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OBJECTIVE: Heart rate-corrected QT interval (QTc) and QTc dispersion (QTcd) are increased and associated with ventricular arrhythmia and an increase in sudden death in a variety of diseases. This study aimed to examine QTc and QTcd in pulmonary hypertension and assess their relationship with pulmonary arterial pressure. PATIENTS AND METHODS: A total of 201 patients who had undergone right heart catheterization for a preliminary diagnosis of pulmonary hypertension between December 2003 and July 2008 were included in the study. Resting 12-lead electrocardiogram was recorded. QT interval was measured manually and corrected using Bazett's formula. Patients were divided into groups with mild-to-moderate and severe pulmonary hypertension and a control group according to mean pulmonary arterial pressure. RESULTS: In all observed cases, mean QTc was higher in severe pulmonary hypertension than in controls (428.6 +/- 32.8 ms vs. 411.1 +/- 28.4 ms, P = 0.018) and QTcd was higher in mild-to-moderate (60.1 +/- 17.2 ms) and severe pulmonary hypertension (63.9 +/- 20.5 ms) than in controls (47.3 +/- 10.6 ms) (P = 0.031; P = 0.004). In men, there was no significant difference in mean QTc and QTcd. In women, mean QTc was higher in severe pulmonary hypertension than in controls (436.1 +/- 39.4 ms vs. 407.6 +/- 24.8 ms, P = 0.037) and QTcd was higher in severe pulmonary hypertension (68.5 +/- 20.9 ms) than in both the controls (45.1 +/- 12.6 ms) and patients with mild-to-moderate pulmonary hypertension (58.6 +/- 14.7 ms) (P = 0.002; P = 0.003). In addition, in women with pulmonary hypertension, mean QTc and QTcd were positively correlated to mean pulmonary arterial pressure (r = 0.207, P = 0.03; r = 0.236, P = 0.012). CONCLUSIONS: In women with pulmonary hypertension, mean QTc and QTcd are positively correlated to mean pulmonary arterial pressure and are significantly increased in patients with severe pulmonary hypertension.
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Electrocardiografía , Frecuencia Cardíaca/fisiología , Hipertensión Pulmonar/fisiopatología , Síndrome de QT Prolongado/fisiopatología , Procesamiento de Señales Asistido por Computador , Adulto , Análisis de Varianza , Índice de Masa Corporal , China , Muerte Súbita Cardíaca/etiología , Femenino , Humanos , Hipertensión Pulmonar/diagnóstico , Síndrome de QT Prolongado/diagnóstico , Masculino , Persona de Mediana Edad , Presión Esfenoidal Pulmonar/fisiología , Factores de Riesgo , Factores Sexuales , Taquicardia Ventricular/diagnóstico , Taquicardia Ventricular/fisiopatologíaRESUMEN
Epithelial-mesenchymal transformation(EMT) exists in embryonic development and is closely related to cell migration and invasion. The increased EMT level in tumors showed that E-cadherin was replaced by N-cadherin, and the expression of interstitial markers such as α-SMA and vimentin was up-regulated. It has been reported that lupeol can reduce the expression of matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9(MMP-9) and N-cadherin to inhibit the metastasis of osteoma cells. However lupeol has been less studied in liver cancer. Therefore, this paper investigated the effect of lupanol on invasion and metastasis of human hepatoma cell line HepG2 and SK-HEP-1 and its possible mechanism. MTT assay and Annexin V/PI double staining were used to investigate the effect of lupeol on activity and apoptosis of HepG2 cells and SK-HEP-1 cells. Moreover, the effect of lupeol on the invasion of HepG2 cells and SK-HEP-1 cells were evaluated by Transwell assay. The expressions of E-cadherin, N-cadherin, α-SMA, vimentin and MMP-9 were measured by Western blot. The model of subcutaneous transplantation of nude mice and the lung metastasis model of H22 hepatocellular carcinoma cells were established to evaluate the efficacy of lupeol in vivo on tumor growth and lung metastasis by HE staining combined with immunohistochemical assay. The results showed that lupeol inhibited the activity and invasion of HepG2 cells and SK-HEP-1 cells in a dose-dependent manner and induced apoptosis. Western blot showed that the expression of E-cadherin, a landmark protein for EMT, was induced by lupeol, and the expressions of N-cadherin, α-SMA, vimentin and MMP-9 were decreased. In vivo experiments showed that lupeol inhibited tumor growth in mice bearing xenograft. In addition, immunohistochemical experiments confirmed that lupeol could up-regulate the expression of E-cadherin in tumor tissues of nude mice, reduce the expression of N-cadherin, and inhibit the metastasis of liver cancer H22 cells in the lungs of mice. The above results indicated that the mechanism of lupeol inhibiting the invasion and metastasis of HCC cells may be related to the regulation of EMT process.
