RESUMEN
BACKGROUND: The herbal formula (Sahatsatara, STF), the Thai traditional poly-herbal recipe, has been used for treatment of muscle pain, anti-flatulence and numbness on hands and feet, with the caution when used in hypertensive patients. However, there is no scientific evidence to prove its effects on cardiovascular system. Piperine is the proposed major active compound in STF. It is shown to have antihypertensive effect in the L-NAME-induced endothelial dysfunction rats. PURPOSE: This study investigated the pharmacokinetics, mechanism of action, as well as the hemodynamic and vasoactive effect and toxicity of STF and piperine using spontaneously hypertensive rats (SHR) and normal Wistar rats (NWR). METHODS: The amount of piperine in STF was measured by ultra performance liquid chromatography (UPLC). SHR and NWR were gavaged with piperine (50mg/kg/day) or STF (100, 300, or 1000mg/kg/day) alone or together with L-NAME (in drinking water) for 28 days. Hemodynamic effects were monitored by noninvasive tail cuff every 7 days. Vasorelaxation effect on the thoracic aorta in organ chamber was observed through force transducer at the end of the experiment. Biochemical parameters for kidney and liver toxicity were measured. In addition, pharmacokinetic study was performed using non-compartment analysis. RESULTS: The amount of piperine in STF was 1.29%w/w. Both STF and piperine did not affect blood pressure and heart rate in both SHR and NWR. Interestingly, STF and piperine increased acetylcholine-induced vasorelaxation of isolated thoracic aorta and have vascoluprotective effect in nitric oxide (NO) impaired rats. No liver or kidney toxicity was found in this study. Non-compartment pharmacokinetic analysis showed that the time to reach maximum concentration (Tmax) of plasma piperine after administration of piperine and STF were 3.9 and 1.7h, respectively. This result suggested that piperine in the recipe had better absorption than the pure standard piperine. CONCLUSIONS: STF had no effect on blood pressure in both SHR and NWR. However, it was able to relax isolated thoracic aorta and had the potential for vasculoprotective effect in hypertensive and NO impaired condition. The effects of STF were comparable to those of piperine.
Asunto(s)
Alcaloides/uso terapéutico , Antihipertensivos/uso terapéutico , Benzodioxoles/uso terapéutico , Presión Sanguínea/efectos de los fármacos , Hipertensión/tratamiento farmacológico , Piperidinas/uso terapéutico , Extractos Vegetales/uso terapéutico , Plantas Medicinales/química , Alcamidas Poliinsaturadas/uso terapéutico , Vasodilatación/efectos de los fármacos , Alcaloides/farmacología , Animales , Antihipertensivos/farmacología , Benzodioxoles/farmacología , Masculino , Fitoterapia , Piperidinas/farmacología , Extractos Vegetales/farmacología , Alcamidas Poliinsaturadas/farmacología , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Ratas Wistar , TailandiaRESUMEN
BACKGROUND: A dried root of Aristolochia tagala Cham. (ATC) is often used in Thai traditional medicine as an antipyretic, anti-inflammatory agent, muscle relaxant, appetite-enhancing agent, and analeptic. Homnawakod, an important herbal recipe, originally contains ATC in its formula, however, some Aristolochia species have been reported to cause nephrotoxicity due to aristolochic acid (AA) and its derivatives, resulting in ATC removal from all formulae. Therefore, this study investigates the chemical profiles of ATC, the original (HNK+ATC) and the present Homnawakod Ayurved Siriraj Herbal Formulary™ (HNK), and investigates whether they could cause nephrotoxicity or aggravate LPS-induced organ injuries in vivo. METHODS: HPLC and LC/MS were used for chemical profile study. Male Wistar rats were randomly divided into groups in which the rats were intragastrically administered distilled water (2 groups), ATC (10 or 30 mg/kg), HNK+ATC (540 or 1,620 mg/kg), or HNK (1,590 mg/kg) for 21 days. A positive control group was administered with single dose 100 mg/kg standard AA-I intragastrically at day 1. Serum creatinine and urea were measured at baseline and at 7, 14 and 21 days of the treatment. On day 22, a model of lipopolysaccharide (LPS)-induced endotoxemia was used. One-way and two-way analyses of variance were performed and a P value of less than 0.05 was considered to be significant. RESULTS: The similarity of the HPLC chromatograms of HNK+ATC and HNK could suggest that the qualities of both formulae are nearly the same in terms of chemical profile. The amount of AA-I found in ATC is 0.24%w/w. All experimental groups exhibited similar levels of serum urea at baseline and 7 and 14 days of the treatment. At 21 days, rats received AA exhibited a significant increase in serum urea, whereas the others did not exhibit such toxicity. On day 22, there were no significant changes in LPS-induced renal and liver dysfunction, or LPS-induced mean arterial pressure (MAP) reduction upon administration of ATC, HNK+ATC, HNK or AA-I. CONCLUSIONS: These results suggest that ATC, HNK+ATC or HNK, at the animal dose equivalent to that used in human, do not cause the acute nephrotoxicity in rats and do not aggravate LPS-induced organ injuries even further.
