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1.
Chin J Physiol ; 63(1): 27-34, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32056984

RESUMEN

Three-quarters of the lands in Taiwan are over 1000 m above sea level. Formosan wood mice (Apodemus semotus), also called Taiwanese field mice, are largely found at altitudes of 1400 ~ 3700 m and are the dominant rodents in these areas. Notably, Formosan wood mice show high levels of exploratory behaviors, not only in the wild but also in laboratory situations. Therefore, in this study, we examined the behavioral responses and central dopaminergic activities of male C57BL/6J mice and Formosan wood mice in the open field test. Dopamine and its major metabolite 3,4-dihydroxyphenylacetic acid were used as indices of dopaminergic activities. Formosan wood mice showed higher levels of exploration and locomotor activity than C57BL/6J mice in the open field test. Higher central dopaminergic activities in the nucleus accumbens, striatum, and medial prefrontal cortex were found in Formosan wood mice than in C57BL/6J mice in the open field test. Higher levels of locomotion and central dopaminergic activities in Formosan wood mice were consistent after two exposures to the open field test; however, dramatic decreases in levels of locomotion and central dopaminergic activities in C57BL/6J mice were found after two exposures to the open field test. The present study found that Formosan wood mice exhibited higher levels of locomotor activity and exploration and central dopaminergic activities than C57BL/6J mice after one or two exposures to the open field test.


Asunto(s)
Conducta Exploratoria , Animales , Dopamina , Masculino , Ratones , Ratones Endogámicos C57BL , Murinae , Taiwán
2.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(1): 29-33, 2020 Feb.
Artículo en Chino | MEDLINE | ID: mdl-32027249

RESUMEN

OBJECTIVE: To explore whether BAX plays a role in the development of Philadelphia chromosome-positive leukemia and related mechanisms. METHODS: Target-gene knockout mice were used as bone marrow cell donors. Retrovirus over-expressing BCR-ABL were packaged. BCR-ABL-induced B-ALL mouse model was established through donor's B cells transfected by the retrovirus and the B cells over-expressing BCR-ABL were given to the receptor mice by tail vein injection. Western blot was used to detect the protein express and flow cytometry was used to analyze the B cell subpopulations in BAX-/- and WT mouse bone marrows. Kaplan-Meier analysis was used to estimate the survival of diseased mice. RESULTS: BAX deletion caused faster development of BCR-ABL-induced leukemia in vitro and in vivo. BCR-ABL increased BCL-2 expression and enhanced BCL-2/BAX heterodimer formation. CONCLUSION: The BAX deletion can accelerate the disease progression of BCR-ABL induced B-ALL.


Asunto(s)
Leucemia Mielógena Crónica BCR-ABL Positiva , Animales , Células de la Médula Ósea , Progresión de la Enfermedad , Proteínas de Fusión bcr-abl , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Ratones , Proteína X Asociada a bcl-2
3.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(1): 242-247, 2020 Feb.
Artículo en Chino | MEDLINE | ID: mdl-32027284

RESUMEN

OBJECTIVE: To investigate the preventive and therapeutic effects of endothelial progenitor cells on monocrotaline-induced hepatic vein occlusion disease in mice. METHODS: C57BL/6 mice were randomly divided into 3 groups: saline group (n=15), monocrotaline group (n=15), and endothelial progenitor cell infusion group (n=15). Liver function (TBIL, ALT, AST), liver index, and serum levels of TNF-α and IL-6 were measured on the 8th day after intragastric administration. Hepatic sinusoidal endothelial cells, hepatic central venous endothelial cells and hepatocytes were observed by both HE and immunohistochemical staining. Hepatic fibrosis was observed by Masson's trichrome staining. RESULTS: By the light microscopy, the liver of the monocrotaline group showed moderate to the severe injuries of hepatic sinusoidal and central venous endothelial cells, and hepatic venous congestion. Masson staining showed moderate to severe hepatic fibrosis of central vein and hepatic sinus. In the endothelial progenitor cell group, hepatic sinusoidal and central venous endothelial cell injuries, and the fibrosis of central hepatic vein and hepatic sinus were mild to moderate. Hepatic venous congestion was reduced in comparison with that in the mice of the monocrotaline group. Compared with the endothelial progenitor cell group, the liver index was higher, the liver function was more abnormal, and the serum expression levels of TNF-α and IL-6 were higher in the monocrotaline group. CONCLUSION: The monocrotaline-induced damage of hepatic sinusoidal and central venous endothelial cells is an linitiating factor for hepatic vein occlusive disease. Infusion of endothelial progenitor cells can play a role in preventing and treating hepatic vein occlusion.