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Animales , Humanos , Ratones , Carcinoma Hepatocelular , Línea Celular Tumoral , Movimiento Celular , Transición Epitelial-Mesenquimal , Regulación Neoplásica de la Expresión Génica , Células Hep G2 , Neoplasias Hepáticas , Metaloproteinasa 2 de la Matriz/metabolismo , Ratones Desnudos , Invasividad Neoplásica , Triterpenos PentacíclicosRESUMEN
Based on metabolomics,the metabolites of larvae zebrafish with overdose of Panax notoginseng saponins( PNS) were compared with those in normal group of larvae zebrafish to investigate the possible toxicity mechanism of overdose PNS in larvae zebrafish. An experimental animal model of long-term toxicity induced by PNS overdose was established by administering 1-6 dpf at low,medium and high doses of PNS,respectively. The ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry( UPLC-Q-TOF-MS) technique was combined with principal component analysis( PCA) and orthogonal partial least squares discriminant analysis( OPLS-DA) to screen and identify biomarkers associated with toxicity,and then the MetaboAnalyst database was used to analyze metabolism-related pathways. The results showed that the metabolites of each group could be distinguished distinctly,and they deviated more from the normal group in a time and dose dependent manner. Twenty-nine potential biomarkers related to toxicity( VIP>1,P<0. 05) were identified preliminarily,mainly involving six metabolic pathways. From the metabonomics point of view,the toxicity mechanism of overdose PNS may be related to the disorders of lipid metabolism,amino acid metabolism and energy metabolism.
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Animales , Aminoácidos , Metabolismo , Cromatografía Líquida de Alta Presión , Metabolismo Energético , Larva , Metabolismo de los Lípidos , Espectrometría de Masas , Metabolómica , Panax notoginseng , Toxicidad , Saponinas , Toxicidad , Pruebas de Toxicidad Aguda , Pez CebraRESUMEN
Metabolic transformation in vivo is a critical approach in the study of toxicity, but real-time dynamic observation of the transformation is difficult. We proposed that zebrafish toxicity/metabolism synchronization may be used in the analysis of toxicity of Folium Epimedium (Yinyanghuo for Chinese, YYH) and the toxicity may be reduced by Radix Morindae Officinalis (Bajitian for Chinese, BJT). Healthy zebrafish embryos 1 day post fertilization (1 dpf) were exposed to different concentrations of YYH, total flavonoids of YYH (YTF), representative flavonoids (epimedin C and icariin) and their respective in combination with BJT. Death numbers of the embryos or larvals were counted during 1-5 days after dosing (2-6 dpf); embryonic micro-morphology of zebrafish (3 dpf) was observed and pictures were taken. The blank vehicle (0.4% DMSO) was used in the control group, and LC50 value of 2 to 6 dpf was calculated by SPSS16.0. A relative safe concentration was sampled every day to analyze the dynamic metabolites changes of major flavonoids of YYH. The results showed that epimedin A/B/C (EA/EB/EC) and icariin, the major flavonoids of YYH, were dynamically transformed into major metabolites of sagittatoside C (SC) and baohuoside I (BI) by zebrafish. BI was mainly derived from EA, EB and icariin. Neither original form nor their metabolite BI can cause zebrafish poisoning. SC was mainly derived from EC, and its accumulation was closely related to the toxicity of YYH, YTF and EC. After combination with BJT, the metabolism of EC was slowed down and the toxicity was alleviated. Zebrafish toxicity/metabolism synchronization revealed that the toxicity of EC of YYH was increased after metabolism into SC, which maybe the key potential poisonous factor of YYH, and BJT can reduce the toxicity by slowing down the metabolism rate of EC. The data provides new ideas and methods in the study of toxic substances in Chinese medicine and mechanism of detoxicity by combination.