Asunto(s)
Aristolochia/efectos adversos , Ácidos Aristolóquicos/efectos adversos , Endotoxemia/fisiopatología , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Extractos Vegetales/efectos adversos , Índice de Severidad de la Enfermedad , Animales , Aristolochia/química , Ácidos Aristolóquicos/análisis , Ácidos Aristolóquicos/farmacología , Presión Sanguínea/efectos de los fármacos , Enfermedad Hepática Inducida por Sustancias y Drogas/fisiopatología , Cromatografía Líquida de Alta Presión , Endotoxemia/inducido químicamente , Hipotensión/inducido químicamente , Riñón/fisiopatología , Enfermedades Renales/inducido químicamente , Enfermedades Renales/fisiopatología , Lipopolisacáridos , Hígado/fisiopatología , Masculino , Medicina Ayurvédica , Medicina Tradicional de Asia Oriental , Extractos Vegetales/química , Extractos Vegetales/farmacología , Ratas , Ratas Wistar , Urea/sangreRESUMEN
BACKGROUND: Phyllanthus emblica L. (Indian gooseberry, Ma khaam pom) has been an herbal component of Thai traditional recipes proposed to slow down the aging process. A number of methodologies have been employed to investigate the immunological aspects of the so called "anti-aging effects" of P. emblica in a BALB/c mice model. OBJECTIVE: 1) To investigate the immunological efficacy of the anti-aging effects of P. emblica infusion in a BALB/c mice model. 2) To verify the safety for the consumption of P. emblica infusion in BALB/c mice. MATERIAL AND METHOD: For in vitro studies, splenocytes were isolated from mice and examined in comparison with the human umbilical endothelial cells, fibroblasts and YAC-1 (mouse lymphoma) cells for proliferative activity upon the exposure to P. emblica infusion. For in vivo studies, mice were orally administered with P. emblica infusion at a dose range of 0, 50, 100, 200 mg/kg BW for 14 days. After the treatments, splenocytes isolated from these mice examined for proliferative and NK cell activities. RESULTS: For in vitro studies, the infusion of P. emblica could directly drive the proliferation of mouse splenocytes in a dose-dependent manner. The P. emblica infusion itself was already cytotoxic to YAC-1 in the studied dose, while sparing the human umbilical endothelial cells and fibroblasts. For in vivo studies, splenocytes isolated from these mice exhibited dose-dependent proliferative activities. Only the isolated splenocytes from mice ingesting 100 mg/kg BW exhibited an enhancement in NK cell activity. CONCLUSION: P. emblica infusion could drive proliferative activity of splenocyte in vitro and in vivo, with an enhancement in the NK cell-induced cytotoxic activity. The infusion in the aforementioned dose was safe throughout the study.
