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1.
Anal Chim Acta ; 1294: 342293, 2024 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-38336414

RESUMEN

BACKGROUND: The practical application of moisture sensitive metal organic frameworks (MOFs) in extraction technology faces challenges related to competitive adsorption and water stability. The target analytes cannot be effectively extracted under humid conditions due to the competitive moisture adsorption and/or framework structure collapse of MOFs. In this study, the microporous organic networks (MONs) were synthesized through Sonogashira coupling reaction to use for hydrophobic modification on the surface of MOF-199. RESULTS: The MOF-199@MON as coating was deposited on stainless steel wires for solid-phase microextraction (SPME) of benzene series (BTEX) in aqueous environments. Under the optimal extraction conditions, the MOF-199@MON coated fiber for SPME coupled with GC-MS for the determination of BTEX gave the linear range of 0.5-500 µg L-1, the limit of detections (LODs, S/N = 3) of 0.01-0.04 µg L-1, the limit of quantifications (LOQs, S/N = 10) of 0.04-0.12 µg L-1, the enhancement factors of 3567-4878, and the intra-day, inter-day and fiber-to-fiber precisions (relative standard deviations, RSDs) of 1.0-9.8, 1.9-7.9 and 4.5-9.5 %, respectively. The developed method was successfully applied to the analysis of BTEX in water samples with the recoveries of 71.0 %-113 %. SIGNIFICANCE: This work reveals the home-made SPME fibers have a long service life (the extraction efficiency of fiber decreased by only 7.26 %-13.14 % after 100 cycles). The potential of MON functionalized MOFs as effective adsorbents for the SPME of pollutants in the water environment.

2.
Exp Biol Med (Maywood) ; 248(3): 201-208, 2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-36598065

RESUMEN

This study set out to investigate the clinical significance of serum tumor necrosis factor receptor-associated protein 1 (TRAP1) in diagnosing small cell lung cancer (SCLC) with different clinical stages, and to compare the diagnostic efficiency with neuron-specific enolase (NSE), carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9). Besides, to analyze the role of serum TRAP1 in tumor immunity. A total of 91 patients with SCLC, 99 patients with non-small cell lung cancer (NSCLC), 102 patients with pulmonary nodules (PN), and 75 healthy people were included. The concentrations of serum TRAP1 was detected by enzyme-linked immunosorbent assay (ELISA). NSE, CEA, and CA19-9 were detected by chemiluminescence. The results showed that level of TRAP1 in Group SCLC was lower than other three groups (P < 0.01), whereas NSE in SCLC was significantly higher than the others (P < 0.01), and the levels of CEA and CA19-9 were higher than healthy people and PN patients (P < 0.01). There was a significant difference in TRAP1 levels between patients with limited-stage disease SCLC (LD-SCLC) and extensive-stage disease SCLC (ED-SCLC) (P < 0.0001). The sensitivity and specificity of TRAP1 in diagnosing LD-SCLC were 0.964 and 0.560, respectively, and the area under the curve (AUC) was 0.819. The sensitivity and specificity in diagnosing ED-SCLC were 0.810 and 0.868, respectively, and the AUC was 0.933, which showed high diagnostic value. The AUC of these two groups can be increased to 0.946 and 0.947 in combination of four biomarkers, effectively improving the diagnosis rate of SCLC. Our findings have revealed that serum TRAP1 has high diagnostic value for SCLC and high diagnostic sensitivity for LD-SCLC. It is a potential biomarker for SCLC. Combined detection can effectively improve the diagnosis rate of SCLC. TRAP1 may be secreted into the circulation by mature immune cells and participates in tumor immunity as a carrier of tumor antigens.


Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Humanos , Antígeno Carcinoembrionario , Carcinoma de Pulmón de Células no Pequeñas/patología , Neoplasias Pulmonares/patología , Antígeno CA-19-9 , Biomarcadores de Tumor/análisis , Proteínas HSP90 de Choque Térmico
3.
ACS Appl Mater Interfaces ; 15(4): 5990-5997, 2023 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-36689469

RESUMEN

Here, two-dimensional (2D) nitrogen-doped carbon nanosheets (CNSs) were prepared through carbonizing MOFs (ZIF-8) in-situ grown using graphitic carbon nitride (g-C3N4) as a template. The developed ZIF-8 CNS was then used as solid-phase microextraction (SPME) fiber coating for beneficiation of five biomarkers in exhalation of patients with esophageal cancer and in gas chromatography-mass spectrometry (GC-MS) for determination. The ZIF-8 CNS fiber exhibits satisfactory enrichment factors (3490-5631), wide linearity (5-1000 µg L-1), and low detection limits (0.26-0.96 µg L-1). The relative standard deviations (RSDs) for six replicate extractions of the same ZIF-8 CNS fiber were between 2.0-3.9% (intra-day) and 2.8-5.2% (inter-day). The reproducibility of three fibers prepared by the same approach was in the range 6.8-12.3% (RSD). The developed ZIF-8 CNS fiber can persist in 120 SPME cycles with no prominent loss of extraction efficiency and precision. The high enrichment factors of the 2D ZIF-8 CNS coatings are attributed to the high specific surface area, ultrathin thickness, and nano-pore or interlayer channels; moreover, nitrogen doping also endows the π system with a strong electron absorption ability, which will enhance the π-π interaction between the ZIF-8 CNS and the aromatic ring. Ultimately, the self-made ZIF-8 CNS-coated SPME fiber was applied to the analysis of exhaled breath samples. The recoveries of spiked analytes are between 84 and 105%.


Asunto(s)
Carbono , Neoplasias Esofágicas , Humanos , Carbono/química , Nitrógeno , Microextracción en Fase Sólida/métodos , Espiración , Reproducibilidad de los Resultados
4.
Anal Chim Acta ; 1181: 338901, 2021 Oct 09.
Artículo en Inglés | MEDLINE | ID: mdl-34556209

RESUMEN

In this work, we reported the preparation of the hollow zeolitic imidazolate framework-7 (ZIF-7) via etching ZIF-7 with tannic acid, and further fabricated the hollow ZIF-7 coated fiber for the solid phase microextraction (SPME) of the five volatile biomarkers (acetone, isopropanol, hexanal, hexanol and decanal) generated from breast cancer cell lines. The hollow structure not only endowed higher extraction performance for the SPME of analytes, but also improved the diffusion rate of the analytes inside the hollow ZIF-7. Under the optimal conditions, the hollow ZIF-7 coated fiber offered high extraction capacity (25-153 mg g-1) and enhancement factors (EFs, 2023-11250) for the five biomarkers, good linearity (R2 > 0.9918) of acetone and isopropanol (2.5-500 µg L-1) and hexanol, hexanal, and decanal (1.0-100 µg L-1), low limits of detection (S/N = 3) of 0.07-0.53 µg L-1 and the limit of quantifications (LOQs, S/N = 10) of 0.23-1.76 µg L-1. The precisions (RSDs, %) for intra-day (n = 6), inter-day (n = 5) and fiber-to-fiber (n = 6) were 2.8-7.5%, 4.3-8.5%, and 4.2-14.6%, respectively. The high EFs of the hollow ZIF-7 coated fiber for the five biomarkers resulted from the integrated effects of the large surface area, the unique porous structure, hydrophobic interaction, gate-opening effect, and enhanced properties after etching including faster mass transport, multiple active components, and more exposed active sites. The fabricated hollow ZIF-7 coated fiber lasted at least 140 cycles of extraction/desorption/aging without obvious decrease of extraction ability and no change of crystal structure. Finally, the hollow ZIF-7 coated fiber combined with GC-FID had been successfully used to detect the five biomarkers in the headspace gas of human breast cancer cell lines (MDA-MB-231) and normal mammary cell lines (CCD-1095Sk) with the recoveries of 84-105%. These results revealed the prospect of hollow MOFs as efficient adsorbents for sample pretreatment.


