RESUMEN

Cancer stem cells (CSCs) play an essential role in cancer development, metastasis, relapse, and resistance to treatment. In this article, the effects of three synthesized ZnO nanofluids on proliferation, apoptosis, and stemness markers of breast cancer stem-like cells are reported. The antiproliferative and apoptotic properties of ZnO nanoparticles were evaluated on breast cancer stem-like cell-enriched mammospheres by MTS assay and flowcytometry, respectively. The expression of stemness markers, including WNT1, NOTCH1, ß-catenin, CXCR4, SOX2, and ALDH3A1 was assessed by real-time PCR. Western blotting was used to analyze the phosphorylation of Janus kinase 2 (JAK2) and Signal Transducer and Activator of Transcription 3 (STAT3). Markers of stemness were significantly decreased by ZnO nanofluids, especially sample (c) with code ZnO-148 with a different order of addition of polyethylene glycol solution at the end of formulation, which considerably decreased all the markers compared to the controls. All the studied ZnO nanofluids considerably reduced viability and induced apoptosis of spheroidal and parental cells, with ZnO-148 presenting the most effective activity. Using CD95L as a death ligand and ZB4 as an extrinsic apoptotic pathway blocker, it was revealed that none of the nanoparticles induced apoptosis through the extrinsic pathway. Results also showed a marked inhibition of the JAK/STAT pathway by ZnO nanoparticles; confirmed by downregulation of Mcl-1 and Bcl-XL expression. The present data demonstrated that ZnO nanofluids could combat breast CSCs via decreasing stemness markers, stimulating apoptosis, and suppressing JAK/STAT activity.

Asunto(s)

Antineoplásicos/farmacología , Neoplasias de la Mama/patología , Nanopartículas , Células Madre Neoplásicas/efectos de los fármacos , Puntos Cuánticos , Óxido de Zinc/farmacología , Antineoplásicos/administración & dosificación , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Portadores de Fármacos , Proteína Ligando Fas/efectos de los fármacos , Humanos , Péptidos y Proteínas de Señalización Intracelular/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Óxido de Zinc/administración & dosificación

RESUMEN

Previously we have shown inhibition of endometrial cancer cell growth with progesterone and calcitriol. However, the mechanisms by which the two agents attenuate proliferation have not been well characterized yet. Herein, we investigated how progesterone and calcitriol induce apoptosis in cancer cells. DNA fragmentation was upregulated by progesterone and calcitriol in ovarian and endometrial cancer cells. Time-dependent treatment of ovarian cancer cells, ES-2, and TOV-21G with progesterone enhanced caspase -8 activity after 12 h, whereas OV-90, TOV-112D, HEC-1A, and HEC-59 cells showed increased activity after 24 h. Caspase 9 activity was increased in all cell lines after 24 h treatment with calcitriol. Pretreatment of cancer cells with a caspase-8 inhibitor (z-IETD-fmk) or caspase-9 inhibitor (Z-LEHD-fmk) significantly attenuated progesterone and calcitriol induced caspase-8 and caspase-9 expression, respectively. The expression of FasL, Fas, FAD, and pro-caspase-8, which constitute the death-inducing signaling complex (DISC), was upregulated in progesterone treated cancer cells. Knockdown of FAS or FADD with specific siRNAs significantly blocked progesterone-induced caspase-8. Cleavage of the BID was not affected by caspase-8 activation suggesting the absence of cross-talk between caspase-8 and caspase-9 pathways. Calcitriol treatment decreased mitochondrial membrane potential and increased the release of cancer cytochrome C. These findings indicate that progesterone induces apoptosis through activation of caspase-8 and calcitriol through caspase-9 activation in cancer cells. A combination of progesterone-calcitriol activates both extrinsic and intrinsic apoptotic pathways in cancer cells.

Asunto(s)

Apoptosis/efectos de los fármacos , Caspasas , Neoplasias Endometriales/metabolismo , Neoplasias Ováricas/metabolismo , Progesterona/farmacología , Calcitriol/metabolismo , Caspasa 8/efectos de los fármacos , Caspasa 8/metabolismo , Caspasa 9/efectos de los fármacos , Caspasa 9/metabolismo , Caspasas/efectos de los fármacos , Caspasas/metabolismo , Línea Celular Tumoral , Citocromos c/efectos de los fármacos , Citocromos c/metabolismo , Proteínas Adaptadoras de Señalización del Receptor del Dominio de Muerte/efectos de los fármacos , Proteínas Adaptadoras de Señalización del Receptor del Dominio de Muerte/metabolismo , Superfamilia de los Dominios de Muerte/efectos de los fármacos , Neoplasias Endometriales/tratamiento farmacológico , Proteína Ligando Fas/efectos de los fármacos , Proteína Ligando Fas/metabolismo , Femenino , Humanos , Técnicas In Vitro , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Neoplasias Ováricas/tratamiento farmacológico , Transducción de Señal/efectos de los fármacos , Receptor fas/efectos de los fármacos , Receptor fas/metabolismo

RESUMEN

Early hepatocyte death occurs in most liver injury cases and triggers liver inflammation, which in combination with other risk factors leads to the development of liver disease. However, the pathogenesis of early phase hepatocyte death remains poorly understood. Here, C57BL/6J mice were treated with the hepatotoxic drug flucloxacillin (FLUX) and the toll-like receptor 9 agonist CpG oligodeoxynucleotide (ODN) to reproduce the early phase of drug-induced hepatotoxicity and investigate its pathogenesis. C57BL/6J mice were treated with FLUX (100 mg/kg, gavage) alone or in combination with ODN (40 µg/mouse, intraperitoneally). Plasma alanine aminotransferase (ALT) level was measured as a marker of hepatotoxicity. FLUX or ODN alone was insufficient to induce ALT elevation, whereas combination treatment with FLUX and ODN increased ALT levels 24 h after FLUX treatment and upregulated Fas ligand in natural killer T (NKT) cells and Fas in hepatocytes. FLUX induced mitochondrial permeability transition (MPT), and pretreatment with ODN sensitized mitochondria to FLUX-induced MPT. The increase in ALT levels induced by ODN and FLUX co-treatment was suppressed in Fas ligand (gld/gld)-deficient mice and in mice deficient in a component of MPT pore opening (cyclophilin D-knockout mice). These results suggested that ODN activated the Fas/Fas ligand-mediated pathway in NKT cells and hepatocytes, which may predispose to FLUX-induced mitochondrial dysfunction and lead to early phase hepatocyte apoptosis. Taken together, these findings elucidate a potentially novel mechanism underlying drug-induced early phase hepatocyte death related to the Fas/Fas ligand death receptor pathway and mitochondrial dysfunction.