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The increasingly apparent liver injury problems of bone strengthening Chinese medicines have brought challenges for clinical application, and it is necessary to consider both effectiveness and safety in screening anti-osteoporosis Chinese medicines. Metabolic transformation is closely related to drug efficacy and toxicity, so it is significant to comprehensively consider metabolism-action/toxicity(M-Act/Tox) for screening anti-osteoporosis Chinese medicines. The current evaluation models and the number of compounds(including metabolites) severely restrict efficient screening in vivo. By referring to previous relevant research and domestic and abroad literature, zebrafish M-Act/Tox integrative method was put forward for efficiently screening anti-osteoporosis herb medicines, which has organically integrated zebrafish metabolism model, osteoporosis model and toxicity evaluation method. This method can break through the bottleneck and blind spots that trace compositions can't achieve efficient and integrated in vivo evaluation, and realize both efficient and comprehensive screening on anti-osteoporosis traditional medicines based on in vivo process taking both safety and effectiveness into account, which is significant to accelerate discovery of effective and safe innovative traditional Chinese medicines for osteoporosis.
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The safety of traditional Chinese medicine (TCM) has received the widespread attention in recent years. Hepatotoxicity of TCM is one of the key problems of the safety of TCM. This article summarized research progress and application prospect in the mechanism of TCM hepatotoxicity, biomarkers, toxic omics database, prevention of hepatotoxicity of the liver cell lines, subcellular fraction, three-dimensional cultivation models, the model animals, aiming to provide theoretical basis for TCM toxicity evaluation and technical guidelines, thus promoting the development of TCM toxicity studies. Hope for Chinese medicine liver toxicity evaluation method provides the theoretical foundation and technical guidelines, promote the development and improvement of TCM liver toxicity research system.
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This article firstly established a new efficient method for screening anti-osteoporosis ingredients, which used two-dimensional zebrafish model combined with hyphenated chromatographic techniques to evaluate anti-osteoporosis activities of epimedin A and its metabolite baohuoside I. Adult zebrafish was used for metabolism of epimedin A in 0.5% DMSO, and LC-MS was used for analysis of the metabolite, which was captured by HPLC, and prednisolone-induced osteoporosis model of zebrafish was used to evaluate the anti-osteoporotic activities of trace amounts of epimedin A and baohuoside I. The results indicated that epimedin A and baohuoside I can prevent prednisolone-induced osteoporosis in zebrafish. The developed method in this paper enables the separation, enrichment and analysis of micro-amount metabolite of epimedin A, and anti-osteoporosis activities in vivo of epimedin A and baohuoside I was simple and efficient screening resorting to zebrafish osteoporosis mode. This paper would provide new ideas and methods for a rapid and early discovery of anti-osteoporosis activities of micro-ingredients and its metabolite of traditional Chinese medicine.