Asunto(s)
Neoplasias de la Mama , Zeolitas , Biomarcadores , Línea Celular , Femenino , Humanos , Microextracción en Fase Sólida , Acero Inoxidable
5.
ACS Appl Bio Mater ; 4(4): 3608-3613, 2021 04 19.
Artículo en Inglés | MEDLINE | ID: mdl-35014446

RESUMEN

Metal-organic frameworks (MOFs) are a new kind of microporous materials whose unique properties make them promising as coatings for solid phase microextraction (SPME). However, previous MOF coatings for SPME exclusively focus on single-linker MOFs, and the selective enrichment of polar or nonpolar targets depends on the polarity of linker on the surface of MOFs, which greatly limits the application of MOF coating for SPME in real samples. Here, we report a hybrid MOF-coated stainless steel fiber for SPME of biomarkers in exhaled breath from gastric cancer patients. Zeolitic imidazolate framework-8-90 (ZIF-8-90) possesses the aldehyde groups and methyl groups in the framework as a model MOF, and eight biomarkers (ethanol, acetone, hexanal, hexanol, nonane, isoprene, heptane, and decane) were used as the target analytes. The ZIF-8-90-coated fiber shows high enrichment efficiency for hydrophilic targets and hydrophobic targets, wide linearity (three orders of magnitude), and low detection limits (0.82-2.64 µg L-1). The ZIF-8-90-coated fiber exhibited higher enrichment performance for all the investigated analytes as a result of the synergy of methyl and aldehyde groups, the porous structure, and the suitable pore size of ZIF-8-90 (4-5 Å). The relative standard deviation (RSD) of six repetitions for extractions using the same ZIF-8-90-coated fiber ranged from 2.5 to 7.3%. The reproducibility between the three fibers prepared in parallel varied in the range of 4.8-12% (RSD). The fabricated ZIF-8-90-coated fiber lasted for at least 120 cycles of extraction/desorption/conditioning without an obvious reduction in extraction efficiency and precision. Finally, the developed ZIF-8-90-coated SPME fiber has been successfully used for the analysis of exhaled breath samples from gastric patients with satisfied recoveries (88-106%).


Asunto(s)
Biomarcadores/análisis , Imidazoles/química , Estructuras Metalorgánicas/química , Neoplasias Gástricas/diagnóstico , Acetona/análisis , Acetona/aislamiento & purificación , Aldehídos/análisis , Aldehídos/aislamiento & purificación , Pruebas Respiratorias/métodos , Etanol/análisis , Etanol/aislamiento & purificación , Humanos , Límite de Detección , Reciclaje , Microextracción en Fase Sólida , Neoplasias Gástricas/patología
6.
Zhongguo Zhen Jiu ; 40(4): 379-83, 2020 Apr 12.
Artículo en Chino | MEDLINE | ID: mdl-32275366

RESUMEN

OBJECTIVE: To explore the therapeutic effect and partial mechanism of electroacupuncture (EA) for patients with insulin resistance (IR) polycystic ovary syndrome (PCOS). METHODS: Seventy patients with IR-PCOS were randomly divided into an EA group (36 cases, 5 cases dropped off) and a medication group (34 cases, 4 cases dropped off). The patients in the medication group were treated with oral administration of metformin hydrochloride, 500 mg each time, twice a day. The patients in the EA group were treated with EA (continuous wave, 2 Hz of frequency) at Zusanli (ST 36), Zhongwan (CV 12), Qihai (CV 6), Yishu (EX-B 3), Shenshu (BL 23), Pishu (BL 20), Ciliao (BL 32) for 30 min, three times a week. One menstrual cycle or 4 weeks were taken as a course of treatment, and 3 continuous courses were given. The follow-up was 3 months. The lipid metabolism indexes of triacylglycerol (TG), total cholesterol (TC), high-density lipoprotein (HDL) and low-density lipoprotein (LDL), homeostasis model assessment-insulin resistance index (HOMA-IR) and testosterone (T) in serum were compared before and after treatment, and the clinical effects of the two groups were evaluated during the follow-up. RESULTS: The total effective rate was 67.7% (21/31) in the EA group and 60.0% (18/30) in the medication group, with no significant difference between the two groups (P>0.05). After treatment, the levels of serum T, HOMA-IR, LDL, TG and TC were decreased significantly in the two groups (P<0.01, P<0.05), and HDL was increased significantly (P<0.01); the levels of TC in the EA group after treatment was lower than that in the medication group (P<0.05). CONCLUSION: EA may adjust some dyslipidemia in patients to correct IR and improve endocrine disorder of PCOS, which had superior/similar effects to metformin.


