Network Pharmacology and Experimental Validation to Explore Mechanism of Tetrahydropalmatine on Acute Myocardial Ischemia / 中国结合医学杂志

OBJECTIVE@#To explore the potential molecular mechanism of tetrahydropalmatine (THP) on acute myocardial ischemia (AMI).@*METHODS@#First, the target genes of THP and AMI were collected from SymMap Database, Traditional Chinese Medicine Database and Analysis Platform, and Swiss Target Prediction, respectively. Then, the overlapping target genes between THP and AMI were evaluated for Grene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and protein-protein interaction network analysis. The binding affinity between the protein and THP was assessed by molecular docking. Finally, the protective effects of THP on AMI model and oxygen and glucose deprivation (OGD) model of H9C2 cardiomyocyte were explored and the expression levels of target genes were detected by RT-qPCR in vivo and in vitro.@*RESULTS@#MMP9, PPARG, PTGS2, SLC6A4, ESR1, JAK2, GSK3B, NOS2 and AR were recognized as hub genes. The KEGG enrichment analysis results revealed that the potential target genes of THP were involved in the regulation of PPAR and hormone pathways. THP improved the cardiac function, as well as alleviated myocardial cell damage. Furthermore, THP significantly decreased the RNA expression levels of MMP9, PTGS2, SLC6A4, GSK3B and ESR1 (P<0.05, P<0.01) after AMI. In vitro, THP significantly increased H9C2 cardiomyocyte viability (P<0.05, P<0.01) and inhibited the RNA expression levels of PPARG, ESR1 and AR (P<0.05, P<0.01) in OGD model.@*CONCLUSIONS@#THP could improve cardiac function and alleviate myocardial injury in AMI. The underlying mechanism may be inhibition of inflammation, the improvement of energy metabolism and the regulation of hormones.

Association between genetic predisposition to childhood obesity and the risk of adult ischemic heart disease in China / 中华流行病学杂志

Objective: To examine the associations of childhood obesity, assessed by genetic variations of childhood body mass index (BMI), with the risk of adult ischemic heart disease (IHD) and major coronary event (MCE). Methods: More than 69 000 participants from the China Kadoorie Biobank were genotyped. After excluding those with coronary heart disease, stroke, or cancer at baseline, a total of 64 454 participants were included in this study. Based on genome-wide significant single nucleotide polymorphisms (SNPs), childhood BMI genetic risk score were constructed for every participant and divided into quintiles, with the lowest quintile as the low genetic risk group and the highest quintile as the high genetic risk group. Cox proportional hazards regression models were used to estimate the association between genetic predisposition to childhood obesity and the risk of ischemic heart disease. Results: During a median of 10.7 years of follow-up, 7 073 incident cases of IHD and 1 845 cases of MCE were documented. After adjusting for sex, age, region, and the first ten genetic principal components, the HRs (95%CIs) for IHD and MCE in the high genetic risk group were 1.10 (1.02-1.18) and 1.10 (0.95-1.27), compared with the low genetic risk group. IHD risk increased by 4% (2%-6%) for each one standard deviation increase in genetic risk score (trend P=0.001). After further adjustment for baseline BMI, the differences between genetic risk groups were not statistically significant, but there was still a linear trend between genetic risk score and IHD risk (trend P=0.019). Conclusions: IHD risk increased with genetic predisposition to childhood obesity, suggesting that childhood obesity is an important risk factor for the development of IHD in China. As an easily identifiable feature, changes of childhood BMI should be monitored regularly to realize early intervention of IHD in adults.

Molecular mechanism of Qishen Yiqi Dripping Pills in treating myocardial ischemia:a study based on HIF-1 signaling pathway / 中国中药杂志