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Animales , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Evaluación Preclínica de Medicamentos , Medicamentos Herbarios Chinos , Farmacología , Flavonoides , Metabolismo , Farmacología , Espectrometría de Masas , Osteoporosis , Quimioterapia , Pez CebraRESUMEN
<p><b>OBJECTIVE</b>To break through the restrictions of the evaluation model and the quantity of compounds by using the two-dimensional zebrafish model combined with chromatographic techniques, and establish a new method for the high-throughput screening of active anti-osteoporosis components.</p><p><b>METHOD</b>According to the research group-related studies and relevant foreign literatures, on the basis of the fact that the zebrafish osteoporosis model could efficiently evaluate the activity, the zebrafish metabolism model could efficiently enrich metabolites and the chromatographic techniques could efficiently separate and analyze components of traditional Chinese medicines, we proposed that the inherent combination of the three methods is expected to efficiently decode in vivo and in vitro efficacious anti-osteoporosis materials of traditional Chinese medicines.</p><p><b>RESULT AND CONCLUSION</b>The method makes it simple and efficient in the enrichment, separation and analysis on components of traditional Chinese medicines, particularly micro-components and metabolites and the screening anti-osteoporosis activity, fully reflects that efficacious materials of traditional Chinese medicines contain original components and metabolites, with characteristic of "multi-components, multi-targets and integral effect", which provides new ideas and methods for the early and rapid discovery of active anti-osteoporosis components of traditional Chinese medicines.</p>
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Animales , Humanos , Cromatografía , Métodos , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Métodos , Medicamentos Herbarios Chinos , Usos Terapéuticos , Medicina Tradicional China , Métodos , Osteoporosis , Quimioterapia , Fitoterapia , Métodos , Reproducibilidad de los Resultados , Pez Cebra , FisiologíaRESUMEN
Objective: To investigate the in vivo metabolism of the combination of tanshinone IIA (Tan IIA), cryptotanshinone (Cry), and tanshinone I (Tan I) in zebrafish and discuss the possibility and rationality of using zebrafish in multi-component metabolism of Chinese materia medica (CMM). Methods: The zebrafish was exposed to the 1% DMSO solution of the combinations with Tan IIA, Cry, and Tan I. High performance liquid chromatography coupled with ion-trap mass spectrometry (HPLC-MS) method was used to calculate the relative molecular weight of the metabolites based on the excimer ion peak. The metabolites in solution and zebrafish after the combination being exposed to zebrafish for 24 h were identified through comparing with the literature data and reference substances. Results: After the combination being exposed to zebrafish, hydroxylation and dehydrogenation products of Tan IIA and Cry were obtained while the metabolites of Tan I was not found. Conclusion: The metabolic mechanism of zebrafish against the combination highly consists with those in rats or rat liver microsomes, which indicates that the metabolism of zebrafish against the combination is rational with the advantages of less amount of compound, lower cost, simpler operation, and higher efficiency. The above results provide the reference for using zebrafish in the metabolic study on complex CMM system.
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OBJECTIVE: To study the metabolism of chrysin by model organism zebrafish for the first time, thus to investigate the reasonability of applying zebrafish in drug phase II metabolism. METHODS: Zebrafish was exposed to chrysin solution for 24 h, and the samples of solution and zebrafish body were analyzed by high performance liquid chromatography-electrospray mass spectrometry (HPLC-ESI-MS) method. The quasi-molecular ions of compounds in both negative and positive mode were observed for molecule mass information, and the potential structures of the compounds were identified by studying on the mass spectra and comparing with reference data or standards. RESULTS: In addition to the parent compound, three phase II metabolites were identified, including two glucuroni-dation products and one sulfation product. CONCLUSION: The metabolism of chrysin in zebrafish is highly consistent with that identified by current in vivo and in vitro methods, which indicates that it is reasonable using zebrafish to sutdy phase II metabolism. Zebrafish model has the advantages of using far less amount of compound, lower cost, higher efficiency and simple procedure, which may become a novel organism model for quick predication of metabolism of compounds and enrich the available models greatly.