Asunto(s)

Apoptosis/efectos de los fármacos , Proteína Ligando Fas/efectos de los fármacos , Hígado/efectos de los fármacos , Enfermedades Mitocondriales/patología , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 9/agonistas , Alanina Transaminasa/sangre , Animales , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Proteína Ligando Fas/genética , Femenino , Hepatocitos/efectos de los fármacos , Células Asesinas Naturales/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Enfermedades Mitocondriales/inducido químicamente , Oligodesoxirribonucleótidos

RESUMEN

Hypoxic-ischemic brain damage (HIBD) occurs due to intrauterine hypoxia ischemia influencing the energy supply for fetal brain cells, which affects the metabolism of the brain to make the brain suffer a severe damage. Erythropoietin (EPO), which regulates hemacytopoiesis, is a kind of cytokine. EPO is sensitive to hypoxia ischemia. In this study, we aimed to investigate the effect of EPO on the expression of Fas/FasL in brain tissues of neonatal rats with HIBD. Neonatal rats were assigned randomly to sham, HIBD, and EPO groups. Five time points for observation were 6, 12, 24, 48, and 72 h after the HIBD rat model had been established, respectively. In the HIBD group, Fas/FasL expression began to rise at 6 h, reached the peak at 12-24 h, and dropped from 24 h. In the EPO group, the expression of Fas/FasL was lower than those in HIBD group at 12, 24, and 48 h (P<0.05). Our findings suggest that EPO may reduce cell apoptosis after hypoxic-ischemic damage through reduction of the expression of Fas and FasL, and that optimal therapeutic time window is 6-24 h after HIBD.

Asunto(s)

Eritropoyetina/farmacología , Proteína Ligando Fas/efectos de los fármacos , Hipoxia Fetal/metabolismo , Hipoxia-Isquemia Encefálica/metabolismo , Receptor fas/efectos de los fármacos , Animales , Animales Recién Nacidos , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Proteína Ligando Fas/biosíntesis , Femenino , Masculino , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Receptor fas/biosíntesis

RESUMEN

Betulinic acid (BA) is a naturally occurring pentacyclic lupane group triterpenoid, has reported to protect cerebral ischemia. Present study evaluates the protective effect of betulinic acid on the isoflurane-induced neuronal damage in neonatal mice. All the mice of 7days age were exposed to isoflurane (2%; 2h) for the duration of 3day. At the end of protocol cognitive function was evaluated by Morris water maze (MWM) test. However count of apoptotic cells were estimated by TUNEL staining. Concentration of oxidative stress parameters [superoxide dismutase (SOD); catalase (CAT) and reduced glutathione (GSH)], cytokines [tumor necrosis factor (TNF-α); interlukin-6 (IL-6) and IL-1ß] and expressions of caspase 3, FAS and FASL were estimated in the neuronal cells. Result of the study suggested that treatment with betulinic acid significantly reduces the escape latency and enhances platform crossing time than negative control group. Count of apoptotic cells were also found to be reduced in BA treated group of mice than negative control group. Moreover treatment with BA significantly attenuates the concentration of inflammatory cytokines and oxidative stress in isoflurane induced neonatal mice. However expressions of caspase-3, FAS/FASL was found to be significantly reduced in BA treated group of mice than negative control group. Present study concludes the neuroprotective effect of betulinic acid in isoflurane-induced brain damage in developing brain by attenuating the apoptosis through Fas/FASL pathway inhibition.

Asunto(s)

Apoptosis/efectos de los fármacos , Proteína Ligando Fas/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Triterpenos/farmacología , Animales , Animales Recién Nacidos , Encéfalo/efectos de los fármacos , Encéfalo/patología , Citocinas/metabolismo , Femenino , Etiquetado Corte-Fin in Situ , Isoflurano/toxicidad , Aprendizaje por Laberinto/efectos de los fármacos , Ratones , Neuronas/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Triterpenos Pentacíclicos , Ratas , Transducción de Señal/efectos de los fármacos , Ácido Betulínico

RESUMEN

OBJECTIVE: To observe the effects of Qiangjing Tablets (QJT) on the semen quality and Fas/FasL signaling pathway in male SD rats with infertility. METHODS: Models of infertility were made in 50 male SD rats by intragastric administration of Tripterygium (GTW) for 3 weeks, and another 20 rats were taken as blank controls. Then 40 successfully established rat models were randomly divided into four groups, model control, low-dose QJT, medium-dose QJT, and high-dose QJT, the latter three groups treated intragastrically with QJT at 58 mg, 105 mg, and 233 mg per kg of the body weight per day, respectively. After 4 weeks of medication, the rats were killed for examination of semen quality and determination of the expression of the apoptosis factor FasL in the testis tissue. RESULTS: Compared with the blank controls, the model rats showed significant decreases in sperm concentration ([71.99 ± 9.72] vs [10.94 ± 3.58] x 106/ml, P < 0.01), motility ([48.95 ± 4.10] vs [9.31 ± 5.79]%, P < 0.01), and viability ( [82.06 ± 6.16] vs [24.03 ± 6.93]%, P < 0.01). In comparison with the model controls, the rats in the QJT groups exhibited remarkably increased sperm concentration, motility, and viability, more significantly in the high-dose group ([59.66 ± 4.53] x 106/ml, [35.45 ± 5.21] %, and [61.97 ± 9.75]%) and medium-dose group ([40.89 ± 4.90] x 106/ml, [24.41 ± 4.79]%, and [60.06 ± 10.62]%) (P < 0.05 or P < 0.01). The expression of FasL was markedly reduced in the low-, medium-, and high-dose QJT groups (0.5215 ± 0.0189, 0.5371 ± 0.0364, and 0.4556 ± 0.0215) as compared with that of the model controls (0.5989 ± 0.0448 ) (P < 0.05 or P < 0.01). CONCLUSION: By upregulating the Fas/FasL signaling pathway, Tripterygium glycosides may induce the apoptosis of spermatogenic cells and reduce sperm concentration, motility and viability, resulting in infertility. The Chinese medicine Qiangjing Tablets can improve the reproductive function of male rats by decreasing the expression of the apoptosis factor FasL in the testis.