Asunto(s)
Células Progenitoras Endoteliales , Enfermedad Veno-Oclusiva Hepática , Animales , Venas Hepáticas , Hígado , Ratones , Ratones Endogámicos C57BL , Monocrotalina
4.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(1): 267-274, 2020 Feb.
Artículo en Chino | MEDLINE | ID: mdl-32027288

RESUMEN

OBJECTIVE: To investigate the effects of human amniotic mesenchymal stem cell(AMSC) on acute graft-versus-host disease (aGVHD) in xenotransplatation. METHODS: NPG mice were injected with human PBMNC via tail vein to establish a xenografted aGVHD model. The mice in the experimental group were divided into PBMNC infusion group and PBMNC+AMSC co-infusion group, the general condition, survival time and manifestations of aGVHD were observed, the body weight and blood routine indicators were detected, the pathological changes of aGVHD target organs (lung, liver, spleen, small intestine) were observed by HE staining, and the levels of human T cells in peripheral blood, tissues and organs of mice was detected by flow cytometry. RESULTS: The manifestations of aGVHD (lassitude hunchback, shrub, weight reduction, etc.) and the pathological damage of the target organs (lung, liver, spleen, intestine) in PBMNC+AMSC co-infusion group were lighter than those in PBMNC infusion group. Moreover, the PBMNC and AMSC co-infusion significantly reduced the implantion proportion of human T lymphocytes (CD3+, CD45+) in mice and increased the ratio of CD4+/CD8+. CONCLUSION: Infusion of human-derived AMSC can attenuate the manifestations of aGVHD in mouse xenografts to a certain level, and improve the pathological damage of receptor target organs.


Asunto(s)
Enfermedad Injerto contra Huésped , Células Madre Mesenquimatosas , Enfermedad Aguda , Animales , Xenoinjertos , Humanos , Ratones , Linfocitos T , Trasplante Heterólogo
5.
Zhonghua Xin Xue Guan Bing Za Zhi ; 48(1): 61-65, 2020 Jan 24.
Artículo en Chino | MEDLINE | ID: mdl-32008297

RESUMEN

Objective: To investigate the feasibility of echocardiography-guided closed-chest repeated intraventricular blood sampling in mice, and to clarify the maximum blood volume that can be collected by this method, and whether the method can be used for long-term repeated blood collection in mice. Methods: Twenty-four male C57BL/6J mice (10-14 weeks old) were divided into the terminal experiment group (n=4, for investigating the maximum blood amount that could be sampled at one time), the repeated 0.5 ml blood collection group (n=10, sampling 0.5 ml whole blood each time, once every two days for consecutive 4 weeks), and the repeated 0.75 ml blood collection group (n=10, sampling 0.75 ml whole blood each time, once every two days for consecutive 4 weeks). High-frequency echocardiography was used to display the largest section of the left ventricle, guiding the insulin syringe needle through the thorax into the left ventricle for blood collection. In the repeated 0.5 ml blood collection group, echocardiography was used to detect the cardiac structure and function before blood collection, three minutes after blood collection, and one week after the last (the 14th) blood collection. Results: We successfully performed echocardiography-guided closed-chest intraventricular blood sampling, with an average operating time (88±19)s per mouse, and a maximum blood volume (1.43±0.11)ml per mouse. In the repeated 0.5 ml blood collection group, heart rate, left ventricular ejection fraction, left ventricular fractional shortening, left ventricular end-diastolic dimension and left ventricular posterior wall end-diastolic thickness remained uncganged before the first blood collection and after 4 weeks of repeated blood collection (all P>0.05). No death in the repeated 0.5 ml blood collection group. However, in the 0.75 ml blood collection group, two mice died before the end point. Conclusions: The echocardiography-guided closed-chest intraventricular blood sampling is a safe, minimally invasive, convenient and efficient method, and can be used repeatedly for long-term blood collection in mice.


Asunto(s)
Ecocardiografía , Ventrículos Cardíacos , Animales , Estudios de Factibilidad , Masculino , Ratones , Ratones Endogámicos C57BL , Función Ventricular Izquierda
6.
Cell Physiol Biochem ; 54(1): 126-141, 2020 Feb 05.
Artículo en Inglés | MEDLINE | ID: mdl-32017483

RESUMEN

BACKGROUND/AIMS: Serotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter and hormone with important physiological functions in many organs, including the intestine. We have previously shown that 5-HT activates the aryl hydrocarbon receptor (AhR) in intestinal epithelial cells (IECs) via a serotonin transporter (SERT)-dependent mechanism. AhR is a nuclear receptor that binds a variety of molecules including tryptophan (TRP) metabolites to regulate physiological processes in the intestine including xenobiotic detoxification and immune modulation. We hypothesized that 5-HT activates AhR indirectly by interfering with metabolic clearance of AhR ligands by cytochrome P450 1A1 (CYP1A1). METHODS: Inhibition of CYP1A1 activity by 5-HT was assessed in the human intestinal epithelial cell line Caco-2 and recombinant CYP1A1 microsomes using both luciferase and LC-MS/MS. Degradation of 5-HT by recombinant CYP1A1 was measured by LC-MS/MS. For in vitro studies, CYP1A1 and CYP1B1 mRNA expression levels were measured by RT-PCR and CYP1A1 activity was measured by ethoxyresorufin-O-deethylase (EROD) assays. For in vivo studies, AhR ligands were administered to SERT KO mice and WT littermates and intestinal mucosa CYP1A1 mRNA was measured. RESULTS: We show that 5-HT inhibits metabolism of both the pro-luciferin CYP1A1 substrate Luc-CEE as well as the high affinity AhR ligand 6-formylindolo[3,2-b] carbazole (FICZ). Recombinant CYP1A1 assays revealed that 5-HT is metabolized by CYP1A1 in an NADPH dependent manner. Treatment with 5-HT in TRP-free medium, which is devoid of trace AhR ligands, showed that 5-HT requires the presence of AhR ligands to activate AhR. Cotreatment with 5-HT and FICZ confirmed that 5-HT potentiates induction of AhR target genes by AhR ligands. However, this was only true for ligands which are CYP1A1 substrates such as FICZ. Administration of ß-napthoflavone by gavage or indole-3-carbinol via diet to SERT KO mice revealed that lack of SERT impairs intestinal AhR activation. CONCLUSION: Our studies provide novel evidence of crosstalk between serotonergic and AhR signaling where 5-HT can influence the ability of AhR ligands to activate the receptor in the intestine.