Qishen Yiqi Dripping Pills(QSYQ) are used clinically to treat various myocardial ischemic diseases, such as angina pectoris, myocardial infarction, and heart failure; however, the molecular mechanism of QSYQ remains unclear, and the scientific connotation of traditional Chinese medicine(TCM) compatibility has not been systematically explained. The present study attempted to screen the critical pathway of QSYQ in the treatment of myocardial ischemia by network pharmacology and verify the therapeutic efficacy with the oxygen-glucose deprivation(OGD) model, in order to reveal the molecular mechanism of QSYQ based on the critical pathway. The key targets of QSYQ were determined by active ingredient identification and target prediction, and underwent pathway enrichment analysis and functional annotation with David database to reveal the biological role and the critical pathway of QSYQ. Cell counting Kit-8(CCK-8), lactate dehydrogenase(LDH), and Western blot tests were launched on high-content active ingredients with OGD cell model to reveal the molecular mechanism of QSYQ based on the critical pathway. The results of network pharmacology indicated that QSYQ, containing 18 active ingredients and 82 key targets, could protect cardiomyocytes by regulating biological functions, such as nitric oxide biosynthesis, apoptosis, inflammation, and angiogenesis, through TNF signaling pathway, HIF-1 signaling pathway, PI3 K-Akt signaling pathway, etc. HIF-1 signaling pathway was the critical pathway. As revealed by CCK-8 and LDH tests, astragaloside Ⅳ, salvianic acid A, and ginsenoside Rg_1 in QSYQ could enhance cell viability and reduce LDH in the cell supernatant in a concentration-dependent manner(P<0.05). As demonstrated by the Western blot test, astragaloside Ⅳ significantly down-regulated the protein expression of serine/threonine-protein kinase(Akt1) and hypoxia-inducible factor 1α(HIF-1α) in the HIF-1 signaling pathway, and up-regulated the protein expression of vascular endothelial growth factor A(VEGFA). Salvianic acid A significantly down-regulated the protein expression of upstream phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3 CA) and downstream HIF-1α of Akt1. Ginsenoside Rg_1 significantly down-regulated the expression of HIF-1α protein and up-regulated the expression of VEGFA. The therapeutic efficacy of QSYQ on myocardial ischemia was achieved by multiple targets and multiple pathways, with the HIF-1 signaling pathway serving as the critical one. The active ingredients of QSYQ could protect cardiomyocytes synergistically by regulating the targets in the HIF-1 signaling pathway to inhibit its expression.

Mechanism of paeonol combined with paeoniflorin against myocardial ischemia injury:based on proteomics / 中国中药杂志

The study aims to investigate the effect of the compatibility of paeonol and paeoniflorin(hereinafter referred to as the compatibility) on the expression of myocardial proteins in rats with myocardial ischemia injury and explore the underlying mechanism of the compatibility against myocardial ischemia injury. First, the acute myocardial infarction rat model was established by ligation of the anterior descending branch of the left coronary artery. The model rats were given(ig) paeonol and paeoniflorin. Then protein samples were collected from rat cardiac tissue and quantified by tandem mass tags(TMT) to explore the differential proteins after drug intervention. The experimental results showed that differential proteins mainly involved phagocytosis engulfment, extracellular space, and antigen binding, as well as Kyoto encyclopedia of genes and genomes(KEGG) pathways of complement and coagulation cascades, syste-mic lupus erythematosus, and ribosome. In this study, the target proteins and related signaling pathways identified by differential proteomics may be the biological basis of the compatibility against myocardial ischemia injury in rats.

Caracterización de los factores de riesgo en pacientes con cardiopatía isquémica Hospital "Mártires del 9 de Abril", período 2016-2017 / Characterization of risk factors in patients with ischemic heart disease. Hospital "Martires del 9 de abril", period 2016-2017

RESUMEN Introducción: las enfermedades cardiovasculares constituyen la principal causa de muerte en la mayoría de los países. Se describen los factores de riesgo para enfermedad coronaria como no modificables: edad, sexo y antecedentes familiares; y modificables relacionados al estilo de vida: tabaquismo, dislipidemia, obesidad, sedentarismo, diabetes, uso abusivo de alcohol y la enfermedad hipertensiva. Objetivo: caracterizar los factores de riesgo asociados a la cardiopatía isquémica en Atención Secundaria de Salud. Materiales y métodos: estudio observacional, descriptivo transversal en pacientes ingresados en el Hospital "Mártires del 9 de Abril" de Sagua la Grande, en el periodo comprendido entre los años 2016 y 2017. Integraron la muestra 96 pacientes que ingresaron con diagnóstico de cardiopatía isquémica. Se describieron las características demográficas de los mismos; fueron identificados los factores de riesgo y se determinó la frecuencia de asociación de otras formas clínicas de ateromatosis. Resultados: los pacientes fueron mayores de 60 años de edad; la mayoría tenían color de la piel blanca; presentaban antecedentes patológicos familiares de cardiopatía isquémica; las principales formas de cardiopatía isquémica fueron: angina e insuficiencia cardiaca; todos los pacientes presentaron uno o más factores de riesgo cardiovascular, los más significativos fueron, hipertensión arterial, tabaquismo aumento de la circunferencia abdominal y personalidad tipo "A". Conclusiones: la mayoría de los pacientes exhibieron alteraciones en el electrocardiograma: descenso del segmento ST, bloqueo de rama izquierda del haz de His y fibrilación auricular; se observó hipertrigliceridemia y se apreció asociación entre enfermedad renal crónica y angina.