Asunto(s)

Medicamentos Herbarios Chinos/farmacología , Proteína Ligando Fas/efectos de los fármacos , Infertilidad Masculina/tratamiento farmacológico , Semen/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Proteína Ligando Fas/metabolismo , Células Germinativas , Glicósidos , Infertilidad Masculina/inducido químicamente , Infertilidad Masculina/metabolismo , Masculino , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Análisis de Semen , Transducción de Señal , Recuento de Espermatozoides , Motilidad Espermática/efectos de los fármacos , Comprimidos , Testículo/efectos de los fármacos , Testículo/metabolismo , Tripterygium

RESUMEN

BACKGROUND: Breastfeeding is associated with a variety of positive health outcomes in children and is recommended exclusively for the first 6 months of life; however, 50-70 % of infants in the US are formula-fed. To test the hypothesis that immune system development and function in neonates and infants are significantly influenced by diet, 2-day old piglets were fed soy or milk formula (n = 6/group/gender) until day 21 and compared to a sow-fed group (n = 6/gender). METHODS: Histomorphometric analyses of ileum, jejunum and Peyer's patches were carried out, to determine the inflammation status, mRNA and protein expression of pro-inflammatory, anti-inflammatory and growth-related chemokines and cytokines. RESULTS: In formula-fed animals, increases in ileum and jejunum villus height and crypt depth were observed in comparison to sow-fed animals (jejunum, p < 0.01 villus height, p < 0.04 crypt depth; ileum p < 0.001 villus height, p < 0.002 crypt depth). In formula-fed the lymphoid follicle size (p < 0.01) and germinal centers (p < 0.01) with in the Peyer's patch were significantly decreased in comparison to sow-fed, indicating less immune education. In ileum, formula diet induced significant up-regulation of AMCFII, IL-8, IL-15, VEGFA, LIF, FASL, CXCL11, CCL4, CCL25 and down-regulation of IL-6, IL-9, IL-10, IL-27, IFNA4, CSF3, LOC100152038, and LOC100736831 at the transcript level. We have confirmed some of the mRNA data by measuring protein, and significant down-regulation of anti-inflammatory molecule IL-10 in comparison to sow-fed piglets was observed. To further determine the membrane protein expression in the ileum, VE-cadherin, occludin, and claudin-3, Western blot analyses were conducted. Sow fed piglets showed significantly more VE-Cadherin, which associated with levels of calcium, and putrescine measured. It is possible that differences in GI tract and immune development are related to shifts in the microbiome; notably, there were 5-fold higher amounts of Lactobacillaceae spp and 3 fold higher Clostridia spp in the sow fed group in comparison to milk formula-fed piglets, whereas in milk formula-fed pigs Enterobacteriaceae spp was 5-fold higher. CONCLUSION: In conclusion, formula diet alters GI morphology, microbial abundance, intestinal barrier protein VE-cadherin and anti-inflammatory molecule IL-10 expression. Further characterization of formula effects could lead to modification of infant formula to improve immune function, reduce inflammation and prevent conditions such as allergies and infections.

Asunto(s)

Antígenos CD/genética , Cadherinas/genética , Citocinas/efectos de los fármacos , Microbioma Gastrointestinal/efectos de los fármacos , Fórmulas Infantiles/farmacología , Intestino Delgado/efectos de los fármacos , Leche , ARN Mensajero/efectos de los fármacos , Alimentos de Soja , Animales , Animales Recién Nacidos , Antígenos CD/metabolismo , Cadherinas/metabolismo , Calcio/metabolismo , Citocinas/genética , Citocinas/metabolismo , Dieta , Regulación hacia Abajo , Proteína Ligando Fas/efectos de los fármacos , Proteína Ligando Fas/genética , Proteína Ligando Fas/metabolismo , Humanos , Íleon/efectos de los fármacos , Íleon/metabolismo , Íleon/microbiología , Íleon/patología , Recién Nacido , Interferón-alfa/efectos de los fármacos , Interferón-alfa/genética , Interferón-alfa/metabolismo , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-15/genética , Interleucina-15/metabolismo , Interleucina-27/genética , Interleucina-27/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/efectos de los fármacos , Interleucina-8/genética , Interleucina-8/metabolismo , Interleucina-9/genética , Interleucina-9/metabolismo , Intestino Delgado/metabolismo , Intestino Delgado/microbiología , Intestino Delgado/patología , Yeyuno/efectos de los fármacos , Yeyuno/metabolismo , Yeyuno/microbiología , Yeyuno/patología , Factor Inhibidor de Leucemia/efectos de los fármacos , Factor Inhibidor de Leucemia/genética , Factor Inhibidor de Leucemia/metabolismo , Ganglios Linfáticos Agregados/efectos de los fármacos , Ganglios Linfáticos Agregados/inmunología , ARN Mensajero/metabolismo , Porcinos , Regulación hacia Arriba , Factor A de Crecimiento Endotelial Vascular/efectos de los fármacos