Asunto(s)
Citocromo P-450 CYP1A1/metabolismo , Receptores de Hidrocarburo de Aril/metabolismo , Serotonina/farmacología , Transcripción Genética/efectos de los fármacos , Animales , Células CACO-2 , Carbazoles/farmacología , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1B1/genética , Citocromo P-450 CYP1B1/metabolismo , Humanos , Ligandos , Masculino , Ratones , Ratones Noqueados , ARN Mensajero/metabolismo , Receptores de Hidrocarburo de Aril/química , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas de Transporte de Serotonina en la Membrana Plasmática/deficiencia , Proteínas de Transporte de Serotonina en la Membrana Plasmática/genética , Transducción de Señal/efectos de los fármacos , Especificidad por Sustrato , beta-naftoflavona/administración & dosificación
7.
Zhonghua Xue Ye Xue Za Zhi ; 41(1): 34-39, 2020 Jan 14.
Artículo en Chino | MEDLINE | ID: mdl-32023752

RESUMEN

Objective: To evaluate the effects of adeno-associated virus (AAV) carrying hFⅧ by serotype 8 (AAV8/hFⅧ) on hemophilia A (HA) mice by gene therapy strategy. Methods: pAAV-CB-EGFP, pH22 (serotype 2) and pfΔ6 (adenovirus helper) were used to package AAV into HEK-293 cells in different conditions (ratios of cells to plasmids). The efficiency of transfection and infection were evaluated using immunofluorescence microscope to seek an optimized package condition. pAAV-TTR-hFⅧ, pH 28 (serotype 8) and pfΔ6 were applied to package AAV8/hFⅧ in HEK-293 cells using the optimized package condition. The purified AAV8/hFⅧ were intravenously injected into HA mice and the effects of gene therapy were estimated. Results: The efficiency of package was evaluated according to the amount and intensity of enhanced green fluorescent protein (EGFP) under immunofluorescence microscope. Four package conditions including 10 cm-dish to transfect 10 µg plasmids, 20 cm-dish to 20 µg, 30 µg and 40 µg plasmids were employed, and the condition of 20 cm-dish to transfect 20 µg plasmids reached the highest transfection efficiency at 24 h, 48 h and 72 h after transfection. The small scale AAV-EGFP was packaged using the optimized condition and an AAV crude extract was harvested by a freeze-thaw method. HEK-293 and 16095 cells were infected by the AAV crude extract, and the preferential infection efficiency was recognized in 16095 cells under immunofluorescence microscope. Then, AAV8/hFⅧ was packaged and purified based on the optimized transfection condition, and the high purity of AAV8/hFⅧ was detected by Western blot. Fractions of AAV8/hFⅧ at the dose of 8×10(12) vg/kg were injected into HA mice through tail vein, an eye-bleeding was performed at every two weeks, and the activity of FⅧ was measured by aPTT assay. Results showed that the activity of FⅧ maintained at the therapeutic level and lasted up to 12 weeks after injection. Conclusion: The purified AAV8/hFⅧ based on the optimized package condition could play a role in HA mice gene therapy, and the long-term therapeutic effects of AAV8/hFⅧ were observed in vivo.


Asunto(s)
Dependovirus , Hemofilia A , Animales , Terapia Genética , Vectores Genéticos , Células HEK293 , Humanos , Ratones
8.
Int J Oral Sci ; 12(1): 5, 2020 02 06.
Artículo en Inglés | MEDLINE | ID: mdl-32024813

RESUMEN

Rheumatoid arthritis (RA) is an autoimmune disease affecting 1% of the world population and is characterized by chronic inflammation of the joints sometimes accompanied by extra-articular manifestations. K/BxN mice, originally described in 1996 as a model of polyarthritis, exhibit knee joint alterations. The aim of this study was to describe temporomandibular joint (TMJ) inflammation and damage in these mice. We used relevant imaging modalities, such as micro-magnetic resonance imaging (µMRI) and micro-computed tomography (µCT), as well as histology and immunofluorescence techniques to detect TMJ alterations in this mouse model. Histology and immunofluorescence for Col-I, Col-II, and aggrecan showed cartilage damage in the TMJ of K/BxN animals, which was also evidenced by µCT but was less pronounced than that seen in the knee joints. µMRI observations suggested an increased volume of the upper articular cavity, an indicator of an inflammatory process. Fibroblast-like synoviocytes (FLSs) isolated from the TMJ of K/BxN mice secreted inflammatory cytokines (IL-6 and IL-1ß) and expressed degradative mediators such as matrix metalloproteinases (MMPs). K/BxN mice represent an attractive model for describing and investigating spontaneous damage to the TMJ, a painful disorder in humans with an etiology that is still poorly understood.