ABSTRACT Introduction: cardiovascular diseases are the main cause of death in most of the countries. The risk factors for coronary disease are described as unmodifiable: age, sex and family history; and modifiable related to lifestyle: smoking, dyslipidemia, obesity, sedentary lifestyle, diabetes, abusive use of alcohol and hypertensive disease. Objective: to characterize the risk factors associated to ischemic heart disease in secondary health care. Material and methods: observational, cross-sectional, descriptive study in patients admitted in "Mártires del 9 de Abril" Hospital, Sagua la Grande, between 2016 and 2017. The sample consisted of 96 patients admitted with a diagnosis of ischemic heart disease. Their demographic characteristics were described; the risk factors were identified and the frequency of association of other clinical forms of atheromatosis was determined. Results: the patients were aged more than 60 years; most of them were white people and had family pathological antecedents of ischemic heart disease; the main forms of ischemic heart disease were angina and heart failure; all patients showed one or more cardiovascular risk factors being arterial hypertension, smoking, increase of abdominal circumference and type A personality the most significant ones. Conclusions: the majority of patients showed alterations in the electrocardiogram: ST segment decrease, His bundle left branch blockage and atrial fibrillation; hypertriglyceridemia was observed and there was an association between chronic kidney disease and angina.

Diabetes mellitus tipo 2 y cardiopatía isquémica: fisiopatología, regulación génica y futuras opciones terapéuticas / Type II diabetes and ischemic heart disease: pathophysiology, gen regulation and future therapeutic options

Resumen: En los últimos años, la diabetes mellitus tipo 2 (DM2) ha evolucionado en forma epidémica, experimentando un rápido crecimiento y afectando a millones de individuos a nivel mundial. La cardiopatía isquémica es la principal causa de mortalidad en los pacientes diabéticos, quienes poseen un mayor riesgo cardiovascular respecto a los no diabéticos. La DM2 y la cardiopatía isquémica se caracterizan por ser prevenibles, sin embargo, existen diversos factores de riesgo comunes que contribuyen a su desarrollo. Los mecanismos que explican la ateroesclerosis acelerada y el incremento de riesgo de enfermedades cardiovasculares en los pacientes diabéticos tipo 2 incluyen a la hiperglicemia, dislipidemia y la inflamación del endotelio vascular. La diabetes es resultado de una interacción compleja entre la genética y el medio ambiente. Recientemente se han descrito varios genes implicados en el desarrollo de la diabetes y cardiopatía isquémica y que podrían significar nuevas opciones terapéuticas. En este artículo se revisa la relación entre ambas patologías, los mecanismos moleculares y el descubrimiento de factores de riesgo genéticos comunes y su implicancia en el desarrollo de nuevos blancos terapéuticos.

Abstracts: In recent years, type 2 diabetes mellitus has evolved as a rapidly increasing epidemic and affects millions of people worldwide. Ischemic heart disease (IHD) is the main cause of death among diabetic patients, who have a higher cardiovascular risk than non-diabetics. Both, DM2 and IHD are characterized by being preventable, however there are several common risk factors that contribute to their development. The mechanisms that explain accelerated atherosclerosis and increased risk of cardiovascular diseases in patients with type 2 diabetes mellitus include damage by hyperglycemia, dyslipidemia and inflammation on vascular endothelium. Diabetes is the result of a complex interaction between genetics and the environment, recently, several genes have been identified that appear to be involved in diabetes and ischemic heart disease that could explain its relationship and serve as new therapeutic possibilities. In this article, we review the relationship between diabetes and ischemic heart disease, the molecular mechanisms and the discovery of genetic risk factors common to both diseases and their implication in the development of new therapeutic targets.

Long non-coding RNA-ROR aggravates myocardial ischemia/reperfusion injury

Long non-coding RNAs (lncRNAs) play an important role in the pathogenesis of cardiovascular diseases, especially in myocardial infarction and ischemia/reperfusion (I/R). However, the underlying molecular mechanism remains unclear. In this study, we determined the role and the possible underlying molecular mechanism of lncRNA-ROR in myocardial I/R injury. H9c2 cells and human cardiomyocytes (HCM) were subjected to either hypoxia/reoxygenation (H/R), I/R or normal conditions (normoxia). The expression levels of lncRNA-ROR were detected in serum of myocardial I/R injury patients, H9c2 cells, and HCM by qRT-PCR. Then, levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) were measured by kits. Cell viability, apoptosis, apoptosis-associated factors, and p38/MAPK pathway were examined by MTT, flow cytometry, and western blot assays. Furthermore, reactive oxygen species (ROS) production was determined by H2DCF-DA and MitoSOX Red probes with flow cytometry. NADPH oxidase activity and NOX2 protein levels were measured by lucigenin chemiluminescence and western blot. Results showed that lncRNA-ROR expression was increased in I/R patients and in H/R treatment of H9c2 cells and HCM. Moreover, lncRNA-ROR significantly promoted H/R-induced myocardial injury via stimulating release of LDH, MDA, SOD, and GSH-PX. Furthermore, lncRNA-ROR decreased cell viability, increased apoptosis, and regulated expression of apoptosis-associated factors. Additionally, lncRNA-ROR increased phosphorylation of p38 and ERK1/2 expression and inhibition of p38/MAPK, and rescued lncRNA-ROR-induced cell injury in H9c2 cells and HCM. ROS production, NADPH oxidase activity, and NOX2 protein levels were promoted by lncRNA-ROR. These data suggested that lncRNA-ROR acted as a therapeutic agent for the treatment of myocardial I/R injury.