RESUMEN

Objective To observe the effect of Longzuan Tongbi Recipe (LTR) on Fas/FasL sys- tems in serum and synovium of collagen-induced arthritis (CIA) rats. Methods Ten rats were randomly selected from 60 male Wistar rats as a normal control group. CIA model was prepared by injecting type II bovine collagen and incomplete Freund's adjuvant mixture in the rest 50 rats. After modeling rats were di- vided into the model group, the methotrexate (MTX) group, high, middle, and low dose LTR groups, 10 in each group. Normal saline was administered to rats in the model group by gastrogavage. MTX solution (0.27 mg/100 g) was administered to rats in the MTX group by gastrogavage, once per week for 4 succes- sive weeks. LTR (4.32, 2.16, 1.08 g/mL) was administered to rats in the 3 LTR groups by gastrogavage, twice per day for 30 successive days. Morphological changes of synovium were observed by HE staining. Expression levels of Fas/FasL in rat serum and synovium were quantitatively detected by ELISA. Results Normal synovium cells could be seen in the normal group. But they were obviously proliferated, fat cells in the lower synovium were reduced or deformed, fibroblasts were increased in the model group, accompa- nied with infiltration of lymphocytes and monocytes. All these changes were more obviously alleviated in the MTX group, and the 3 LTR groups. Compared withI the normal control group, Fas expression level in- creased in rat serum and synovium, serum FasL expression level decreased in the model group (P <0. 05, P <0. 01). Compared with the model group, Fas expression level decreased in rat serum and synovium in the MTX group, high and middle dose LTR groups; Fas expression level in rat serum increased in the MTX group and 3 LTR groups; Fas expression level in synovium increased in the MTX group, high and middle dose LTR groups (P <0. 05, P <0. 01). Compared with the MTX group, Fas expression level in serum of the low dose LTR group, and Fas expression level in synovium of low and middle dose LTR groups was elevat- ed; Fas expression level in serum and synovium of the high dose LTR group was reduced; FasL expres- sion level in serum and synovium of low and middle dose LTR groups was reduced; FasL expression level in serum and synovium increased of the high dose LTR group (P <0. 05, P <0. 01). Conclusion LTR could control and treat rheumatoid arthritis, and its mechanism might lie in regulating. Fas/FasL systems media- ted cell apoptosis, and relieving pathological reaction of rheumatoid arthritis.

Asunto(s)

Artritis Experimental , Medicamentos Herbarios Chinos , Membrana Sinovial , Animales , Artritis Experimental/tratamiento farmacológico , Bovinos , Colágeno Tipo II , Medicamentos Herbarios Chinos/farmacología , Proteína Ligando Fas/efectos de los fármacos , Masculino , Distribución Aleatoria , Ratas , Ratas Wistar , Membrana Sinovial/efectos de los fármacos , Receptor fas/efectos de los fármacos

RESUMEN

BACKGROUND: Doxycycline (DC) has been shown to possess non-antibiotic properties including Fas/Fas Ligand (FasL)-mediated apoptosis against several tumor types in the concentration range of 10-40 µg/mL. However, the effect of DC in apoptotic signaling at much low concentrations was not studied. METHODS: The present study investigated the attenuation effect of low dose of DC on FasL-induced apoptosis in HeLa cell by the methods of MTT assay, fluorescence microscopy, DNA fragmentation, flow cytometry analysis, and western blotting. RESULTS AND CONCLUSION: In the present findings we showed that low concentration of DC (<2.0 µg/mL) exhibited protective effects against FasL-induced apoptosis in HeLa cells. FasL treatment to HeLa cells resulted in a concentration-dependent induction of cell death, and treatment with low concentrations of DC (0.1-2 µg/mL) significantly (p < 0.001) attenuated the FasL-induced cell death as measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Further, the FasL-induced apoptotic features in HeLa cells, such as morphological changes, DNA fragmentation and cell cycle arrest was also inhibited by DC (0.5 µg/mL). Tetracycline and minocycline also showed similar anti-apoptotic effects but were not significant when compared to DC, tested at same concentrations. Further, DC (0.01-16 µg/mL) did not influence the hydrogen peroxide- or cisplatin-induced intrinsic apoptotic pathway in HeLa cells. Protein analysis using Western blotting confirmed that FasL-induced cleavage/activation of caspase-8 and caspase-3, were inhibited by DC treatment at low concentration (0.5 µg/mL). Considering the overall data, we report for the first time that DC exhibited anti-apoptotic effects at low concentrations in HeLa cells by inhibition of caspase activation via FasL-induced extrinsic pathway.

Asunto(s)

Apoptosis/efectos de los fármacos , Caspasas/efectos de los fármacos , Doxiciclina/administración & dosificación , Proteína Ligando Fas/efectos de los fármacos , Animales , Western Blotting , Relación Dosis-Respuesta a Droga , Doxiciclina/farmacología , Activación Enzimática , Citometría de Flujo , Células HeLa , Humanos , Ratones , Células 3T3 NIH

RESUMEN

This study investigated the treatment effects of a new compound, strontium fructose 1, 6-diphosphate (FDP-Sr), in cyclophosphamide (CP)-induced oligozoospermia. FDP-Sr, with extra high-energy supply, could reverse male hypogonadism in the testis. Male Wistar rats were randomly divided into three groups: control group (vehicle treated), CP group and CP + FDP-Sr group. Both CP group and CP + FDP-Sr groups were orally administered CP (20 mg kg(-1) ) consecutively for the first 7 days to establish CP-induced testicular toxic models. Subsequently, CP group was given orally distilled water per day, whereas CP + FDP-Sr group was received FDP-Sr (200 mg kg(-1) ) for 49 days. Compared to the CP group, the FDP-Sr group showed significantly increased levels of serum testosterone, testis relative weights and epididymal sperm counts in rats. In addition, rats treated by FDP-Sr showed the recuperative activities of testicular marker enzymes and normalised levels of antioxidants in tissue. Testicular protection of FDP-Sr was further demonstrated by enhancing expression of P450scc, reducing ability of FAS/FASL and generating cytoprotection in the histopathological study. FDP-Sr appeared to possess an ability to attenuate CP-induced reproduction toxicity via the activation of antioxidants and steroidogenesis enzymes, and alleviate oligozoospermia via inhibition of testicular apoptosis by FAS/FASL pathway.

