro XOD inhibitory activity in a dose-dependent manner (P < 0.05). We demonstrated that ChondroT (37.5, 75 and 150 mg/kg) significantly reduced serum UA (P < 0.01 and P < 0.001, respectively), and upregulated urinary UA (P < 0.001, respectively) in PO-induced hyperuricemic mice. In addition, ChondroT (75 and 150 mg/kg) significantly reduced Cr (P < 0.05 and P < 0.01, respectively), BUN (P < 0.05 and P < 0.001, respectively), GOT (P < 0.05 and P < 0.01, respectively), and GPT (P > 0.05 and P < 0.05, respectively) levels in PO-induced hyperuricemic mice. ChondroT (75 and 150 mg/kg) also significantly downregulated serum (P < 0.05) and liver (P < 0.05) XOD activity. Compared to the hyperuricemic mice, the ChondroT (37.5, 75, and 150 mg/kg)-treated mice showed decreased mURAT1 protein expression level. CONCLUSION: ChondroT displayed anti-hyperuricemic effects by regulating XOD activity and kidney mURAT1.
Asunto(s)
Regulación hacia Abajo/efectos de los fármacos , Medicamentos Herbarios Chinos/administración & dosificación , Hiperuricemia/tratamiento farmacológico , Transportadores de Anión Orgánico/genética , Ácido Oxónico/efectos adversos , Xantina Oxidasa/genética , Animales , Creatinina/sangre , Evaluación Preclínica de Medicamentos , Humanos , Hiperuricemia/inducido químicamente , Hiperuricemia/genética , Hiperuricemia/metabolismo , Riñón/efectos de los fármacos , Riñón/fisiopatología , Masculino , Ratones , Ratones Endogámicos ICR , Transportadores de Anión Orgánico/metabolismo , Ácido Úrico/sangre , Xantina Oxidasa/sangreRESUMEN
Toll-like receptor (TLR) ligands are being developed for use as vaccine adjuvants and as immunomodulators because of their ability to stimulate innate and adaptive immune responses. Flagellin, a TLR5 ligand, was reported to show potent mucosal vaccine adjuvant activity. To identify ligands that potentiate the adjuvant activity of flagellin, we screened a plant library using HEK293T cells transiently cotransfected with phTLR5 and pNF- κ B-SEAP plasmids. The 90% EtOH extract from Croton tiglium showed significant NF- κ B transactivation in a TLR5-independent manner along with the increase of a flagellin activity. We have studied to characterize an active component from Croton tiglium and to elucidate the action mechanisms. Phorbol 12-myristate 13-acetate (PMA) was isolated as an active component of Croton tiglium by activity-guided fractionation, column chromatography, HPLC, NMR, and MS. PMA at a range of nM induced PKC-dependent NF- κ B activation and IL-8 production in both TLR5- and TLR5+ assay systems. In in vivo mouse vaccination model, PMA induced antigen-specific IgG and IgA antibody responses and increased IL-12 production corresponding to T cell responses in spleen lymphocytes. These results suggest that PMA would serve as an efficacious mucosal vaccine adjuvant.
Asunto(s)
Adyuvantes Inmunológicos/química , Transducción de Señal , Acetato de Tetradecanoilforbol/química , Receptores Toll-Like/metabolismo , Animales , Células CACO-2 , Núcleo Celular/metabolismo , Croton/química , Citosol/metabolismo , Flagelina/química , Células HEK293 , Humanos , Interleucina-8/metabolismo , Ligandos , Linfocitos/citología , Ratones , Ratones Endogámicos BALB C , FN-kappa B/metabolismo , Extractos Vegetales/química , Bazo/citología , Linfocitos T/inmunología , Receptor Toll-Like 5/metabolismo , Activación Transcripcional , VacunasRESUMEN
Vibrio vulnificus, a good model organism of bacterial septicemia, causes fatal septicemia manifesting a fulminating course and a high mortality rate within days. In order to identify new natural substances preventing V. vulnificus infection, a plant library was screened for inhibiting cytotoxicity to host cells by using Trypan blue staining and LDH assay. We found that Polygoni Cuspidati Radix potently suppressed the acute death of HeLa and RAW264.7 cells in a dose dependent manner. Further studies revealed that Polygoni Cuspidati Radix inhibited V. vulnificus growth and survival in HI broth and seawater, respectively. We confirmed that Polygoni Cuspidati Radix contained high level of emodin by thin layer chromatography (TLC). Emodin showed direct antibacterial activity against V. vulnificus. In addition, emodin prevented the morphologic damages and acute death of HeLa cells caused from V. vulnificus. The safety of Polygoni Cuspidati Radix and emodin to host cells was confirmed by MTT assay. Polygoni Cuspidati Radix and emodin protected mice from V. vulnificus infection.