Asunto(s)
Electroacupuntura , Resistencia a la Insulina , Síndrome del Ovario Poliquístico/terapia , Puntos de Acupuntura , Femenino , Humanos
7.
Mikrochim Acta ; 186(8): 588, 2019 07 31.
Artículo en Inglés | MEDLINE | ID: mdl-31367797

RESUMEN

A method is described for non-invasive glucose monitoring of diabetics by means of breath analysis. The metal-organic frameworks (MOFs) ZIF-7, UiO-66 and MOF-5 were chosen as sorbents in packed tubes for sampling and preconcentration of acetone and isopropanol which are established diabetes biomarkers. The MOF UiO-66 was found to be the most appropriate sorbent. Following thermal desorption, acetone and isopropanol where quantified by GC. The method has low limits of detection (0.79-0.84 µg·L-1) and wide linear ranges (5-2000 µg·L-1). It is assumed that the good performance of UiO-66 as a sorbent results from its large surface area and unique porous structure, and from van der Waals interactions. The relative standard deviation for six replicate cycles of sampling and preconcentration using one 50 mg UiO-66 packed tube ranged between 2.3 and 6.7% for intra-day assays, and from 2.7 to 4.3% for inter-day assays. A tube packed with 50 mg of UiO-66 packed tube can be used in over 120 cycles of adsorption/desorption without significant loss of collection efficiency. The GC method has been applied for the analysis of diabetic breath samples, and the recoveries from spiked samples ranged from 89.1 to 107.6%. Graphical abstract Schematic presentation of metal-organic frameworks as sorbents combined with thermal desorption-gas chromatography for the determination of acetone and isopropanol in exhaled breath of diabetics.


Asunto(s)
2-Propanol/análisis , Acetona/análisis , Cromatografía de Gases/métodos , Estructuras Metalorgánicas/química , 2-Propanol/química , 2-Propanol/aislamiento & purificación , Acetona/química , Acetona/aislamiento & purificación , Adsorción , Pruebas Respiratorias , Diabetes Mellitus/metabolismo , Diabetes Mellitus/patología , Humanos , Límite de Detección , Extracción en Fase Sólida
8.
Mikrochim Acta ; 185(7): 342, 2018 06 27.
Artículo en Inglés | MEDLINE | ID: mdl-29951844

RESUMEN

This review (with 85 refs.) summarizes the recent literature on the adsorption of common aromatic pollutants by using modified metal-organic frameworks (MOFs). Four kinds of aromatic pollutants are discussed, namely benzene homologues, polycyclic aromatic hydrocarbons (PAHs), organic dyes and their intermediates, and pharmaceuticals and personal care products (PPCPs). MOFs are shown to be excellent adsorbents that can be employed to both the elimination of pollutants and to their extraction and quantitation. Adsorption mechanisms and interactions between aromatic pollutants and MOFs are discussed. Finally, the actual challenges of existence and the perspective routes towards future improvements in the field are addressed. Graphical abstract Recent advance on adsorption of common aromatic pollutants including benzene series, polycyclic aromatic hydrocarbons, organic dyes and their intermediates, pharmaceuticals and personal care products by metal-organic frameworks.

9.
Mikrochim Acta ; 185(6): 307, 2018 05 22.
Artículo en Inglés | MEDLINE | ID: mdl-29789955

RESUMEN

A stainless steel fiber was coated with a gate-opening controlled metal-organic framework ZIF-7 via a sol-gel method and applied to the solid-phase microextraction of aldehydes (hexanal, heptanal, octanal, nonanal, decanal) from exhaled breath by lung cancer patients. The effects of temperature and time on the sorption and desorption were optimized. Under optimum condition, the modified fiber displays enrichment factors (typically ranging from 300 to 10,000), low limits of detection (0.61-0.84 µg L-1), and wide linear ranges of hexanal, heptanal (5-500 µg L-1) and octanal, nonanal, decanal (10-1000 µg L-1). The high extraction capability for aldehydes is thought to result from (a) the combined effects of the large surface area and the unique porous structure of the ZIF-7, (b) the hydrophobicity and gate-opening effect of the sorbent, (c) the high selectivity of the window, and (d) the presence of unsaturated metal-coordination sites. The coated fiber is thermally stable and can be re-used >150 times. The relative standard deviation (RSD) for six replicate extractions using a single fiber ranged from 1.4-15.3% for intra-day and 2.4-16.1% for inter-day. The fiber-to-fiber reproducibility for three fibers prepared in parallel was in the range of 2.4-12.6% (RSD). The method was applied to the extraction of aldehydes from real samples and to the quantitation by gas chromatography. Recoveries from spiked samples ranged from 84 to 113%. Graphical abstract A metal-organic framework ZIF-7 coated stainless steel fiber was prepared via sol-gel method. The self-made fiber was applied in the solid phase microextraction of aldehydes from exhaled breath of lung cancer patients.