Asunto(s)

Antioxidantes/farmacología , Apoptosis/efectos de los fármacos , Fructosadifosfatos/farmacología , Oligospermia/metabolismo , Testículo/efectos de los fármacos , Animales , Antineoplásicos Alquilantes/toxicidad , Apoptosis/genética , Ciclofosfamida/toxicidad , Modelos Animales de Enfermedad , Proteína Ligando Fas/efectos de los fármacos , Proteína Ligando Fas/genética , Proteína Ligando Fas/metabolismo , Glutatión/efectos de los fármacos , Glutatión/metabolismo , Glutatión Peroxidasa/efectos de los fármacos , Glutatión Peroxidasa/metabolismo , L-Iditol 2-Deshidrogenasa/efectos de los fármacos , L-Iditol 2-Deshidrogenasa/metabolismo , L-Lactato Deshidrogenasa/efectos de los fármacos , L-Lactato Deshidrogenasa/metabolismo , Peróxidos Lipídicos/metabolismo , Masculino , Malondialdehído/metabolismo , Oligospermia/inducido químicamente , Tamaño de los Órganos/efectos de los fármacos , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Recuento de Espermatozoides , Recuperación de la Esperma , Superóxido Dismutasa/efectos de los fármacos , Superóxido Dismutasa/metabolismo , Testículo/anatomía & histología , Testosterona/sangre , Receptor fas/efectos de los fármacos , Receptor fas/genética , Receptor fas/metabolismo , gamma-Glutamiltransferasa/efectos de los fármacos , gamma-Glutamiltransferasa/metabolismo

RESUMEN

Stem cells from exfoliated deciduous teeth (SHED) possess multipotent differentiation and immunomodulatory properties. They have been used for orofacial bone regeneration and autoimmune disease treatment. In this study, we show that acetylsalicylic acid (ASA) treatment is able to significantly improve SHED-mediated osteogenic differentiation and immunomodulation. Mechanistically, ASA treatment upregulates the telomerase reverse transcriptase (TERT)/Wnt/ß-catenin cascade, leading to improvement of SHED-mediated bone regeneration, and also upregulates TERT/FASL signaling, leading to improvement of SHED-mediated T-cell apoptosis and ameliorating disease phenotypes in dextran sodium sulfate-induced colitis mice. These data indicate that ASA treatment is a practical approach to improving SHED-based cell therapy.

Asunto(s)

Aspirina/farmacología , Células Madre Mesenquimatosas/efectos de los fármacos , Diente Primario/citología , Animales , Apoptosis/efectos de los fármacos , Aspirina/administración & dosificación , Regeneración Ósea/efectos de los fármacos , Técnicas de Cultivo de Célula , Diferenciación Celular/efectos de los fármacos , Tratamiento Basado en Trasplante de Células y Tejidos , Niño , Colitis/terapia , Relación Dosis-Respuesta a Droga , Proteína Ligando Fas/efectos de los fármacos , Humanos , Hidroxiapatitas/química , Inmunomodulación/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Ratones Desnudos , Células Madre Multipotentes/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Linfocitos T/efectos de los fármacos , Telomerasa/efectos de los fármacos , Andamios del Tejido/química , Vía de Señalización Wnt/efectos de los fármacos

RESUMEN

BACKGROUND: Doxycycline (DC) has been shown to possess non-antibiotic properties including Fas/Fas Ligand (FasL)-mediated apoptosis against several tumor types in the concentration range of 10-40 µg/mL. However, the effect of DC in apoptotic signaling at much low concentrations was not studied. METHODS: The present study investigated the attenuation effect of low dose of DC on FasL-induced apoptosis in HeLa cell by the methods of MTT assay, fluorescence microscopy, DNA fragmentation, flow cytometry analysis, and western blotting. RESULTS AND CONCLUSION: In the present findings we showed that low concentration of DC (<2.0 µg/mL) exhibited protective effects against FasL-induced apoptosis in HeLa cells. FasL treatment to HeLa cells resulted in a concentration-dependent induction of cell death, and treatment with low concentrations of DC (0.1-2 µg/mL) significantly (p < 0.001) attenuated the FasL-induced cell death as measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazo-lium bromide (MTT) assay. Further, the FasL-induced apoptotic features in HeLa cells, such as morphological changes, DNA fragmentation and cell cycle arrest was also inhibited by DC (0.5 µg/mL). Tetracycline and minocycline also showed similar anti-apoptotic effects but were not significant when compared to DC, tested at same concentrations. Further, DC (0.01-16 µg/mL) did not influence the hydrogen peroxide- or cisplatin-induced intrinsic apoptotic pathway in HeLa cells. Protein analysis using Western blotting confirmed that FasL-induced cleavage/activation of cas-pase-8 and caspase-3, were inhibited by DC treatment at low concentration (0.5 µg/mL). Considering the overall data, we report for the first time that DC exhibited anti-apoptotic effects at low concentrations in HeLa cells by inhibition of caspase activation via FasL-induced extrinsic pathway.