Identificación y expresión diferencial del factor de transcripción asociado a microftalmia en corazón y cardiomiocitos aislados de cobayo: su posible papel en la hipertrofia y la viabilidad / Identification and differential expression of the microphthalmia-associated transcription factor in heart and isolated cardiomyocytes from Guinea pigs: Possible role in hypertrophy and viability

Introducción. El factor de transcripción asociado a la microftalmia ( Microphtalmia-Associated Transcription Factor , MITF) regula la expresión de genes específicos, pero no se conoce su expresión y su función a nivel cardiaco. Objetivos. Identificar la expresión del MITF en corazón y en cardiomiocitos aislados de cobayo, describir los cambios morfológicos asociados con su disminución y evaluar los niveles relativos de su expresión en cardiomiocitos aislados en condiciones de preacondicionamiento isquémico. Materiales y métodos. El análisis de la expresión relativa de la isoforma específica de tejido cardiaco ( heart-type MITF, MITF-H), se determinó mediante reacción en cadena de la polimerasa (PCR) en tiempo real semicuantitativa, secuenciación y Western blot . La disminución del ARNm del MITF se indujo con un ARN pequeño de interferencia ( short hairpin RNA interference , shRNAi) específico. El tamaño, el diámetro y el número de fibras musculares se evaluaron por observación directa con microscopía de luz. Resultados. Se amplificó un fragmento de 281 pb de ADNc; el análisis de la secuencia confirmó la identidad del exón 1 y la isoforma H del MITF. La interferencia del ARNm del MITF se asoció con un mayor índice cardiaco (peso corazón/peso corporal: 5,46 x 10 -3 Vs. 4,6 x 10 -3 ) y un incremento del diámetro de las fibras cardiacas (50,2±16 µm Vs. 38,7±14,7 µm; p<0,05, n=150). En los cardiomiocitos aislados en condiciones de preacondicionamiento isquémico, se observó una expresión relativa del MITF-H mayor que en los miocitos en normoxia y expuestos a lesión por isquemia simulada (80 y 100 veces más, n=5, p<0,05, n=3). Conclusión. Los resultados sugieren que el MITF-H podría estar involucrado en la hipertrofia, la respuesta al estrés por isquemia y la supervivencia de cardiomiocitos de cobayo.

Introduction: The microphthalmia -associated transcription factor ( MITF ) regulates the expression of specific genes and its cardiac expression and function is not known. Objectives: To identify the expression of MITF in hearts and isolated cardiomyocytes from Guinea pigs, to describe morphological changes associated with mRNA interference of MITF and to evaluate their relative changes in expression in isolated cardiomyocytes under ischemic preconditioning. Materials and methods: The cardiac specific isoform, MITF-H, and relative expression level analysis, was determined by semi-quantitative real time PCR, sequencing and Western blotting. Reduction of mRNA-MITF-H was induced by transduction of specific-MITF-shRNAi interference. The cardiac morphological changes, diameter and number of cardiac fibers were evaluated by direct observation and light microscopy. Results: A cDNA fragment of 281 bp was amplified from heart and isolated ventricular cardiac myocytes. Sequence analysis confirmed the identity of the isoform MITF-H, exon 1. The MITF silencing was associated with an increase in cardiac index (heart weight/body weight vs . 5.46 x 10 -3 vs 4.6 x 10 -3 ) and higher diameter of cardiac fibers (50.2±16 µ m vs 38,7±14,7 µ m p<0.05, n=150). In isolated cardiac myocytes under ischemic preconditioning we observed a higher relative expression compared with that measured in myocytes exposed to normoxia and simulated ischemia (eighty and one hundred times, p <0.05, n = 5). Conclusion. The results suggest that MITF-H isoform may be involved in Guinea pig cardiac hypertrophy, response to stress by ischemia and cardiomyocytes survival.

Paraoxonase1, its Q192R polymorphism and HDL-cholesterol in relation to intensive cardiac care unit stay in ischemic heart disease.