Asunto(s)
Artritis Experimental/patología , Artritis Reumatoide/patología , Huesos/diagnóstico por imagen , Articulación Temporomandibular/diagnóstico por imagen , Articulación Temporomandibular/lesiones , Microtomografía por Rayos X/métodos , Animales , Artritis Experimental/inmunología , Artritis Reumatoide/inmunología , Huesos/metabolismo , Huesos/patología , Modelos Animales de Enfermedad , Humanos , Imagen por Resonancia Magnética , Metaloproteinasa 8 de la Matriz/inmunología , Ratones , Ratones Transgénicos , Articulación Temporomandibular/metabolismo , Tomografía Computarizada por Rayos X
9.
Cell Physiol Biochem ; 54(1): 142-159, 2020 Feb 07.
Artículo en Inglés | MEDLINE | ID: mdl-32028545

RESUMEN

BACKGROUND/AIMS: It is well established that oxidative stress and inflammation are common pathogenic features of retinal degenerative diseases. ITH12674 is a novel compound that induces the transcription factor Nrf2; in so doing, the molecule exhibits anti-inflammatory, and antioxidant properties, and affords neuroprotection in rat cortical neurons subjected to oxidative stress. We here tested the hypothesis that ITH12674 could slow the retinal degeneration that causes blindness in rd10 mice, a model of retinitis pigmentosa. METHODS: Animals were intraperitoneally treated with 1 or 10 mg/Kg ITH12674 or placebo from P16 to P30. At P30, retinal functionality and visual acuity were analyzed by electroretinography and optomotor test. By immunohistochemistry we quantified the photoreceptor rows and analyzed their morphology and connectivity. Oxidative stress and inflammatory state was studied by Western blot, and microglia reactivity was monitored by flow cytometry. The blood-brain barrier permeation of ITH12674 was evaluated using a PAMPA-BBB assay. RESULTS: In rd10 mice treated with 10 mg/Kg of the compound, the following changes were observed (with respect to placebo): (i) a decrease of vision loss with higher scotopic a- and b-waves; (ii) increased visual acuity; (iii) preservation of cone photoreceptors morphology, as well as their synaptic connectivity; (iv) reduced expression of TNF-α and NF-κB; (v) increased expression of p38 MAPK and Atg12-Atg5 complex; and (vi) decreased CD11c, MHC class II and CD169 positive cell populations. CONCLUSION: These data support the view that a Nrf2 inducer compound may arise as a new therapeutic strategy to combat retinal neurodegeneration. At present, we are chemically optimising compound ITH12674 with the focus on improving its neuroprotective potential in retinal neurodegenerative diseases.


Asunto(s)
Isotiocianatos/uso terapéutico , Melatonina/análogos & derivados , Factor 2 Relacionado con NF-E2/agonistas , Retinitis Pigmentosa/tratamiento farmacológico , Animales , Barrera Hematoencefálica/efectos de los fármacos , Barrera Hematoencefálica/metabolismo , Modelos Animales de Enfermedad , Electrorretinografía , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Isotiocianatos/química , Isotiocianatos/farmacología , Masculino , Melatonina/química , Melatonina/farmacología , Melatonina/uso terapéutico , Ratones , Ratones Endogámicos C57BL , Factor 2 Relacionado con NF-E2/metabolismo , Células Fotorreceptoras/efectos de los fármacos , Células Fotorreceptoras/patología , Retina/efectos de los fármacos , Retina/metabolismo , Retinitis Pigmentosa/metabolismo , Retinitis Pigmentosa/patología , Factor de Necrosis Tumoral alfa/metabolismo , Agudeza Visual/efectos de los fármacos , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
10.
Zhonghua Zhong Liu Za Zhi ; 42(1): 30-36, 2020 Jan 23.
Artículo en Chino | MEDLINE | ID: mdl-32023766