Asunto(s)
Aldehídos/análisis , Aldehídos/aislamiento & purificación , Pruebas Respiratorias , Espiración , Neoplasias Pulmonares , Estructuras Metalorgánicas/química , Microextracción en Fase Sólida/métodos , Humanos , Límite de Detección , Temperatura , Factores de Tiempo
10.
J Chromatogr A ; 1343: 188-94, 2014 May 23.
Artículo en Inglés | MEDLINE | ID: mdl-24767798

RESUMEN

Metal-organic frameworks (MOFs) are attractive as porous stationary phase for open-tubular capillary electrochromatography (OT-CEC) due to their fascinating structures and unusual properties. Here we report a directly covalent bonding approach to prepare uniform and dense MOF film on the inner wall of fused silica capillary at room temperature for OT-CEC. Zeolitic imidazolate framework-90 (ZIF-90) as a model MOF because it not only possesses large surface area and high stability but also provides the free aldehyde groups to bond to the inner surface of capillary via covalent bond. X-ray diffraction, scan electron microscopy, and UV-vis spectrophotometry were used to confirm the bonding of the ZIF-90 to the inner wall of the silica capillary. The ZIF-90 coating not only increased the phase ratio of open-tubular column, but also improved the interactions of tested analytes and the coating. Owing to the porous structure of ZIF-90 and hydrophobic interactions between the analytes and the organic ligands of ZIF-90, three groups of isomers, neutral and basic compounds and nonsteroidal anti-inflammatory drugs were well separated on the ZIF-90 bonded column. The precisions (relative standard deviation, RSD) of retention time, half peak width and peak area for three consecutive runs were 0.3-1.2%, 1.3-6.0% and 1.5-5.2%, respectively. The run-to-run, day-to-day, and column-to-column precisions (RSDs) for the electroosmotic flow of the ZIF-90 bonded column were 0.2%, 0.4%, and 1.9%, respectively. Moreover, the ZIF-90 bonded column could stand more than 230 runs without observable change in the separation efficiency.


Asunto(s)
Electrocromatografía Capilar/métodos , Imidazoles/química , Electroósmosis , Concentración de Iones de Hidrógeno , Isomerismo , Microscopía Electrónica de Rastreo , Dióxido de Silicio/química , Temperatura , Difracción de Rayos X
11.
Chem Commun (Camb) ; 49(21): 2142-4, 2013 Mar 14.
Artículo en Inglés | MEDLINE | ID: mdl-23389813

RESUMEN

Here we report a covalent bonding approach to fabricate a robust metal-organic framework ZIF-90 coating for solid-phase microextraction. The ZIF-90 bonded fiber exhibits high enhancement factors, wide linearity, excellent reproducibility, and good lifetime for the studied polar endocrine disruptors.


Asunto(s)
Imidazoles/química , Dióxido de Silicio/química , Zeolitas/química , Resinas Acrílicas/química , Compuestos de Bencidrilo/análisis , Compuestos de Bencidrilo/aislamiento & purificación , Cromatografía de Gases , Dimetilpolisiloxanos/química , Disruptores Endocrinos/análisis , Disruptores Endocrinos/aislamiento & purificación , Fenoles/análisis , Fenoles/aislamiento & purificación , Polivinilos/química , Microextracción en Fase Sólida , Temperatura
12.
Zhen Ci Yan Jiu ; 37(1): 72-6, 2012 Feb.
Artículo en Chino | MEDLINE | ID: mdl-22574574