AIMS AND OBJECTIVES: The present study was evaluated the atheroprotective potential of paraoxonase1 (PON1) and its Q192R polymorphism, to determine whether this polymorphism, which is responsible for differential PON1 activity plays any role in the pathogenesis, severity and extent of coronary artery disease (CAD). MATERIALS AND METHODS: This hospital‑based cross‑sectional study investigated 60 diagnosed cases of CAD and 60 age and gender matched controls. All were assessed for serum PON1 activity, PON1 Q192R polymorphism and for classical cardiovascular risk factors. Individual serum phenotyping for PON1 Q192R polymorphism was determined by double substrate hydrolysis assay. Severity of CAD was assessed by the length of intensive cardiac care unit (ICCU) stay. RESULTS: Serum PON1 activity is significantly reduced in cases of CAD (92.6 ± 31.13 IU/L when compared with controls (105.26 ± 32.53 IU/L). Furthermore, serum arylesterase activity is reduced in CAD patients (90.31 ± 23.26 kU) when compared with the control subjects (101.61 ± 28.68 kU). Serum PON1 and arylesterase activities are significantly negatively correlated with the length of ICCU stay (r = −393 and r = −374 respectively). There is no significant difference in the occurrence of CAD and length of ICCU stay among the PON1 phenotypes (P = 0.92). Logistic regression analysis after adjustment of established risk factors revealed no significant association between CAD risk and PON1 Q192R polymorphism (odds ratios: 1.179 [95% confidence intervals: 0.507‑2.744], P = 0.702). SUMMARY AND CONCLUSIONS: The current study demonstrates that the activity of the PON1 enzyme may be more important factor than the PON1 Q192R polymorphism in the severity and extent of CAD.

Association between apolipoprotein E-polymorphism and ischemic heart disease patients with or without type 2 diabetes mellitus: a preliminary study in Kuwait

We investigated the association between apolipoprotein E polymorphism and ischemic heart disease with or without type 2 diabetes in Kuwait and examined the impact of apolipoprotein E polymorphism in diabetic patients. The present study was conducted from January 2005 to June 2006 in the Diabetic Clinic of AI-Amiri and Al-Sabah Hospitals in Kuwait City. Apolipoprotein E polymorphism was assessed in 250 subjects of which 83 were ischemic heart disease patients [41 diabetic and 42 non-diabetic] and 105 were diabetic patients without ischemic heart disease. Results were compared with 62 healthy controls. Apolipoprotein E polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism. Apolipoprotein E3 allele was the most commonly occurring form. The frequency of apolipoprotein E4 was higher in ischemic heart disease patients with type 2 diabetes [39%] and the non-diabetic [31%] group, but lower in the diabetic [20%] and control groups [16%]. Apolipoprotein E4 allele may be related to the development of ischemic heart disease in patients with or without type 2 diabetes in Kuwait. However, future studies with larger population sizes are needed to establish such relationship

Polimorfismo K121Q do gene ENPP1 e cardiopatia isquêmica em pacientes com diabete melito / Polimorfismo K121Q del gen ENPP1 y cardiopatía isquémica en pacientes con diabetes melitus / ENPP1 K121Q polymorphism and ischemic heart disease in diabetic patients

FUNDAMENTO: O gene ecto-nucleotídeo pirofosfatase/fosfodiesterase 1 (ENPP1) é um gene candidato à resistência insulínica. A resistência à insulina é um componente importante da síndrome metabólica e tem sido implicada no desenvolvimento de doença cardíaca isquêmica (DCI). OBJETIVO: Avaliar a associação entre o polimorfismo K121Q do gene ENPP1 e a presença da DCI em pacientes caucasianos com diabete melito (DM) tipo 2. MÉTODOS: Estudo transversal foi realizado em pacientes com DM tipo 2 (n=573; 50,6 por cento homens; idade 59,5±10,4 anos). DCI foi definida pela presença de angina ou infarto agudo do miocárdio pelo questionário cardiovascular da Organização Mundial da Saúde e/ou alterações compatíveis no ECG (código Minnesota) ou cintilografia miocárdica. O polimorfismo K121Q foi genotipado através da técnica de PCR e digestão enzimática. RESULTADOS: DCI esteve presente em 209 (36,5 por cento) pacientes. A frequência dos genótipos KK, KQ e QQ entre os pacientes com DCI foi 60,8 por cento, 34,4 por cento e 4,8 por cento, semelhante à distribuição dos genótipos entre os pacientes sem DCI (64,0 por cento, 32,7 por cento e 3,3 por cento, P = 0,574). Não se observou diferença nas características clínicas ou laboratoriais entre os três genótipos, nem em relação à presença de síndrome metabólica. CONCLUSÃO: Nenhuma associação foi encontrada entre o polimorfismo K121A do gene ENPP1 e a presença de DCI ou características fenotípicas de resistência insulínica.

BACKGROUND: The ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) gene is a candidate gene for insulin resistance. Insulin resistance is a major component of metabolic syndrome (MetS) and has been implicated in ischemic heart disease (IHD). OBJECTIVE: To evaluate the association between the K121Q polymorphism of the ENPP1 gene and IHD in white patients with type 2 diabetes mellitus (DM). METHODS: A cross-sectional study was performed in type 2 DM patients (n = 573, 50.6 percent males, age 59.5±10.4 years). IHD was defined by the presence of angina or myocardial infarction according to the Worth Health Organization cardiovascular questionnaire and/or compatible electrocardiographic (Minnesota Code), or perfusional abnormalities in myocardial scintigraphy. The K121Q polymorphism of ENPP1 gene was genotyped using PCR-based methods and restriction enzyme digestion. RESULTS: IHD was present in 209 (36.5 percent) patients. The distribution of KK, KQ and QQ genotypes among patients with IHD was 60.8 percent, 34.4 percent and 4.8 percent, not different from the genotype distribution in the group without IHD (64 percent, 32.7 percent and 3.3 percent, P=0.574). No difference was found in the clinical and laboratory characteristics between the three genotypes, neither regarding the prevalence of Metabolic Syndrome. CONCLUSION: No association was found between polymorphism K121A of ENPP1 gene and the presence of IHD.