Asunto(s)
Citotoxicidad Inmunológica/efectos de los fármacos , Phyllanthus emblica/inmunología , Extractos Vegetales/farmacología , Bazo/citología , Animales , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/inmunología , Masculino , Ratones , Ratones Endogámicos BALB C , Bazo/inmunologíaRESUMEN
BACKGROUND: In vitro polarized human Th2 cells preferentially express the chemokine receptors CCR3 and CCR4 and migrate to their ligands: eotaxin, monocyte-derived chemokine (MDC) or CCL22, thymus- and activation-regulated chemokine (TARC) or CCL17. However little is known about the regulation of these chemokine receptor axes by Der p 1-pulsed dendritic cells in house dust mite (HDM)-sensitive and non-atopic asthmatics. OBJECTIVE: The aim was to investigate the modulatory effects of Der p 1-pulsed DCs on the expression of CCR3 and CCR4 on CD4+ T cells of HDM-sensitive and non-atopic asthmatics. MATERIAL AND METHOD: Using real-time RT-PCR and flow cytometry analysis, the expression of CCR3 and CCR4 were assessed in autologous CD4+ T cells after co-incubation with Der p 1-pulsed DCs from these two asthmatic groups. We also determined the mRNA expression of CCR4 ligands TARC/CCL17 and MDC/CCL22 in monocyte-derived DCs after Der p 1 pulsation. RESULTS: We performed flow cytometry analysis of CD4+ T cells from HDM-sensitive and non-atopic asthmatics, taken 24 and 48 h after co-incubation with Der p 1-pulsed DCs. We demonstrated that after co-incubation, there was a significant increase in CCR3+ and CCR4+ CD4+ T cells from HDM-sensitive asthmatics, which began to occur at 24 h and 48 h respectively, and corresponded to their expression at mRNA levels. In contrast, only CCR4 mRNA but not protein expression was increased in non-atopic CD4+ T cells. After Der p 1 pulsation, mRNA expression of CCR4-specific ligands (CCL17 and CCL22) was also markedly upregulated in HDM-sensitive DCs whereas only CCL17 gene expression was increased in non-atopic DCs. CONCLUSION: These data support the role of DCs in differential regulation of CCR3 and CCR4 on CD4+ T cells from HDM-sensitive and non-atopic asthmatics after Der p 1 exposure.
Asunto(s)
Asma/metabolismo , Células Dendríticas/metabolismo , Receptores CCR3/metabolismo , Receptores CCR4/metabolismo , Alérgenos/inmunología , Alérgenos/metabolismo , Animales , Antígenos Dermatofagoides , Proteínas de Artrópodos , Asma/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Quimiocina CCL17/metabolismo , Quimiocina CCL22/metabolismo , Cisteína Endopeptidasas , Células Dendríticas/inmunología , Citometría de Flujo , Humanos , Pyroglyphidae/genética , Pyroglyphidae/inmunología , Pyroglyphidae/metabolismo , ARN Mensajero , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Th2/inmunología , Células Th2/metabolismoRESUMEN
Cytokine-induced killer (CIK) cells were examined for safety and efficacy for cholangiocarcinoma treatment. Several conditions of human CIK cells were examined using ex vivo cytotoxic assay and SCID mice pre-inoculated with cholangiocarcinoma cells. We monitored the ex vivo cytotoxicity, tumor sizes and immunohistochemistry. Optimal tumor suppression was observed when CIK cells were pre-exposed to dendritic cells (DCs). Unexpectedly, pulsing of tumor RNA to DCs rendered the co-culturing CIK cells ineffective and raised the proportion of CD4(+)CD25(+) subset. The use of CD3(+)CD56(+) subset instead of the whole population of CIK cells for the co-culture with RNA-pulsed DCs restored the efficacy. Tumor-infiltrating human CD3(+) cells were observed from day 2 - 14. The CD3(+)CD56(+) cells are logical candidates for clinical trial while the DC-co-cultured CIK cells produced similar efficacy and more feasible for clinical application. The RNA pulsation of DCs up-regulated the regulatory subset of CIK cells and abrogated the anti-tumor efficacy.