RESUMEN

Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disease in women of reproductive age, and the obesity and insulin resistance are considered to be the key link in the pathophysiological process of PCOS of obesity type. Adiponectin, a protein hormone, is closely related to insulin resistance and obesity, which has been being researched extensively in recent years. The authors of the present article review the pathogenesis of PCOS of obesity type from the relationship between adiponectin and obesity, and between adiponectin and insulin resistance, separately. In particular, the authors review studies on the underlying mechanism of acupuncture and moxibustion interventions in regulating adiponectin level briefly. The authors think of that acupuncture and moxibustion interventions induced increase of adiponectin level is possibly to improve insulin resistance in obesity and/or PCOS patients, hoping to provide a new target for clinical treatment of PCOS.


Asunto(s)
Terapia por Acupuntura , Adiponectina/genética , Obesidad/genética , Obesidad/terapia , Síndrome del Ovario Poliquístico/genética , Síndrome del Ovario Poliquístico/terapia , Adiponectina/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Moxibustión , Obesidad/metabolismo , Síndrome del Ovario Poliquístico/metabolismo
13.
Dig Dis Sci ; 57(9): 2286-95, 2012 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-22531889

RESUMEN

BACKGROUND: Previous studies have shown that moxibustion on Tianshu (ST25) and Qihai (CV6) is effective for treating Crohn's disease. However, the mechanism of moxibustion has not been clearly elucidated. AIM: The purpose of this study was to investigate the effect of moxibustion on the inhibition of colonic epithelial cell apoptosis and on tumor necrosis factor alpha (TNF-alpha) and tumor necrosis factor receptor TNF receptor-1 (TNFR1) and TNFR2 and to determine the mechanism of its protective effect using Crohn's disease (CD) model rats. METHODS AND RESULTS: The experimental CD rat models were established by the administration of trinitrobenzene sulfonic acid. In the herbs-partitioned moxibustion (HPM) and mild-warm moxibustion (MWM) groups, moxibustion was administered to Tianshu (ST25) and Qihai (CV6) acupoints once daily for 14 days. In the salicylazosulfapyridine (SASP) group, SASP was administered twice daily for 14 days. A normal control (NC) group and a model control (MC) group were also studied. The levels of TNF-alpha and its mRNA, TNFR1 as well as the rate of colonic epithelial cell apoptosis were significantly decreased in the HPM, MWM and SASP groups compared with the MC group. The HPM and MWM groups had lower mRNA expression and lower protein levels of TNF-alpha compared to the SASP group. The HPM and MWM groups exhibited less apoptosis than the SASP group. CONCLUSIONS: Moxibustion may inhibit colonic epithelial cell apoptosis by reducing the high expression of TNF-alpha and TNFR1 to protect the defective colonic epithelial barrier in CD model rats.


Asunto(s)
Colon/metabolismo , Enfermedad de Crohn/terapia , Mucosa Intestinal/metabolismo , Moxibustión/métodos , Receptores Tipo I de Factores de Necrosis Tumoral/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Apoptosis , Colon/patología , Citometría de Flujo , Regulación de la Expresión Génica , Etiquetado Corte-Fin in Situ , Ratas , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Factor de Necrosis Tumoral alfa/genética
14.
World J Gastroenterol ; 17(45): 4960-70, 2011 Dec 07.
Artículo en Inglés | MEDLINE | ID: mdl-22174545