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Model organism zebrafish was used to study metabolism of asperosaponin VI from Dipsacus asper Wall. ex Henry for the first time. Metabolic components of asperosaponin VI after exposing to zebrafish for 24 h were identified by high performance liquid chromatography--electrospray mass spectrometry (HPLC-ESI-MS), the separation was performed with a Zorbax C18 column using a binary gradient elution of 0.05% formic acetonitrile--0.05% formic acid water. The quasi-molecular ions of compounds in both negative and positive mode were observed for molecule mass information, and the potential structures were identified by attentive study on the deglycosylation metabolites and one hydroxylation metabolite of asperosaponin VI. The results were highly in consistent with metabolism of asperosaponin VI in rat. It can be concluded that zebrafish model can wonderfully imitate current metabolic model with advantages of small amount of lower cost, far less amount compound, higher efficiency and more simple, and can reflect integrated metabolism results of in vivo method. Zebrafish metabolic model may become a novel organism model for quick predication on metabolism of even mircoamount compound, which can enrich the available models greatly.
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Animales , Femenino , Masculino , Ratas , Cromatografía Líquida de Alta Presión , Dipsacaceae , Química , Modelos Animales , Plantas Medicinales , Química , Distribución Aleatoria , Saponinas , Metabolismo , Espectrometría de Masa por Ionización de Electrospray , Pez Cebra , MetabolismoRESUMEN
Zebrafish was selected as model animal, and glucocorticoid dexamethasone was used as a model compound to establish a rapid and high efficient osteopenia model. Zebrafish larvae at 4 days post fertilization (dpf) were exposed to a serial concentrations of dexamethasone solutions, and 0.5% DMSO was selected as the vehicle control group. All groups were incubated in 24-well plates (28.5 degrees C) until 9 dpf. In addition, effects of 10 micromol x L(-1) dexamethasone on preventing against osteopenia induced by etidronate disodium were also investigated. Zebrafish bones at 9 dpf were stained with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of bone mineralization. Results showed that dexamethasone group at 2.5, 10 and 25 micromol x L(-1) can decrease the staining area and the staining optical density values of zebrafish head bones when compared with the vehicle control group (0.5% DMSO), which suggested that dexamethasone can significantly reduce the zebrafish mineralized bone and the bone mineral density. Results also showed that 15 and 30 microg x mL(-1) etidronate disodium can increase the mineralized matrix of zebrafish head bone and prevent against osteopenia induced by dexamethasone. In conclusion, the study indicated that zebrafish can be an idea osteopenia model induced by dexamethasone.
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Animales , Densidad Ósea , Conservadores de la Densidad Ósea , Farmacología , Usos Terapéuticos , Enfermedades Óseas Metabólicas , Calcificación Fisiológica , Dexametasona , Toxicidad , Modelos Animales de Enfermedad , Ácido Etidrónico , Farmacología , Usos Terapéuticos , Larva , Pez CebraRESUMEN
<p><b>OBJECTIVE</b>To investigate the efficacy of transplantation of mesenchymal stem cells (MSC) with gelatin microspheres containing vascular endothelial growth factor in ischemic regions in infracted swine hearts.</p><p><b>METHODS</b>Twelve Chinese mini swines with infarction were randomized to receive autogenetic MSC injection to the peri-infarction area of left ventricular wall (MSC group, n = 6) or MSC transplantation with gelatin hydrogel microspheres incorporating vascular endothelial growth factor (VEGF-MSC group, n = 6). Three weeks later, left ventricular function was assessed by magnetic resonance imaging (MRI). The contrast of the MSC hypointense lesion was determined using the difference in signal intensity between the hypointense and normal myocardium divided by signal intensity of the normal region. Myocardial capillary density, the number of DAPI positive MSC and the apoptotic MSC were also determined.