RESUMEN

Objective: To investigate the effects of miR-513a-3p on proliferation, migration and invasion of gastric cancer cells and its mechanism. Methods: The miR-NC (miR-negative control mimics), miR-513a-3p (miR-513a-3p mimics), anti-miR-NC, anti-miR-513a-3p, si-NC, si-MDM2 (murine double minute 2), miR-513a-3p+ pcDNA3.1 (co-transfected with miR-513a-3p and pcDNA3.1), miR-513a-3p+ pcDNA3.1-MDM2 (co-transfected with miR-513a-3p and pcDNA3.1-MDM2) were transfected into BGC-823 cells, respectively. The expression of miR-513a-3p was detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), and the protein expressions of cyclin D1, MMP-2, p21, E-cadherin, MDM2 were detected by western blot. The viability of BGC-823 cells of each group was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The migration and invasion of each group were detected by Transwell, the targeting relationship between miR-513a-3p and MDM2 was detected by double luciferase reporter gene assay. Results: The expression of miR-513a-3p in gastric epithelial cells GES-1 was 0.76±0.08, significantly higher than 0.21±0.02 in gastric cancer cells BGC-823 and 0.34±0.03 in MGC-803, respectively (P<0.05). The cell viabilities of the miR-NC group at 24 h, 48 h and 72 h were 0.57±0.05, 1.03±0.10, 1.43±0.14, respectively, while those of the miR-513a-3p group were 0.36±0.03, 0.48±0.05, and 0.63±0.06, respectively. The migration and invasion numbers of miR-NC group were 130±11.80 and 117±10.60, respectively, those of miR-513a-3p group were 58±5.64 and 50±5.13, respectively, and the differences were statistically significant (P<0.05). The cell viabilities of the si-NC group at 24 h, 48 h and 72 h were 0.53±0.05, 0.95±0.10, 1.36±0.14, respectively. Those of the si-MDM2 group were 0.39±0.04, 0.57±0.06, and 0.80±0.08, respectively. The cell migration and invasion of the si-NC group were 141±12.02 and 109±10.60, respectively, while those of the MDM2 group were 66±6.67 and 61±6.18, respectively, and the differences were statistically significant (P<0.05). The cell viabilities of the miR-513a-3p+ pcDNA3.1 group at 24 h, 48 h and 72 h were 0.34±0.03, 0.46±0.05, and 0.61±0.06, respectively. Those of miR-513a-3p+ pcDNA3.1-MDM2 group were 0.48±0.05, 0.82±0.08, 1.17±0.12, respectively. The migration and invasion of miR-513a-3p+ pcDNA3.1 group were 56±5.71 and 51±5.16, respectively, while those of miR-513a-3p+ pcDNA3.1-MDM2 group were 113±10.28 and 104±10.02, respectively, and the differences were statistically significant (P<0.05). Conclusion: miR-513a-3p may inhibit the proliferation, migration and invasion of gastric cancer cells through targeting regulation of MDM2, which will provide new targets for the prevention and treatment of gastric cancer.


Asunto(s)
Proliferación Celular , MicroARNs , Neoplasias Gástricas , Animales , Línea Celular Tumoral , Movimiento Celular , Regulación Neoplásica de la Expresión Génica , Ratones , Invasividad Neoplásica , Neoplasias Gástricas/genética , Neoplasias Gástricas/patología
11.
Rinsho Ketsueki ; 61(1): 3-10, 2020.
Artículo en Japonés | MEDLINE | ID: mdl-32023599

RESUMEN

Recently, monocyte-derived fibroblast-like cells, called fibrocytes, garnered attention as involved in the novel pathogenesis of various fibrotic diseases. They also play a role in the induction of myelofibrosis (MF). Neoplastic fibrocytes are overrepresented in the bone marrow of patients with primary MF, and the suppression of fibrocyte differentiation by serum amyloid P has been shown to remarkably improve MF. Further, thrombopoietin (TPO) or a TPO receptor agonist directly induces fibrocyte differentiation, and fibrocyte elimination reversed the MF phenotype in a murine model. Human fibrocytes highly express signaling lymphocytic activation molecule-F7 (SLAMF7) compared with macrophages. Myeloproliferative neoplasm (MPN) patients harboring JAK2V617F with MF had a significantly elevated SLAMF7high monocyte percentage, which correlated positively with the JAK2V617F allele burden. Furthermore, the JAK2V617F allele burden and the tendency to differentiate into fibrocytes of SLAMF7high monocytes was significantly higher than that of JAK2V617Flow monocytes and could be a potential target of elotuzumab (Elo), an anti-SLAMF7 antibody used to treat multiple myeloma. Elo independently inhibited the differentiation of fibrocytes derived not only from healthy controls but also from MF patients in vitro. Elo also ameliorated MF and splenomegaly induced by romiplostim administration in humanized NOG mice. Thus, Elo could be a therapeutic agent for MPN patients harboring JAK2V617F with MF.


Asunto(s)
Trastornos Mieloproliferativos , Mielofibrosis Primaria , Animales , Médula Ósea , Humanos , Macrófagos , Ratones , Monocitos
12.
Zhonghua Shao Shang Za Zhi ; 36(1): 48-53, 2020 Jan 20.
Artículo en Chino | MEDLINE | ID: mdl-32023718