FUNDAMENTO: El gen ecto-nucleótido pirofosfatasa/fosfodiesterasa 1 (ENPP1) es un gen candidato a la resistencia insulínica. La resistencia a la insulina es un componente importante del síndrome metabólico y ha sido involucrada en el desarrollo de enfermedad cardiaca isquémica (ECI). OBJETIVO: Evaluar la asociación entre el polimorfismo K121Q del gen ENPP1 y la presencia de ECI en pacientes caucásicos con diabetes melitus (DM) tipo 2. MÉTODOS: SE Realizó un estudio transversal en pacientes con DM tipo 2 (n=573; 50,6 por ciento hombres; edad 59,5±10,4 años). Se definió la ECI por la presencia de angina o infarto agudo de miocardio mediante el cuestionario cardiovascular de la Organización Mundial de la Salud y/o alteraciones compatibles en el ECG (código Minnesota) o centellograma miocárdico. El polimorfismo K121Q fue genotipificado mediante la técnica de PCR y digestión enzimática. RESULTADOS: La ECI estuvo presente en 209 (36,5 por ciento) pacientes. La frecuencia de los genotipos KK, KQ y QQ entrel os pacientes con ECI fue del 60,8 por ciento, 34,4 por ciento y 4,8 por ciento, semejante a la distribución de los genotipos entre los pacientes sin ECI (64,0 por ciento, 32,7 por ciento y 3,3 por ciento, P = 0,574). No se observó diferencia en las características clínicas o de laboratorio entre los tres genotipos, ni en relación con la presencia de síndrome metabólico. CONCLUSIÓN: No se encontró ninguna asociación entre el polimorfismo K121A del gen ENPP1 y la presencia de ECI o características fenotípicas de resistencia insulínica.

Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene

Myocardial ischemic preconditioning up-regulated protein 1 (Mipu1), a novel zinc finger protein, was originally cloned using bioinformatic analysis and 5' RACE technology of rat heart after a transient myocardial ischemia/reperfusion procedure in our laboratory. In order to investigate the functions of Mipu1, the recombinant prokaryotic expression vector pQE31-Mipu1 was constructed and transformed into Escherichia coli M15(pREP4), and Mipu1-6His fusion protein was expressed and purified. The identity of the purified protein was confirmed by mass spectrometry. The molecular mass of the Mipu1 protein was 70.03779 kDa. The fusion protein was intracutaneously injected to immunize New Zealand rabbits to produce a polyclonal antibody. The antibody titer was approximately 1:16,000. The antibody was tested by Western blotting for specificity and sensitivity. Using the antibody, it was found that Mipu1 was highly expressed in the heart and brain of rats and was localized in the nucleus of H9c2 myogenic cells. The present study lays the foundation for further study of the biological functions of Mipu1.

Correlación del polimorfismo (I/D) del gen de la ECA y la función ventricular en pacientes con miocardiopatía dilatada de origen isquémico e idiopático / ACE gene polymorphysm correlation (I/D) with the ventricular function in patients with ischemic and dilate idiopathic cardiomyopathy

Dilated cardiomyopathy is a myocardial disease, characterized by biventricular expansion. Renin-angiotensin-aldosterone system (RAAS) is closely related with the progress of this pathology. Has been shown that angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism influences as much in the plasmatic concentration as in activity of ACE. In addition, ACE I/D polymorphism has been associated with remodeling phenomena and an increased risk to develop several cardiovascular diseases. On virtue of the influence of ACE gene polymorphism on RAAS, we studied the correlation between ACE I/D polymorphism with morphologic and functional clinical alterations in ischemic or idiopathic dilated cardiomyopathy in one attempt to establish its utility as prognosis factor. Methods and results. We studied 30 patients of The National Institute of Cardiology. Ventricular function was evaluated by transthoracic echocardiography. ACE genotype was determined by polymerase chain reaction (PCR). Results for left ventricle shown: Tei Index was increased in patients with II genotype (0.84 vs. 0.48) when were compared to patients with DD genotype p < 0.01. Eccentricity Index was lesser in the group with II genotype (0.64), than in the group DD (0.86) p < 0.01. Ventricular mass was increased in DD patients when was compared with II group (174 g vs. 133 g) Isovolumetric contraction time was shorter in group DD than in II (45 mseg vs. 139 mseg) p < O.OB. These findings denote better preservation of left ventricular function in patients with DD genotype. In opposition, right ventricle shown an increased Tei Index in the group with DD genotype (1.01) when was compared with II genotype (0.55), p < 0.05. Pulmonary artery systolic pressure tended to be higher in DD genotype group without reach statistic significance. Conclusions. In our group of study, patients with DD genotype shown better left ventricular function in ischemia or idiopathic dilated cardiomyopathy. On the opposite right ventricular function were more deteriorated in patients with ACE DD genotype.