Asunto(s)

Humanos , Animales , Ratones , Apoptosis/efectos de los fármacos , Doxiciclina/administración & dosificación , Caspasas/efectos de los fármacos , Proteína Ligando Fas/efectos de los fármacos , Células HeLa , Western Blotting , Doxiciclina/farmacología , Células 3T3 NIH , Relación Dosis-Respuesta a Droga , Activación Enzimática , Citometría de Flujo

RESUMEN

Alkaloids are the most extensively featured compounds of natural anti-tumor herbs, which have attracted much attention in pharmaceutical research. In our previous studies, a mixture of major three alkaloid components (5, 6-dihydrobicolorine, 7-deoxy-trans-dihydronarciclasine, littoraline) from Hymenocallis littoralis were extracted, analyzed and designated as AHL. In this paper, AHL extracts were added to human liver hepatocellular cells HepG-2, human gastric cancer cell SGC-7901, human breast adenocarcinoma cell MCF-7 and human umbilical vein endothelial cell EVC-304, to screen one or more AHL-sensitive tumor cell. Among these cells, HepG-2 was the most sensitive to AHL treatment, a very low dose (0.8µg/ml) significantly inhibiting proliferation . The non- tumor cell EVC-304, however, was not apparently affected. Effect of AHL on HepG-2 cells was then explored. We found that the AHL could cause HepG-2 cycle arrest at G2/M checkpoint, induce apoptosis, and interrupt polymerization of microtubules. In addition, expression of two cell cycle-regulated proteins, CyclinB1 and CDK1, was up-regulated upon AHL treatment. Up-regulation of the Fas, Fas ligand, Caspase-8 and Caspase-3 was observed as well, which might imply roles for the Fas/FsaL signaling pathway in the AHL-induced apoptosis of HepG-2 cells.

Asunto(s)

Alcaloides/farmacología , Apoptosis/efectos de los fármacos , Proteína Ligando Fas/efectos de los fármacos , Liliaceae , Transducción de Señal/efectos de los fármacos , Apoptosis/genética , Western Blotting , Proliferación Celular/efectos de los fármacos , Supervivencia Celular , Proteína Ligando Fas/genética , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Células Hep G2/citología , Células Hep G2/efectos de los fármacos , Humanos , Extractos Vegetales , Valores de Referencia , Sensibilidad y Especificidad , Transducción de Señal/genética , Células Tumorales Cultivadas

RESUMEN

OBJECTIVE: Injection of agonistic anti-Fas antibody has been shown to decrease disease symptoms in mouse models of arthritis. Additionally, membrane-bound FasL (mFasL) has been shown to induce cell death in fibroblast-like synoviocytes (FLS) from rheumatoid arthritis (RA) patients. However, levels of soluble FasL (sFasL) are increased in the joints of RA patients and have been associated with disease severity, indicating that mFasL and sFasL play opposing roles in RA. The purpose of this study was to analyze the effects of FasL on RA FLS responses. METHODS: The responses of FLS from RA and osteoarthritis (OA) patients to soluble and oligomeric FasL, the latter mimicking mFasL, were analyzed by fluorescence-activated cell sorting and proliferation assays, using 3 different FasL variants. The signaling pathways that trigger FasL responses were characterized by Western blotting. RESULTS: We found that mFasL and sFasL have distinct roles in RA FLS. Crosslinked FasL preferentially induced apoptosis, whereas sFasL stimulated proliferation. Moreover, sFasL activated several signaling pathways in RA FLS, such as ERK-1/2, phosphatidylinositol 3-kinase, caspase 8, and JNK, with a prominent role of JNK, since only the blockade of this pathway rendered FLS more susceptible to FasL-induced apoptosis. Crosslinked FasL induced apoptosis in FLS from OA patients, but sFasL failed to stimulate their proliferation. CONCLUSION: Our findings suggest that sFasL is a disease promoter in RA, a finding consistent with previous reports describing a tumor-promoting role of FasL. Therefore, blocking of sFasL could be a therapeutic strategy for RA.

Asunto(s)

Apoptosis/fisiología , Artritis Reumatoide/metabolismo , Proliferación Celular/fisiología , Proteína Ligando Fas/metabolismo , Fibroblastos/metabolismo , Membrana Sinovial/metabolismo , Adulto , Anciano , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Proteína Ligando Fas/efectos de los fármacos , Femenino , Fibroblastos/efectos de los fármacos , Citometría de Flujo , Humanos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/fisiología , Masculino , Persona de Mediana Edad , Osteoartritis/metabolismo , Fosfatidilinositol 3-Quinasas/efectos de los fármacos , Fosfatidilinositol 3-Quinasas/fisiología , Membrana Sinovial/citología , Membrana Sinovial/efectos de los fármacos

RESUMEN

BACKGROUND: Glutamate excitotoxicity and cyclic AMP-activated protein kinase (AMPK) are both recognized as important mediators in neurodegenerative disorders including Alzheimer's disease (AD). OBJECTIVES: To investigate whether oral or subcutaneous monosodium glutamate (MSG) neurotoxicity mimics some features of AD and whether these can be reversed by the AMPK activator Pioglitazone. METHODS: Male Wistar rats aged 5 weeks were administered oral or subcutaneous MSG for 10 days with or without daily oral Pioglitazone. Two additional groups given only saline orally or subcutaneously acted as controls. At age 10 weeks the rats were subjected to neurobehavioral testing, then sacrificed for measurement of AMPK, ß-amyloid and Fas ligand in the hippocampus. RESULTS: Oral and subcutaneous MSG both induced a lowering of hippocampal AMPK by 43% and 31% respectively (P<0.05 for both) and >2-fold increase in hippocampal Fas ligand, a mediator of apoptosis (P<0.001 for both). MSG treatment also induced a significant increase in ß-amyloid in the hippocampus by >4-fold and >5-fold in the oral and subcutaneous groups. This was associated with increased latency before crossing to the white half in the black-white alley and before the first rear in the holeboard test, suggesting increased anxiety. Pioglitazone decreased hippocampal ß-amyloid accumulation and Fas ligand, but did not ameliorate the neurobehavioural deficits induced by MSG. CONCLUSIONS: MSG treatment enhances ß-amyloid accumulation in the rat hippocampus. Our results suggest a role for AMPK reduction in mediating the neurotoxic effects of glutamate, including ß-amyloid accumulation.