RESUMEN

Objective: To investigate the effects of sodium butyrate on intestinal barrier of the severe scald mice and the related mechanism. Methods: Eighteen C57BL/6 female mice, aged eight to twelve weeks, were divided into sham scald group, pure scald group, and scald+ sodium butyrate group according to random number table, with 6 mice in each group. Back of each mouse in pure scald group and scald+ sodium butyrate group were immersed into 90 ℃ water for 9 s, causing full-thickness scald of 30% total body surface area, while back of each mouse in sham scald group were immersed into 37 ℃ water for 9 s, causing sham injury. All of the mice in 3 groups were intraperitoneally injected with 1 mL sterile lactated Ringer's solution immediately after injury. Besides, mice in scald+ sodium butyrate group were intraperitoneally injected with 300 mg/kg sodium butyrate at 30 min before injury and immediately after injury, while mice in sham scald group and pure scald group were intraperitoneally injected with the same volume of sterile phosphate buffer solution. At post injury hour (PIH) 24, portal vein of mice in 3 groups was harvested, intestinal permeability was measured by fluorescin isothiocyanate-dextran fluorescence probe tracing method, then lileal tissue of mice in 3 groups was harvested, protein expressions of zonula occludens l (ZO-1), occludin, claudin-1, claudin-2, nucleotide-binding oligomerization domain-containing protein-like receptor family pyrin domain containing 3 (NLRP3), interleukin-1ß (IL-1ß), and IL-18 were detected by Western blotting, and distribution of ZO-1 in intestinal mucosa was observed by indirect immunofluorescence. Data were processed with one-way analysis of variance, least-significant difference test, and Bonferroni correction. Results: (1) At PIH 24, the intestinal permeability of mice in sham scald group, pure scald group, and scald+ sodium butyrate group was 0.88±0.19, 2.62±0.48, 1.23±0.16, respectively. Compared with that in sham scald group, the intestinal permeability of mice in pure scald group was significantly elevated (P<0.01), while the intestinal permeability of mice in scald+ sodium butyrate group showed no obvious change (P>0.05). Compared with that in pure scald group, the intestinal permeability of mice in scald+ sodium butyrate group was significantly decreased (P<0.01). (2) At PIH 24, compared with those in sham scald group, the protein expressions of ZO-1, occludin, and claudin-1 of mice in pure scald group and scald+ sodium butyrate group were significantly decreased (P<0.05), while the protein expression of claudin-2 was significantly increased (P<0.05). At PIH 24, compared with those of pure scald group, the protein expressions of ZO-1 and occludin of mice in scald+ sodium butyrate group were significantly elevated (P<0.05), while the protein expression of claudin-2 was significantly decreased (P<0.05), the protein expression of claudin-1 showed no significant difference (P>0.05). (3) At PIH 24, compared with those in sham scald group, the protein expressions of NLRP3, IL-1ß, and IL-18 of mice in pure scald group and scald+ sodium butyrate group were significantly increased (P<0.05). Compared with those of pure scald group, the protein expressions of NLRP3, IL-1ß, and IL-18 of mice in scald+ sodium butyrate group were significantly decreased (P<0.05). (4) At PIH 24, ZO-1 in intestinal mucosa of mice in sham scald group was distributed smoothly, continuously and homogeneously along the membrane. ZO-1 in intestinal mucosa of mice in pure scald group was distributed unsmoothly with breaks. The distribution of ZO-1 in intestinal mucosa of mice in scald+ sodium butyrate group was ameliorated compared with that in pure scald group. Conclusions: Sodium butyrate can inhibit the activation of NLRP3 inflammasome and decrease the production of IL-1ß and IL-18 in intestinal mucosa of severe scald mice, which protects the intestinal barrier function by alleviating the alteration of tight junction protein expression and localization.


Asunto(s)
Quemaduras , Ácido Butírico/farmacología , Animales , Mucosa Intestinal , Intestinos , Ratones , Ratones Endogámicos C57BL
13.
Cell Physiol Biochem ; 54(1): 15-26, 2020 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-31916734

RESUMEN

BACKGROUND/AIMS: The primary cilium is a nanoscale membrane protrusion believed to act as a mechano-chemical sensor in a range of different cell types. Disruptions in its structure and signalling have been linked to a number of medical conditions, referred to as ciliopathies, but remain poorly understood due to lack of techniques capable of investigating signal transduction in cilia at nanoscale. Here we set out to use latest advances in nanopipette technology to address the question of ion channel distribution along the structure of primary cilium. METHODS: We used glass nanopipettes and Scanning Ion Conductance Microscopy (SICM) to image 3D topography of intact primary cilia in inner medullary collecting duct (IMCD) cells with nanoscale resolution. The high-resolution topographical images were then used to navigate the nanopipette along the structure of each cilium and perform spatially resolved single-channel recordings under precisely controlled mechanical and chemical stimulation. RESULTS: We have successfully obtained first single-channel recordings at specific locations of intact primary cilia. Our experiments revealed significant differences between the populations of channels present at the ciliary base, tip and within extra-ciliary regions in terms of mean conductance and sensitivity to membrane displacement as small as 100 nm. Ion channels at the base of cilium, where mechanical strain is expected to be the highest, appeared particularly sensitive to the mechanical displacement. CONCLUSION: Our results suggest the distribution of ion channels in the membrane of primary cilia is non-homogeneous. The relationship between the location and function of ciliary ion channels could be key to understanding signal transduction in primary cilia.


Asunto(s)
Membrana Celular/metabolismo , Cilios/metabolismo , Canales Iónicos/metabolismo , Nanotecnología/métodos , Potenciales de Acción/efectos de los fármacos , Adenosina Trifosfato/metabolismo , Adenosina Trifosfato/farmacología , Animales , Células Cultivadas , Células Epiteliales/citología , Células Epiteliales/metabolismo , Mecanotransducción Celular , Ratones
14.
Cancer Immunol Immunother ; 69(2): 199-213, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31982939