RESUMEN

AIM: To investigate the effects of moxibustion on down-regulation of the colonic epithelial cell apoptosis and repair of the tight junctions in rats with Crohn's disease (CD). METHODS: Sixty male Sprague-Dawley rats were randomly divided into a normal control (NC) group, a model control (MC) group, an herbs-partitioned moxibustion (HPM) group, a mild-warm moxibustion (MWM) group and a salicylazosulphapyridine (SASP) group, with 12 rats in each group. The CD model rats were treated with trinitrobenzene sulphonic acid to induce intestinal inflammation. The rats in the HPM and MWM groups were treated at the Tianshu (ST25) and Qihai (CV6) acupoints once daily for 14 d, and the SASP group was fed SASP twice daily for 14 d. No additional treatment was given to the MC and NC groups. The microstructure of the colonic epithelium was observed under a transmission electron microscope, the transepithelial resistance was measured using a short-circuit current, colonic epithelial cell apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling assay, and the expression of occludin, claudin-1 and zonula occludens-l (ZO-1) in the colonic epithelial junction was determined by Western blotting and immunofluorescence staining. RESULTS: Compared with the MC group, the microstructure of the colonic epithelial barrier was significantly improved in rats treated with HPM, MWM or SASP, meanwhile, the current flow was reduced significantly, with values of 168.20 ± 6.14 vs 99.70 ± 3.13, 99.10 ± 4.28 and 120.30 ± 3.65 mA, respectively (P = 0.001). However, the HPM and MWM groups had higher current flow rates than the SASP group (99.70 ± 3.13, 99.10 ± 4.28 vs 120.30 ± 3.65 mA, P = 0.001). The number of the apoptotic colonic epithelial cells in HPM, MWM and SASP groups was largely reduced (61.5 ± 16.91 vs 15.5 ± 8.89, 14.8 ± 6.27 and 24.7 ± 9.68, respectively (P = 0.001); and the expression of occludin, claudin-1 and ZO-1 in the MWM and HPM groups was significantly enhanced (0.48 ± 0.10, 0.64 ± 0.09 vs 0.18 ± 0.05 for occludin, 0.12 ± 0.02, 0.17 ± 0.03 vs 0.05 ± 0.01 for claudin-1, and 0.08 ± 0.01, 0.11 ± 0.01 vs 0.02 ± 0.01 for ZO-1). And in SASP group, the expression of occludin and ZO-1 was also significantly increased (0.27 ± 0.04 vs 0.18 ± 0.05 for occludin and 0.05 ± 0.01 vs 0.02 ± 0.01 for ZO-1), but there was no significant difference for claudin-1. The HPM and MWM groups had higher expression of occludin, claudin-1 and ZO-1 than the SASP group. CONCLUSION: HPM and MWM treatment can down-regulate apoptosis of colonic epithelial cells, repair tight junctions and enhance colonic epithelial barrier function in rats with CD.


Asunto(s)
Apoptosis/fisiología , Colon/patología , Enfermedad de Crohn/patología , Enfermedad de Crohn/terapia , Células Epiteliales/fisiología , Moxibustión , Uniones Estrechas/metabolismo , Animales , Claudina-1 , Colon/citología , Regulación hacia Abajo , Células Epiteliales/ultraestructura , Humanos , Mucosa Intestinal/citología , Mucosa Intestinal/patología , Masculino , Proteínas de la Membrana/metabolismo , Ocludina , Fosfoproteínas/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Proteína de la Zonula Occludens-1
15.
Zhen Ci Yan Jiu ; 36(4): 235-41, 2011 Aug.
Artículo en Chino | MEDLINE | ID: mdl-21942174