Asunto(s)
Neoplasias de los Conductos Biliares/terapia , Colangiocarcinoma/terapia , Células Asesinas Inducidas por Citocinas/inmunología , Animales , Neoplasias de los Conductos Biliares/patología , Conductos Biliares Intrahepáticos , Complejo CD3/análisis , Antígeno CD56/análisis , Línea Celular Tumoral , Colangiocarcinoma/patología , Células Dendríticas/inmunología , Humanos , Ratones , Ratones SCID , Trasplante de Neoplasias , Trasplante HeterólogoRESUMEN
BACKGROUND: Indoleamine 2, 3-dioxygenase (IDO), a tryptophan-degrading enzyme in dendritic cells (DCs), mediates an immunosuppressive effect on activated T lymphocytes. However, little is known about the effect of Der p 1 on IDO in human DCs. OBJECTIVE: The aim was to investigate the effect of Der p 1 on the expression and activity of IDO in monocyte-derived DCs from house dust mite (HDM)-sensitive patients with asthma. METHODS: Using real-time RT-PCR and HPLC, the expression and activity of IDO were assessed in TNF-alpha-induced mature DCs from HDM-sensitive and nonatopic patients with asthma in response to Der p 1 exposure ex vivo. We also monitored the alteration of IDO activity in Der p 1-pulsed DCs after the coincubation with autologous T cells. RESULTS: With a reliance on its protease activity, Der p 1 suppressed functional IDO in DCs from HDM-sensitive patients with asthma but enhanced IDO activity in DCs from nonatopic patients with asthma. This suppression was maintained by the reciprocally induced IL-4 from the coculturing autologous HDM-sensitive T cells. Conversely, the upregulation of IDO activity in Der p 1-pulsed DCs was maintained by IFN-gamma released from autologous nonatopic T cells and the regulatory T-cell subset. Der p 1 pulsation to sensitive DCs failed to raise regulatory T cells but raised progenitor fractions from cloned HDM-sensitive CD4(+) cells through direct contact and soluble mediators. CONCLUSION: House dust mite-sensitive DCs exposed to Der p 1 downregulated IDO activity and tipped the T(H)1/T(H)2 cytokine balance toward IL-4, resulting in sustainable IDO suppression.
Asunto(s)
Antígenos Dermatofagoides/inmunología , Asma/inmunología , Células Dendríticas/inmunología , Regulación Enzimológica de la Expresión Génica/inmunología , Indolamina-Pirrol 2,3,-Dioxigenasa/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Antígenos Dermatofagoides/farmacología , Proteínas de Artrópodos , Asma/enzimología , Células Cultivadas , Cisteína Endopeptidasas , Células Dendríticas/enzimología , Femenino , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Humanos , Indolamina-Pirrol 2,3,-Dioxigenasa/biosíntesis , Interferón gamma/inmunología , Interleucina-4/inmunología , Masculino , Linfocitos T Colaboradores-Inductores/enzimología , Factor de Necrosis Tumoral alfa/inmunología , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/inmunologíaRESUMEN
Osteosarcoma with distant metastases at late stage has posed a challenge for novel therapeutic modalities. The application of cytokine-induced killer (CIK) cells to osteosarcoma constitutes a promising strategy. This approach had been studied in multiple myeloma and breast cancers, where CIK cells exhibited specific cytotoxicity toward malignant cells while sparing wild-type tissues. However, the consistency of CIK cell-induced anti-tumor cytotoxicity has not been thoroughly examined. We investigated whether autologous CIK cells could effectively induce cytolysis of cultured osteosarcoma cells. In addition to the observed CIK cell-induced osteosarcoma cytolysis, the pre-incubation of CIK cells with autologous dendritic cells pulsed with tumor's total RNA further enhanced the tumor cytolysis to greater than 6-fold. The anti-tumor cytolysis was optimized in complete autologous setting, and was attenuated with allogeneic components. The advantage of the co-culture with RNA-pulsed DC was lost when high CIK cell density was employed for anti-tumor cytotoxic assay, but was maintained in purified CD3(+)CD56(+) cells isolated from the CIK cells. This finding implied that CIK cells at limited cell density could induce effective osteosarcoma cytolysis with an aid from tumor antigen presentation on dendritic cell surface.