</p><p><b>RESULTS</b>The diameter of the microspheres averaged (104.0 +/- 22.6) microm. At 24 hours after transplantation, MSC were identified by MRI as large intramyocardial signal voids at injection sites which persisted up to 3 weeks. There was no significant difference in the contrast of the lesions and in the size of the lesions at 24 hours between two groups. At 3 weeks after injection, the size of the lesions and the contrast of the lesion were decreased (P < 0.05) in both groups. The capillary density of the injection site was significantly more in the MSC-VEGF microsphere group than that in MSC group [(15.2 +/- 5.4)/HPF vs. (10.2 +/- 5.0)/HPF, t = 2.43, P < 0.05], and there were more dense DAPI labeled MSC per high power fields in injection sites of MSC-VEGF microsphere group than that in MSC group [(354 +/- 83)/HPF vs. (278 +/- 97)/HPF, t = 3.14, P < 0.05]. Moreover, the apoptosis rate of MSCs of MSCs-VEGF microsphere group was less than that of MSC group [(6.4 +/- 4.1)% vs. (11.9 +/- 4.8)%, t = 2.97, P < 0.05].</p><p><b>CONCLUSIONS</b>MSC transplantation with gelatin hydrogel microspheres incorporating VEGF enhanced the efficacy of MSC in this swine model of myocardial infarction. MRI tracking of MSC is feasible and represents a preferred method for studying the engraftment of MSCs in infracted tissue.</p>
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Animales , Células de la Médula Ósea , Metabolismo , Gelatina , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas , Metabolismo , Microesferas , Infarto del Miocardio , Porcinos , Factor A de Crecimiento Endotelial Vascular , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To investigate whether cardiac tissue extracts from rats could mimic the cardiac microenvironment and act as a natural inducer in promoting the differentiation of bone marrow stromal cells (BMSCs) into cardiomyocytes.</p><p><b>METHODS</b>Three kinds of tissue extract or cell lysate [infarcted myocardial tissue extract (IMTE), normal myocardial tissue extract (NMTE) and cultured neonatal myocardial lysate (NML)] were employed to induce BMSCs into cardiomyocyte-like cells. The cells were harvested at each time point for reverse transcription-polymerase chain reaction (RT-PCR) detection, immunocytochemical analysis, and transmission electron microscopy.</p><p><b>RESULTS</b>After a 7-day induction, BMSCs were enlarged and polygonal in morphology. Myofilaments, striated sarcomeres, Z-lines, and more mitochondia were observed under transmission electron microscope. Elevated expression levels of cardiac-specific genes and proteins were also confirmed by RT-PCR and immunocytochemistry. Moreover, IMTE showed a greater capacity of differentiating BMSCs into cardiomyocyte-like cells.</p><p><b>CONCLUSIONS</b>Cardiac tissue extracts, especially IMTE, can effectively differentiate BMSCs into cardiomyocyte-like cells.</p>
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Animales , Masculino , Ratas , Secuencia de Bases , Células de la Médula Ósea , Biología Celular , Diferenciación Celular , Cartilla de ADN , Inmunohistoquímica , Microscopía Electrónica de Transmisión , Infarto del Miocardio , Metabolismo , Miocardio , Biología Celular , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células del Estroma , Biología CelularRESUMEN
Objective:To sieve molecular biomarkers associated with heart failure derived from arrhythmogenic right ventricular cardiomyopathy (ARVC).Methods:The comparative gene microarray analysis using individual left ventricular myocardial samples from 5 patients with heart failure resulting from ARVC undergoing transplantation and 5 matched samples from normal adult heart were performed.The accuracy rate of the differentially expressed genes obtained by gene microarray was further verified by quantitative real time RT-PCR.Results:83 genes (from a total of 35000) that were differentially expressed in diseased hearts versus normal hearts were identified.Among them thirty-seven genes were up-regulated and forty-eight genes were down-regulated in ARVC hearts compared with the normal hearts.Changes of gene expressions were most prominently observed in those belonging to the "metabolism" category.Eighty percent of the selected 30 differentially expressed genes from microarray analysis were confirmed by quantitative real time RT-PCR.The highly expressed level of atrial natriuratic peptide (ANP) in ARVC hearts that was confirmed by quantitative real time RT-PCR and immunohistochemistry was reported.Conclusion:For the first time to our knowledge,the altered expressed genes in ARVC hearts compared with the matched normal hearts were identified.The results are the base to further study the molecular mechanism and identify diseased-specific molecular biomarkers in heart failure derived from ARVC.