RESUMEN

OBJECTIVE: To observe the effect of moxibustion on colonic tumor necrosis factor (TNF)-alpha level in Crohn's Disease (CD) rats and the effect of colonic supernatant of CD rats experiencing moxibustion on the expression of the tight junction proteins ocoludin, claudin-1 and zonula occiludens (ZO)-1 and their genes in the cultivated colonic epithelial cells derived from CD rats, so as to reveal its underlying mechanism in resisting colonic epithelial barrier defects. METHODS: Sixty SD rats were randomized into normal control, model, moderately warm moxibustion (MWM), herbs-partitioned moxibustion (HPM) and medication (salazosulfapyridine, SASP) groups (n=12). CD model was established by intra-annual perfusion of trinitrobenzene sulfonic acid (TNBS) solution (TNBS: 50% alcohol = 2:1, 0.5 mL/kg). For rats of the HPM and MWM groups, moxibustion was given to "Tianshu" (ST 25) and "Qihai" (CV 6) once daily for 14 d. For rats of the medication group, intragastric perfusion of SASP solution (0. 0405 g/3 mL) was given twice daily for 14 d. After the treatment, all the rats including those of normal group were killed for preparing the supernatant of colonic mucosa tissue (6-8 cm superior to the anus). The colonic epithelial cells of the normal group were purified and cultivated in DMEM culture fluid containing the prepared supernatant of normal group to establish an intestinal epi-thelial barrier defect model, and also cultured separately in the media containing the prepared supernatants of the model, medication, HPM and MWM groups. One week after the culture, the expression levels of occludin, claudin-1 and ZO-1 proteins and their genes in the cultured colonic epithelial cells were detected by Western blot and fluorescent quantitative polymerase chain reaction assay respectively. TNF-a content of the colonic supernatant was detected by enzyme linked immunosorbent assay. RESULTS: Compared with the normal group, colonic TNF-alpha content was remarkably increased in the model group (P < 0.01). In comparison with the model group, colonic TNF-acx contents were significantly decreased in the medication, MWM and HPM groups (P < 0.01), and those of the MWM and HPM groups were markedly lower than that of the medication group (P < 0.05). The expression levels of the cultured normal colonic epithelial occludin, claudin-1 and ZO-1 proteins and their mRNAs in the medication, MWM and HPM groups were remarkably increased compared with those in the model group (P < 0.01, P < 0.05). The expression levels of colo-nic epithelial occludin, claudin-1 and ZO-1 proteins and their mRNAs were significantly higher in the MWM and HPM groups than in the medication group (P < 0.05). CONCLUSION: Both MWM and HPM can downregulate colonic mucosal TNF-alpha content in CD rats, and the colonic supernatant of rats undergoing MWM and HPM may upregulate the expression of colonic epithelial occludin, claudin-1 and ZO-1 proteins and their mRNAs in the cultivated colonic epithelial cells, which may contribute to the effect of moxibustion in relieving colonic epithelial barrier defect.


Asunto(s)
Colon/química , Enfermedad de Crohn/terapia , Proteínas de la Membrana/análisis , Moxibustión , Fosfoproteínas/análisis , Factor de Necrosis Tumoral alfa/análisis , Animales , Células Cultivadas , Claudina-1 , Colon/metabolismo , Enfermedad de Crohn/metabolismo , Células Epiteliales/química , Células Epiteliales/metabolismo , Mucosa Intestinal/química , Mucosa Intestinal/metabolismo , Masculino , Proteínas de la Membrana/genética , Ocludina , Fosfoproteínas/genética , ARN Mensajero/análisis , Ratas , Ratas Sprague-Dawley , Proteína de la Zonula Occludens-1
16.
Zhonghua Yi Xue Za Zhi ; 83(6): 494-7, 2003 Mar 25.
Artículo en Chino | MEDLINE | ID: mdl-12887765

RESUMEN

OBJECTIVE: To study the molecular basis of preventive effect of human apolipoprotein-1 (h-apoA-1) on vascular smooth muscle cell (vSMC) proliferation and lipid deposition induced by oxidized low density lipoprotein (ox-LDL). METHODS: Smooth muscle cells originated from the aortae of h-apo-A-1 transgenic mice were cultured and divided into 2 groups, one group was stimulated by ox-LDL (tester) and the other group was used as control (driver). Subtractive hybridization was used to enrich the genes differentially expressed in the vSMCs induced by ox-LDL. A subtractive library was thus established and confirmed by colony hybridization in situ and dot blot analysis. 15 clones out of the 57 differentially expressed clones were randomly chosen foe sequencing and homology analysis. The whole-length cDNA library of vSMC induced by ox-LDL was established using SMART technique. RESULTS: Three expression sequence tags (EST), all correlated with immune system, were confirmed: C1-inhibitor (C1-INH), lectin, and T cell receptor beta. The whole-length cDNA library contained 1.5 x 10(6) pfu/ml primary recombinants with insertions 0.5 - 3 kb in length. CONCLUSION: The 3 EST may be involved in the mechanism of atherogenesis by ox-LDL and the mechanism of the function of h-ApoA-1 in retarding the progression of atherogenesis induced by ox-LDL.


Asunto(s)
Apolipoproteína A-I/fisiología , Arteriosclerosis/etiología , Metabolismo de los Lípidos , Lipoproteínas LDL/toxicidad , Músculo Liso Vascular/citología , Animales , Apolipoproteína A-I/genética , División Celular , Etiquetas de Secuencia Expresada , Expresión Génica , Humanos , Ratones , Ratones Transgénicos , Músculo Liso Vascular/metabolismo
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