RESUMEN

Neutrophils play a major role in tumor biology. Among other functions, neutrophils can release extracellular traps (NETs), mesh-like structures of decondensed chromatin fibers, in a process termed NETosis. Originally characterized as an antimicrobial mechanism, NETosis has been described in cancer, but cancer-related predisposition is not clear. In the current study, we investigated the predisposition of circulating neutrophils to release NETs in lung cancer and the impact of G-CSF on this function, comparing circulating neutrophils isolated from cancer patients to the LLC and AB12 mouse models. We find that neutrophils from both healthy donors and cancer patients display high NETotic potential, with 30-60% of cells undergoing NETosis upon PMA stimulation. In contrast, neutrophils isolated from tumor-bearing mice displayed only 4-5% NETotic cells, though significantly higher than naive controls (1-2%). Despite differential mechanisms of activation described, Ionomycin and PMA mainly triggered suicidal rather than vital NETosis. G-CSF secreting tumors did not increase NETotic rates in murine neutrophils, and direct G-CSF stimulation did not promote their NET release. In contrast, human neutrophils strongly responded to G-CSF stimulation resulting also in a higher response to PMA + G-CSF stimulation. Our data show clear differences in NETotic potentials between human and murine neutrophils. We do not find a predisposition of neutrophils to release NETs in lung cancer patients compared to healthy controls, whereas cancer may modulate neutrophils' NETotic potential in mice. G-CSF secreted from tumors differentially affects murine and human NETosis in cancer. These important differences should be considered in future studies of NETosis in cancer.


Asunto(s)
Trampas Extracelulares/fisiología , Neoplasias Pulmonares/inmunología , Neutrófilos/fisiología , Animales , Línea Celular Tumoral , Trampas Extracelulares/efectos de los fármacos , Factor Estimulante de Colonias de Granulocitos/farmacología , Humanos , Ionomicina/farmacología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Acetato de Tetradecanoilforbol/farmacología
15.
Cell Physiol Biochem ; 54(1): 40-52, 2020 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-31942786

RESUMEN

BACKGROUND/AIMS: AIRE is known for its involvement in autoreactive T-cell deletion in thymic epithelium. Though extrathymic expression of AIRE is well documented, the functional relevance of AIRE in non-thymus tissues is emerging. AIRE is expressed in neonatal and adult testis, and has been implicated in sporadic germ cell apoptosis in developing testis. In this study we examined whether AIRE has any role in inducing apoptosis in cultured spermatogonial cells. METHODS: We over-expressed AIRE or CARD domain of AIRE in GC1-spg cells and evaluated its impact on cell cycle using fluorescence activated cell sorting following Hoechst 33342 staining. Apoptosis was assayed using Annexin-V staining. Caspase-3 cleavage was assessed on western blots and caspase-3 expression was quantitated using realtime PCR. RESULTS: We report that C18-4 cells which are derived from Type A spermatogonia expressed AIRE, while GC1-spg which is closer to Type B spermatogonia was negative for AIRE expression. Overexpression of AIRE or CARD domain of AIRE induced Caspase-3 expression in GC1-spg cells. Silencing of AIRE in C18-4 cells inhibited Caspase-3 expression. When overexpressed, AIRE and CARD brought about a very negligible increase in germ cell death and resulted in altered cell cycle pattern with a reduction in G1 phase. This was not associated with any increase in activation of Caspase-3. CONCLUSION: We conclude that the CARD domain of AIRE enhances caspase-3 expression through possible direct DNA binding and triggers non-apoptotic downstream signaling in cultured spermatogonial cells.


Asunto(s)
Apoptosis , Caspasa 3/genética , Espermatogonias/citología , Factores de Transcripción/genética , Animales , Línea Celular , Humanos , Masculino , Ratones , Espermatogonias/metabolismo , Testículo/citología , Testículo/metabolismo , Regulación hacia Arriba
16.
Cell Host Microbe ; 27(1): 1-3, 2020 01 08.
Artículo en Inglés | MEDLINE | ID: mdl-31951820

RESUMEN

The role of gut bacteria in complications of heart inflammation has previously not been fully understood. Gil-Cruz et al. (2019) demonstrate a commensal Bacteroides species that triggers a cross-immune response against a bacterial protein and heart epitope, causing cardiomyopathy. This study links commensal bacteria with heart inflammation mechanistically in mice and correlatively in patients.


Asunto(s)
Cardiomiopatías , Microbiota , Animales , Bacterias , Bacteroides , Humanos , Ratones , Péptidos
17.
J Biomed Nanotechnol ; 16(1): 1-13, 2020 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-31996281

RESUMEN

Targeted drug delivery systems have currently demonstrated considerable potential clinical benefits in cancer treatment. Curcumin has become a candidate anti-tumor drug for the therapy of glioblastoma multiforme (GBM) by increasing cell apoptosis and suppressing cell proliferation. In current research, we explored a novel targeted drug delivery system with a self-assembly measure by curcumin, MPEG-PLA and Fa-PEG-PLA. Compared with free curcumin and Cur/MPEG-PLA, Cur/Fa-PEG-PLA can remarkably suppress the growth of GL261 cells and promote apoptotic rate. Moreover, after the procession of tumor-bearing mice with curcumin/Fa-PEG-PLA complex, tumor growth in subcutaneous and intracranial tumor models were repressed via suppressing angiogenesis and facilitating apoptosis in vivo. The Curcumin/Fa-PEG-PLA nanoparticle may be a novel drug for the therapy of GBM.