Asunto(s)

Proteínas Quinasas Activadas por AMP/metabolismo , Péptidos beta-Amiloides/metabolismo , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Degeneración Nerviosa/inducido químicamente , Glutamato de Sodio/toxicidad , Animales , Conducta Animal/efectos de los fármacos , Proteína Ligando Fas/efectos de los fármacos , Proteína Ligando Fas/metabolismo , Ácido Glutámico/metabolismo , Masculino , Degeneración Nerviosa/metabolismo , Pioglitazona , Ratas , Glutamato de Sodio/antagonistas & inhibidores , Tiazolidinedionas/farmacología

RESUMEN

BACKGROUND: ß-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ß-elemene against glioma cells remains unclear. In the present study, we assessed effects of ß-elemene on human glioma cells and explored the underlying mechanism. MATERIALS AND METHODS: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ß-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ß-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ß-elemne treatment group. RESULTS: With increase in the concentration of ß-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability (IC50) was 48.5 µg/mL for 24h. ß-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ß-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ß-elemene in a time and does- dependent manner. CONCLUSIONS: Our results indicate that ß-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

Asunto(s)

Apoptosis/efectos de los fármacos , Proteína Ligando Fas/efectos de los fármacos , Glioma/genética , Proteínas Proto-Oncogénicas c-bcl-2/efectos de los fármacos , ARN Mensajero/efectos de los fármacos , Sesquiterpenos/farmacología , Proteína X Asociada a bcl-2/efectos de los fármacos , Receptor fas/efectos de los fármacos , Apoptosis/genética , Western Blotting , Caspasas/efectos de los fármacos , Caspasas/genética , Caspasas/metabolismo , Línea Celular Tumoral , Regulación hacia Abajo/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Proteína Ligando Fas/genética , Proteína Ligando Fas/metabolismo , Glioma/metabolismo , Humanos , Técnicas In Vitro , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Regulación hacia Arriba/efectos de los fármacos , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismo , Receptor fas/genética , Receptor fas/metabolismo

RESUMEN

Lenalidomide is a potent immunomodulatory agent capable of downregulating proinflammatory cytokines such as tumor necrosis factor-α (TNF-α) and upregulating anti-inflammatory cytokines. Lenalidomide has been shown to elicit cardiovascular effects, although its impact on cardiac function remains obscure. This study was designed to examine the effect of lenalidomide on cardiac contractile function in ob/ob obese mice. C57BL lean and ob/ob obese mice were given lenalidomide (50 mg/kg/day, p.o.) for 3 days. Body fat composition was assessed by dual-energy X-ray absorptiometry. Cardiomyocyte contractile and intracellular Ca(2+) properties were evaluated. Expression of TNF-α, interleukin-6 (IL-6), Fas, Fas ligand (FasL), the short-chain fatty acid receptor GPR41, the NFκB regulator IκB, endoplasmic reticulum (ER) stress, the apoptotic protein markers Bax, Bcl-2, caspase-8, tBid, cytosolic cytochrome C, and caspase-12; and the stress signaling molecules p38 and extracellular signal-regulated kinase (ERK) were evaluated by western blot. ob/ob mice displayed elevated serum TNF-α and IL-6 levels, fat composition and glucose intolerance, the effects of which except glucose intolerance and fat composition were attenuated by lenalidomide. Cardiomyocytes from ob/ob mice exhibited depressed peak shortening (PS) and maximal velocity of shortening/relengthening, prolonged time-to-PS and time-to-90% relengthening as well as intracellular Ca(2+) mishandling, which were ablated by lenalidomide. Western blot analysis revealed elevated levels of TNF-α, IL-6, Fas, Bip, Bax, caspase-8, tBid, cleaved caspase-3 caspase-12, cytochrome C, phosphorylation of p38, and ERK in ob/ob mouse hearts, the effects of which with the exception of Bip, Bax, and caspase-12 were alleviated by lenalidomide. Taken together, these data suggest that lenalidomide is protective against obesity-induced cardiomyopathy possibly through antagonism of cytokine/Fas-induced activation of stress signaling and apoptosis.

Asunto(s)

Caspasas Efectoras/efectos de los fármacos , Contracción Miocárdica/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Obesidad/tratamiento farmacológico , Talidomida/análogos & derivados , Factor de Necrosis Tumoral alfa/efectos de los fármacos , Absorciometría de Fotón , Animales , Western Blotting , Células Cultivadas , Retículo Endoplásmico/efectos de los fármacos , Proteína Ligando Fas/efectos de los fármacos , Interleucina-6/metabolismo , Lenalidomida , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Miocitos Cardíacos/metabolismo , Obesidad/metabolismo , Obesidad/fisiopatología , Transducción de Señal , Talidomida/farmacología

RESUMEN

PURPOSE: We investigated the function of Fas in photoreceptors. METHODS: Postmortem human eyes and mouse-derived photoreceptor cells (661W) were examined for Fas expression by in situ hybridization and immunofluorescence. 661W cells were treated with FasL or Fas agonistic antibody, or exposed to light with/without pharmacological manipulation of Fas signaling, followed by apoptosis detection by TUNEL, immunofluorescence and fluorescence activated cell scanning (FACS). Fractionated cellular extracts were used to detect protein expression or protein phosphorylation after immunoprecipitation by Western blot. RESULTS: Fas was expressed in the photoreceptor layer of human retina. Fas and a cleaved form of FasL were found on the cell surface of 661W cells. Treatment with FasL or Fas agonistic antibody induced apoptosis in 661W cells. Blocking the activity of FasL or administration of caspase-8 inhibitor z-IETD inhibited light-induced apoptosis. However, it simultaneously caused induction of necroptosis, which could be blocked by the receptor-interacting protein 1 (RIP1) inhibitor, necrostatin-1. Light exposure in the presence of z-IETD caused hyper-phosphorylation of RIP1. Light exposure did not elevate the expression of Fas, FasL, or the Fas-associated death domain adaptor protein (FADD). Cells or conditioned medium after light exposure induced apoptosis in dark-adapted cells, which could be attenuated by blockade of Fas. CONCLUSIONS: Fas has a pro-apoptotic role in photoreceptors. Under light stress, soluble and membrane-bound FasL can bind to Fas, inducing apoptosis via a paracrine mechanism. Although blocking Fas signaling inhibits apoptosis, it does not improve the overall photoreceptor survival due to a compensatory activation of necroptosis. Hence, prevention of photoreceptor loss from retinal photo-oxidative stress should target Fas and RIP1.