La miocardiopatía dilatada es una enfermedad primaria del miocardio, caracterizada por dilatación biventricular. El sistema renina-angiotensina-aldosterona (SRAA) está estrechamente relacionado con el progreso de esta patología. Se ha demostrado que el polimorfismo inserción/deleción (I/D) del gen de la enzima convertidora de angiotensina (ECA) influye en la concentración plasmática y la actividad de esta enzima, además este polimorfismo se ha asociado con fenómenos de remodelación e incremento en el riesgo de padecer diferentes enfermedades cardiovasculares. En virtud de la influencia de las variantes polimórficas del gen de la ECA sobre la respuesta del SRAA, en el presente trabajo se estudió la posible correlación del polimorfismo I/D del gen de la ECA con las alteraciones clínicas morfológicas y funcionales de la cardiomiopatía dilatada tanto de origen isquémico como de origen idiopático con el fin de establecer su posible utilidad como factor pronóstico. Métodos y resultados. El estudio incluyó a 30 pacientes seleccionados de la consulta externa del Instituto Nacional de Cardiología <>, la función ventricular se valoró mediante ecocardiografía transtorácica. El genotipo de la ECA se determinó por reacción en cadena de la polimerasa (PCR). Resultados para el ventrículo izquierdo: El índice de Tei se observó visiblemente incrementado en los pacientes con genotipo II 0.84 vs. 0.48 de los pacientes con genotipo DD p < 0.01. El índice de excentricidad fue menor en los casos con genotipo II: 0.64, comparado con aquellos con genotipo DD: 0.86 p < 0.01. La masa ventricular tendió a ser mayor en el grupo DD en relación con el II (174 g vs. 133 g). El tiempo de contracción isovolumétrica fue menor en el grupo DD en comparación al II (45 mseg vs. 139 mseg) p < 0.05, estos hallazgos denotan una mejor preservación de la función ventricular izquierda en los pacientes con genotipo DD. Por otra parte, el ventrículo derecho mostró un comportamiento distinto al observado para el ventrículo izquierdo, pues el índice de Tei fue mayor para los pacientes con el genotipo DD (1.01) comparado con el grupo del genotipo II (0.55), p < 0.05. La presión sistólica de la arteria pulmonar tendió a ser mayor en los pacientes con genotipo DD sin alcanzar una significancia estadística. Conclusión. El genotipo DD se asocia con una mejor función ventricular izquierda en los pacientes con miocardiopatía dilatada de origen tanto isquémico como idiopático; por el contrario, la función ventricular derecha de los pacientes con genotipo DD muestra una mayor alteración en el índice de Tei en esta patología.

El papel del factor de transcripción NF-κB en la célula cardíaca / Role of the transcription factor NF-κB in the cardiac cell

Para la biología de hoy las vías de señalización intracelular que controlan los procesos entre la vida y la muerte celular son de gran interés. Al respecto, el NF-κB destaca como un factor de transcripción decisivo de respuesta rápida que participa en la activación de las vías de señalización de la muerte celular programada. Lo relevante es que sus efectos tienen consecuencias en el desarrollo normal y/o la homeostasis en muchas células o tejidos, que incluyen entre otros al sistema inmune, los folículos capilares, apéndices epidermales, el riñon y el sistema nervioso. En esta revisión analizamos el papel central que juega el factor de transcripción NF-κB en el funcionamiento normal de la célula cardíaca y sus implicaciones en algunas de las patologías cardíacas más frecuentes como: el daño por isquemia-reperfusión, la isquemia precondicionada, la hipertrofia, la aterosclerosis, y el paro cardíaco. El NF-κB comúnmente funciona como un agente citoprotector, aunque hay algunos casos en los cuales resulta ser pro-apoptótico dependiendo del estímulo y del contexto celular. Se han logrado avances significativos a nivel molecular, que han permitido entender su modo de acción y el papel interactivo que juega con otros factores claves. Estos estudios han identificado muchos genes anti-apoptóticos y pro-apoptóticos regulados por la actividad del NF-κB abriendo novedosas aproximaciones que se pueden hacer sobre sus efectos en el desarrollo de patologías cardíacas.