Asunto(s)
Curcumina , Glioma , Animales , Antineoplásicos , Línea Celular Tumoral , Portadores de Fármacos , Ácido Fólico , Ratones , Micelas , Polietilenglicoles
18.
J Biomed Nanotechnol ; 16(1): 40-53, 2020 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-31996284

RESUMEN

In recent decades, a large number of research studies have been conducted to improve the treatment strategy against epithelial ovarian cancer, but women in advanced stage still have poor outcomes. The development of advanced treatments must be continued to overcome the limitation. Docetaxel, a semi-synthetic product derived from the Pacific Taxus extract, has been studied for many years for its potent anticancer applications. Aiming to solve the problems of its highly lipophilicity, insolubility and adverse side effects, nanocarriers were applied. Relying on the integration of nanoparticles which had optimized sizes, shapes, and surface properties, the effect of docetaxel was enhanced. In this study, we designed a novel drug loaded gel-forming nanoparticle system (Doc-NMs-hydrogel composites), which acted as a sustained drug depot for docetaxel. Docetaxel was encapsulated into MPEG-PCL and then into blank thermosensitive hydrogel Pluronic F-127. Characterization showed that the prepared Doc-NMs had high drug loading (7%), minor particle size (37 nm), relatively good water solubility. Moreover, the cytotoxicity, apoptosis induction and the antitumor effects of Doc-NMs-hydrogel composites on mice abdominal SKOV-3 ovarian cancer model were investigated in vivo. Compared with other groups, at the same dosage, Doc-NMs-hydrogel composites show better apoptosis induction and cell growth inhibition. In conclusion, the prepared Doc-NMs-hydrogel composites enhanced anti-tumor activity by increasing local docetaxel concentration, maintaining stable and sustained drug release, prolonging drug retention time in tumors, and reducing toxicity to normal tissues. Doc-NMs-hydrogel composites might have great potential clinical application in anti-ovarian cancer activity.


Asunto(s)
Neoplasias Ováricas , Animales , Antineoplásicos , Línea Celular Tumoral , Docetaxel , Portadores de Fármacos , Sistemas de Liberación de Medicamentos , Femenino , Humanos , Hidrogeles , Ratones , Micelas , Nanopartículas , Taxoides
19.
J Biomed Nanotechnol ; 16(1): 85-100, 2020 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-31996288

RESUMEN

Plumbagin, a natural naphthoquinone from the officinal leadwort, has recently been shown to exert promising anti-cancer effects. However, its therapeutic use is hampered by its failure to specifically reach tumors after intravenous administration, without secondary effects on normal tissues. Its poor solubility in water and rapid elimination following in vivo administration further limit its potential use. We hypothesize that the entrapment of plumbagin within PEGylated PLGA nanoparticles conjugated with transferrin, whose receptors are overexpressed on many types of cancer cells, could lead to a selective delivery of the drug to tumors following intravenous administration and enhance its chemotherapeutic effects. The objectives of this study were therefore to prepare and characterize transferrin-conjugated, PEGylated PLGA nanoparticles entrapping plumbagin, and to assess their anti-cancer efficacy in vitro as well as in tumor-bearing mice. The intravenous administration of transferrin-conjugated PEGylated PLGA nanoparticles resulted in the complete suppression of 10% of B16-F10 tumors and regression of 30% of the tumors, with improvement of the animal survival compared to controls. The treatment was well tolerated by the animals. Transferrin-bearing PEGylated PLGA nanoparticles entrapping plumbagin are therefore highly promising therapeutic systems, able to lead to tumor regression and even suppression after intravenous administration without visible toxicity.


Asunto(s)
Nanopartículas , Animales , Línea Celular Tumoral , Ratones , Naftoquinonas , Transferrina
20.
J Biomed Nanotechnol ; 16(1): 125-135, 2020 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-31996291

RESUMEN

Theranostic nanosystems encompassing imaging reagents and therapeutic genes are promising for concurrent tumor diagnosis and gene therapy. In this work, we developed bioresponsive gadolinium (Gd)-based nanopolyplexes (denoted as Gdplexes) for in vivo tumor theranostic applications. Gdplexes were generated by a hierarchical assembly method involving the neutralization of DNA with a Gd-chelated bioreducible cationic polyurethane (termed as GdCPUA), which was followed by condensation of DNA with a cationic dextran conjugate (DP800). By adjusting GdCPUA/DP800 ratios, the resultant Gdplexes had GdCPUA/DNA complex as an inner core and a dextran outer shell; thus, Gdplexes exhibit an improved colloidal stability under physiological conditions and perform active gene release in an intracellular reductive environment. In vitro tests against cancer cells revealed that optimized Gdplexes afforded comparable transfection efficiency to that of the 25 kDa branched polyethylenimine used as a positive control. Additionally, the Gdplexes could robustly transfer small hairpin RNA plasmids to silence vascular endothelial growth factor expression in SKOV-3 cells. In vivo, the Gdplexes loaded with plasmid were practical for systemic gene delivery via intravenous administration, yielding marked growth repression of an SKOV-3 tumor xenograft in a BALB/c nude mouse model. The tumor could be visualized by T1-weighted magnetic resonance (MR) imaging. Such efficient gene therapy had no adverse effects on hepatorenal functions and weight gain in the mouse. This work highlights Gdplexes as biosafe and robust nanocarriers for tumor theranostic applications in vivo.


Asunto(s)
Nanomedicina Teranóstica , Animales , Línea Celular Tumoral , Terapia Genética , Imagen por Resonancia Magnética , Ratones , Ratones Endogámicos BALB C , Factor A de Crecimiento Endotelial Vascular
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