Asunto(s)
Emodina/farmacología , Fallopia japonica/química , Extractos Vegetales/farmacología , Vibriosis/tratamiento farmacológico , Vibrio vulnificus/efectos de los fármacos , Animales , Técnicas Bacteriológicas , Bioensayo , Línea Celular , Supervivencia Celular , Medios de Cultivo , Emodina/uso terapéutico , Células HeLa , Humanos , Ratones , Extractos Vegetales/química , Extractos Vegetales/uso terapéutico , Agua de Mar/microbiología , Vibriosis/microbiología , Vibriosis/mortalidad , Vibrio vulnificus/crecimiento & desarrollo , Vibrio vulnificus/patogenicidadRESUMEN
Vibrio vulnificus is a human pathogen causing a rapidly progressing fatal septicemia. We have previously reported that a V. vulnificus large toxin RtxA1 causes programmed necrotic cell death through calcium-mediated mitochondrial dysfunction. Here we developed a live attenuated vaccine strain (CMM781) having deletions in three genes encoding major virulence factors: RTX cytotoxin (rtxA1), hemolysin/cytolysin (vvhA) and metalloprotease (vvpE) of a clinical isolate strain CMCP6. The CMM781 strain showed significant attenuation in cytotoxicity and mouse lethality. The safety of CMM781 was also confirmed by measuring the transepithelial electric resistance of Caco-2 cell monolayers. Intragastric immunization of mice with the live attenuated V. vulnificus strain resulted in induction of systemic and mucosal antibodies specific to the pathogen. Moreover, the vaccinated mice were protected from challenges with high doses of the virulent strain through various injection routes. These results suggest that CMM781 appears to be a safe and effective vaccine candidate that would provide significant protection against V. vulnificus infection.
Asunto(s)
Vacunas Bacterianas/efectos adversos , Vacunas Bacterianas/inmunología , Vibriosis/prevención & control , Vibrio vulnificus/inmunología , Factores de Virulencia/genética , Animales , Anticuerpos Antibacterianos/sangre , Toxinas Bacterianas/genética , Vacunas Bacterianas/administración & dosificación , Células CACO-2 , Citotoxinas/genética , Células HeLa , Proteínas Hemolisinas/genética , Humanos , Dosificación Letal Mediana , Metaloproteasas/deficiencia , Metaloproteasas/genética , Ratones , Mutación , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/efectos adversos , Vacunas Atenuadas/genética , Vacunas Atenuadas/inmunología , Vibriosis/inmunología , Vibriosis/microbiología , Vibrio vulnificus/genéticaRESUMEN
Vibrio vulnificus (V. vulnificus) is a useful model for bacterial septicemia as the bacterial infection generates a wide pathogenic spectrum in addition to a high mortality rate. This study was aimed at investigating the effects of genistein on the growth, cytotoxicity, adhesion, and the mouse mortality caused by V. vulnificus. The results of our study indicated that genistein (50 or 100 mg/L) effectively minimized the morphologic damages and inhibited acute death of HeLa cells by V. vulnificus. Strikingly, genistein significantly inhibited the adhesion of V. vulnificus to HeLa cells. This report confirmed that genistein showed bacteriostatic activity against V. vulnificus, but it did not exhibit any bactericidal activity. Nevertheless, genistein (20 mg/kg) effectively decreased CD-1 mice mortality caused by V. vulnificus infection.
Asunto(s)
Adhesión Bacteriana/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Genisteína/farmacología , Genisteína/uso terapéutico , Vibriosis/tratamiento farmacológico , Vibrio vulnificus/efectos de los fármacos , Animales , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Células HeLa , Humanos , Ratones , Ratones Endogámicos , Pruebas de Sensibilidad Microbiana/métodos , Vibriosis/mortalidad , Vibrio vulnificus/crecimiento & desarrollo , Vibrio vulnificus/patogenicidadRESUMEN
Host-parasite contact is a prerequisite for the acute cytotoxicity of Vibrio vulnificus, which is mediated primarily by RtxA1, a repeat in toxin (RTX) toxin. We found that resveratrol (at 10 or 30 microM), a natural polyphenol, protected HeLa cells from V. vulnificus cytotoxicity. To further characterize the underlying mechanism, the effect of resveratrol was investigated at the level of the host-microbe interactions. We studied the effects of resveratrol on adhesion, motility, cytotoxicity, and RtxA1 toxin expression of V. vulnificus. In addition, the effect of resveratrol on mouse mortality caused by V. vulnificus was investigated. Resveratrol inhibited V. vulnificus motility and the microbe adhesion to host cells, critical virulence traits for many bacteria. Resveratrol also down-regulated the expression of RtxA1 toxin at the transcriptional level and thereby protected the host cells from becoming rounded and damaged. In addition, resveratrol (20mg/kg) protected CD-1 mice from V. vulnificus infection. Taken together, these results suggest that resveratrol, a modulator of host-microbe interactions, has potential for development as a new paradigm drug to treat infectious diseases.