The signaling pathways that control the life-death switch of a cell are a prime interest in Modern Biology. To this respect, NF-κB has emerged as a decisive transcription factor in the cell's response to apoptotic challenge and its effects on apoptosis have far-reaching consequences for normal development and/or homeostasis in many cells and tissues, including the immune system, hair follicles, and epidermal appendages, the liver, and nervous system. In this review we analyze the pivotal role of the transcription factor NF-κB in the normal functioning of the cardiac cell and its implication on some of the most frequent cardiac pathologies, such as ischemia-reperfusion injury, ischemic precondition, hypertrophy, atherosclerosis and cardiac arrest. While NF-κB is commonly found to be cytoprotective, there are a number of instances where it is proapoptotic depending on the inducing stimulus and the cell context. Significant progress has been made in understanding its mode of action and its interplay with other key factors. These studies identified many anti- and pro-apoptotic NF-κB regulated genes that mediate its activity, these important new insights fuel hope that novel approaches will be developed to control the effects of NF-κB in cardiac pathologies.

Actuales factores de riesgo aterogénico en la génesis de la cardiopatía isquémica: problemática epidemiológica mundial / Current atherogenic risk factors in the genesis of ischemic heart disease

Se presentaron las principales tasas de morbilidad-mortalidad por cardiopatía isquémica (CI) en algunos países del mundo incluida Cuba, lo que constituye una gran problemática epidemiológica universal. Se expusieron y clasificaron los actuales factores de riesgo aterogénicos y coronarios, en la génesis de la cardiopatía isquémica

Representational difference analysis of cDNA identifies novel genes expressed following preconditioning of the heart

Preconditioning of the myocardium rapidly induces a number of transcription factors, which are likely to be responsible for a cascade of transcriptional changes underlying the development of delayed adaptation. Identifying these changes provides insight into the molecular pathways elicited by sub-lethal ischaemia and the mechanism leading to delayed adaptation. Genes up-regulated in rabbit myocardium in vivo by ischaemic preconditioning following reperfusion for 2 h, 4 h and 6 h posttreatment were identified by representational difference analysis of cDNA (cDNA. RDA). The area of the left ventricle rendered ischaemic by preconditioning or the equivalent area of sham-treated animals was isolated and cDNA.RDA performed. Three novel genes and six genes with known function where identified, including the TGFbeta receptor interacting protein 1, the alpha isoform of the A subunit of PP2 and the cap binding protein NCBP1. To determine whether expression of these genes correlated with preconditioning per se, expression was measured in myocardium after both ischaemic as well as heat shock induced preconditioning following 2 h, 4 h, and 6 h reperfusion. These genes were induced in rabbit myocardium in vivo by both ischaemia and heat shock, consistent with a fundamental role in the development of delayed adaptation. The well described role of PP2 in modulating the mitogen-activated protein kinase pathway and promoting cell survival is consistent with our previous work, which identified the reperfusion injury salvage kinase pathway in mediating the protective effects of ischaemic preconditioning. Expression of Trip1 and Ncbp1 also implicates TGFbeta signalling pathways and RNA processing and transport in delayed adaptation to stress in the myocardium.

Intramyocardial gene transfer of vascular endothelial growth factor 121 improves myocardial perfusion and function in the ischemic porcine heart

Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen that is angiogenic in vitro and in vivo. Several studies report on gene transfer of VEGF121 to promote angiogenesis in the ischemic myocardium of animals and patients. We hypothesized that intramyocardial administration of naked plasmid DNA encoding VEGF121 could improve myocardial perfusion and function in a porcine model of myocardial ischemia. Yorkshire swine underwent thoracotomy and placement of an ameroid constrictor on the circumflex coronary artery. Four weeks later, pVEGF121 plasmid was administered into the ischemic myocardium. Four weeks after gene transfer, SPECT imaging demonstrated significant reduction in the ischemic area in pVEGF121-treated animals compared with controls. In the pVEGF121 group, most of the animals evolved from light ischemia to a normal perfusion. In contrast, control animals exhibited similar or impaired ischemic conditions. Our results indicate that intramyocardial gene transfer of VEGF121 as naked plasmid DNA results in significant improvement in myocardial perfusion and function.

The research progress of the association of mitochondrial DNA mutation with cardiomyopathy / 法医学杂志

There are some human diseases associated with mitochondrial DNA genome defect. Now many studies think that: oxygen radical resulting from oxidative phosphorylation(OXPHOS) disorder caused by myocardium ischemia and the increased OXPHOS induction damage mitochondrial DNA. Chronic damage accumulations lead to mitochondrial DNA deletion or point mutation in the end which show mitochondrial DNA 5.0 kb or 7.4 kb deletion and point mutation at position C15452A in the cytochrome b gene; the conservative sequence mutation of tRNA gene such as A4300G, C4320T point mutation in the tRNA Ilegene, A3243G point mutation in the tRNA leu gene etc result in defective contractile proteins whose persistent and inefficient contraction may increase the myocardium's metabolic demands for ATP and leads to cardiac hypertrophy. In this article, we review the study on the association of mitochondrial DNA mutation with ischemic cardiomyopathy and hypertrophic cardiomyopathy.