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<p><b>OBJECTIVE</b>To investigate whether injectable engineering heart tissue (EHT) can survive and improve heart function after transplantation into infarct area.</p><p><b>METHODS</b>Ventricular cardiomyocytes from 1-3 day-old Sprague-Dawley (SD) rats were isolated by using trypsin method, and then labeled and cultured. The left coronary of female SD rats was ligated to create a myocardial infarct model. Three weeks later, the qualified animals were randomized into four groups: EHT group (n = 12), which were transplanted with both cardiomyocytes and matrix; cell transplantation group (n = 12); matrix group (n = 12), control (n = 11). Four weeks after implantation, echocardiography and Langendorff model were used to assess heart function, and then the hearts were harvested for pathological examination. Meanwhile polymerase chain reaction (PCR) was performed to detect SRY gene on Y chromosome.</p><p><b>RESULTS</b>The grafted cells were identified in both EHT and cell transplantation group by either pathology or PCR. But in EHT group, transplanted cells formed more condensed tissue, and produced definite connected protein. Data of fraction shortness from echocardiography are showed as follows: EHT group, (22.82 +/- 3.44)%; cell transplantation group, (20.55 +/- 4.11)%, matrix group, (17.05 +/- 4.57)%; control, (19.80 +/- 3.98)% (P = 0.012). Langendorff examination revealed significant differences among four groups when left ventricular balloon volume was at the level of 0.06 ml, 0.08 ml and 0.10 ml, in which EHT group had the highest developed pressure and dp/dt.</p><p><b>CONCLUSION</b>It is feasible to fabricate injectable EHT in vitro. The fabricated EHT could survive in the myocardial infarct area after transplantation in a rat model and improve heart function due to better histological configuration.</p>
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Animales , Femenino , Ratas , Trasplante de Células , Métodos , Ecocardiografía , Infarto del Miocardio , Cirugía General , Miocitos Cardíacos , Trasplante , Ratas Sprague-Dawley , Ingeniería de TejidosRESUMEN
<p><b>OBJECTIVE</b>To investigate the feasibility of gene transfection of bone marrow mesenchymal stem cell (BMSC) by cationic lipid.</p><p><b>METHODS</b>The relative optimal transfection condition was determined by scale transfection experiment in vitro and the transient transfection efficiency of enhanced green fluorescence protein (EGFP) gene for rat BMSC was determined with Lipofectamine2000 (LP2000). The relationship between cell cycle status and the expression of the gene was analyzed. The intensity and the ratio of EGFP gene expression versus time was determined by flow cytometry. In the in vivo study, the transgenic rat BMSC was injected into the myocardium of inbred rats, and the in vivo transcription of EGFP gene and the EGFP-expressing BMSC were traced in the myocardium after transplantation using reverse transcription-polymerase chain reaction and fluorescent microscopy, respectively.</p><p><b>RESULTS</b>EGFP gene transfection efficiency in BMSC was different under different transfection condition (DNA concentration and DNA: LP2000). Cationic lipid-mediated transfection demonstrated marked toxicity to BMSC. Cell cycle status restricted the expression efficiency of the gene introduced by cationic lipid. The EGFP expressing-BMSC and in vivo transcription of the EGFP gene could be detected in rat myocardium post implantation.</p><p><b>CONCLUSION</b>Cationic lipid is an effective carrier for gene-modified cell transplantation therapy.</p>