Asunto(s)

Apoptosis/fisiología , Proteína Ligando Fas/metabolismo , Luz/efectos adversos , Células Fotorreceptoras/efectos de los fármacos , Receptor fas/fisiología , Animales , Apoptosis/efectos de los fármacos , Western Blotting , Inhibidores de Caspasas , Inhibidores de Cisteína Proteinasa/farmacología , Proteína Ligando Fas/efectos de los fármacos , Humanos , Imidazoles/farmacología , Inmunoprecipitación , Etiquetado Corte-Fin in Situ , Indoles/farmacología , Ratones

RESUMEN

BACKGROUND: CD4 gains in HIV patients on HAART result from release of T cells recently migrated from the thymus, redistribution from lymphoid tissues, proliferation in the periphery and/or reduced apoptosis. The relative contribution of each mechanism in CD4 restoration in patients with suppressed viremia switching antiretrovirals is unclear. METHODS: HIV patients with undetectable viremia on HAART were identified at our clinic. A subset switched to raltegravir was compared with another group that kept therapy unmodified. Naive and memory CD4 T-cells were measured by flow cytometry using CD45RA and CD27, respectively. Activation was examined using CD38 and recent thymic emigrants using CD31. Apoptosis was analyzed measuring soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Fas ligand (FasL). RESULTS: Thirty-seven patients were examined, 19 switched to raltegravir and 18 controls, after a median of 26 months of suppressed viremia. At 6 months, mean CD4 cell counts significantly increased in raltegravir patients from 322 to 448 cells/µl (P = 0.026) but not in controls (from 312 to 330 cells/µl; P  = 0.813). No significant changes were recognized in activation or CD31 expression in any group. In raltegravir patients, however, the proportion of naive CD4 T cells significantly increased (P = 0.014) as well as CD38 expression in these cells (P = 0.036). A positive correlation was found between CD38 and CD31 expression in naive CD4 T cells (R  = 0.51, P < 0.001). TRAIL and FasL did not decline significantly in any group. CONCLUSION: HIV patients with prolonged undetectable viremia on HAART experience more pronounced CD4 gains after raltegravir switching than keeping the same regimen. An increased production of naive CD4 T cells largely explains this effect.

Asunto(s)

Infecciones por VIH/inmunología , Inhibidores de Integrasa VIH/uso terapéutico , Pirrolidinonas/uso terapéutico , ADP-Ribosil Ciclasa 1/efectos de los fármacos , ADP-Ribosil Ciclasa 1/inmunología , Adulto , Terapia Antirretroviral Altamente Activa/métodos , Recuento de Linfocito CD4 , Estudios de Casos y Controles , Proteína Ligando Fas/efectos de los fármacos , Proteína Ligando Fas/inmunología , Citometría de Flujo , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , Humanos , Antígenos Comunes de Leucocito/efectos de los fármacos , Antígenos Comunes de Leucocito/inmunología , Persona de Mediana Edad , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/efectos de los fármacos , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/inmunología , Raltegravir Potásico , Ligando Inductor de Apoptosis Relacionado con TNF/efectos de los fármacos , Ligando Inductor de Apoptosis Relacionado con TNF/inmunología , Resultado del Tratamiento , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/efectos de los fármacos , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/inmunología

RESUMEN

Features of airway remodelling have been described using tissue obtained from fatal cases of asthma and bronchial biopsies from mildly, moderately and severely asthmatic patients. Epithelial detachment and smooth muscle mass enhancement are common features of asthmatic bronchial tissue. This study is the first to investigate the inhibitory effect of nonylphenol (NP) on human bronchial epithelial cell lines BEAS-2B and HBE135-E6E7 (HBE). The results show that NP inhibits bronchial epithelial proliferation via the Fas/Fas ligand apoptotic system. We also treated BEAS-2B and HBE with NP and harvested the condition medium (CM), which was then added to bronchial smooth muscle cells (BSMC). Cultures of BSMC with NP-BEAS-2B-CM and NP-HBE-CM increased BSMC proliferation and migration. Exposure of BEAS-2B and HBE to NP caused epithelial cells to produce inflammatory cytokines IL-6 and IL-8, which subsequently induced BSMC proliferation and migration. Depleting both IL-6 and IL-8 completely reversed the effect of NP-BEAS-2B-CM- and NP-HBE-CM-mediated BSMC proliferation and migration, suggesting that this effect is a synergistic influence of IL-6 and IL-8. This study is the first to demonstrate that NP not only induces bronchial epithelial apoptosis via the Fas-mediated pathway but also stimulates the bronchial epithelium to secrete IL-6 and IL-8, which cause bronchial smooth muscle proliferation and migration - major features in asthma remodelling.

Asunto(s)

Apoptosis/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Fenoles/farmacología , Mucosa Respiratoria/efectos de los fármacos , Caspasa 8/genética , Caspasa 8/metabolismo , Línea Celular , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Proteína Ligando Fas/efectos de los fármacos , Proteína Ligando Fas/metabolismo , Humanos , Miocitos del Músculo Liso/efectos de los fármacos , Miocitos del Músculo Liso/metabolismo , Mucosa Respiratoria/metabolismo