Crataegus Extract WS®1442 Stimulates Cardiomyogenesis and Angiogenesis From Stem Cells: A Possible New Pharmacology for Hawthorn?

Extracts from the leaves and flowers of Crataegus spp. (i.e., hawthorn species) have been traditionally used with documented preclinical and clinical activities in cardiovascular medicine. Based on reported positive effects on heart muscle after ischemic injury and the overall cardioprotective profile, the present study addressed potential contributions of Crataegus extracts to cardiopoietic differentiation from stem cells. The quantified Crataegus extract WS®1442 stimulated cardiomyogenesis from murine and human embryonic stem cells (ESCs). Mechanistically, this effect was found to be induced by promoting differentiation of cardiovascular progenitor cell populations but not by proliferation. Bioassay-guided fractionation, phytochemical and analytical profiling suggested high-molecular weight ingredients as the active principle with at least part of the activity due to oligomeric procyanidines (OPCs) with a degree of polymerization between 3 and 6 (DP3–6). Transcriptome profiling in mESCs suggested two main, plausible mechanisms: These were early, stress-associated cellular events along with the modulation of distinct developmental pathways, including the upregulation of brain-derived neurotrophic factor (BDNF) and retinoic acid as well as the inhibition of transforming growth factor β/bone morphogenetic protein (TGFβ/BMP) and fibroblast growth factor (FGF) signaling. In addition, WS®1442 stimulated angiogenesis ex vivo in Sca-1+ progenitor cells from adult mice hearts. These in vitro data provide evidence for a differentiation promoting activity of WS®1442 on distinct cardiovascular stem/progenitor cells that could be valuable for therapeutic heart regeneration after myocardial infarction. However, the in vivo relevance of this new pharmacological activity of Crataegus spp. remains to be investigated and active ingredients from bioactive fractions will have to be further characterized

Sex-Specific associations of brain-derived neurotrophic factor and cardiorespiratory fitness in the general population

The brain-derived neurotrophic factor (BDNF) was initially considered to be neuron-specific. Meanwhile, this neurotrophin is peripherally also secreted by skeletal muscle cells and increases due to exercise. Whether BDNF is related to cardiorespiratory fitness (CRF) is currently unclear. We analyzed the association of serum BDNF levels with CRF in the general population (Study of Health in Pomerania (SHIP-TREND) from Northeast Germany; n = 1607, 51% female; median age 48 years). Sex-stratified linear regression models adjusted for age, height, smoking, body fat, lean mass, physical activity, and depression analyzed the association between BDNF and maximal oxygen consumption (VO2peak), maximal oxygen consumption normalized for body weight (VO2peak/kg), and oxygen consumption at the anaerobic threshold (VO2@AT). In women, 1mL/min higher VO2peak, VO2peak/kg, and VO2@AT were associated with a 2.43 pg/mL (95% confidence interval [CI]: 1.16 to 3.69 pg/mL; p = 0.0002), 150.66 pg/mL (95% CI: 63.42 to 237.90 pg/mL; p = 0.0007), and 2.68 pg/mL (95% CI: 0.5 to 4.8 pg/mL; p = 0.01) higher BDNF serum concentration, respectively. No significant associations were found in men. Further research is needed to understand the sex-specific association between CRF and BDNF. © 2019 by the authors. Licensee MDPI, Basel, Switzerland

Sex-Specific Associations of Brain-Derived Neurotrophic Factor and Cardiorespiratory Fitness in the General Population

The brain-derived neurotrophic factor (BDNF) was initially considered to be neuron-specific. Meanwhile, this neurotrophin is peripherally also secreted by skeletal muscle cells and increases due to exercise. Whether BDNF is related to cardiorespiratory fitness (CRF) is currently unclear. We analyzed the association of serum BDNF levels with CRF in the general population (Study of Health in Pomerania (SHIP-TREND) from Northeast Germany; n = 1607, 51% female; median age 48 years). Sex-stratified linear regression models adjusted for age, height, smoking, body fat, lean mass, physical activity, and depression analyzed the association between BDNF and maximal oxygen consumption (VO2peak), maximal oxygen consumption normalized for body weight (VO2peak/kg), and oxygen consumption at the anaerobic threshold (VO2@AT). In women, 1mL/min higher VO2peak, VO2peak/kg, and VO2@AT were associated with a 2.43 pg/mL (95% confidence interval [CI]: 1.16 to 3.69 pg/mL; p = 0.0002), 150.66 pg/mL (95% CI: 63.42 to 237.90 pg/mL; p = 0.0007), and 2.68 pg/mL (95% CI: 0.5 to 4.8 pg/mL; p = 0.01) higher BDNF serum concentration, respectively. No significant associations were found in men. Further research is needed to understand the sex-specific association between CRF and BDNF. © 2019 by the authors. Licensee MDPI, Basel, Switzerland

Einfluss von Cerivastetin auf die Expression von eNOS und LOX-1 in humanen venösen Endothelzellen

Die vorliegende Arbeit behandelt den Einfluss von Cerivastatin auf die TNF-alpha-modulierte Expression von eNOS (endotheliale NO-Synthase) und LOX-1, dem Rezeptor für oxidiertes LDL, in humanen venösen Endothelzellen. Sie beinhaltet die Untersuchung der mRNA- und Proteinebene, dafür erfolgt die Generierung eines Antikörpers gegen LOX-1. Im Alamar-blue-Test zeigt sich keine physiologisch relevante Abnahme der Viabilität der behandelten Zellen. Die Inkubation mit TNF-alpha führt zu inversen Effekten auf die Expression von eNOS und LOX-1. Es zeigt sich eine Minderung der eNOS-mRNA- und Proteinexpression, die durch Cerivastatin reversibel ist. Die Statinwirkung ist auf mRNA-Ebene vollständig, auf Proteinebene teilweise durch Mevalonat antagonisierbar. Für LOX-1 zeigt sich eine konzentrations- und zeitabhängige Steigerung der LOX-1 -mRNA-und Proteinexpression nach Stimulierung mit TNF-alpha. Nach 8-stündiger Inkubation mit Cerivastatin kommt es zu einer Minderung der LOX-1-mRNA- und Proteinexpression im Vergleich zu zytokin-stimulierten Zellen. Nach 12 Stunden führen niedrigere Konzentrationen Cerivastatin zu einer geringen Abnahme der der mRNA-Expression, während höhere Konzentrationen die Expression signifikant steigern. Unter allen Cerivastatinkonzentrationen kommt es im Trend zu einer Zunahme der LOX-1 Proteinexpression in TNF-alpha-stimulierten Zellen. Die Effekte des Statins auf LOX-1 sind nur zum Teil durch Mevalonat antagonisierbar.keine Angabe

Simultaneous assessment of spontaneous cage activity and voluntary wheel running in group-housed mice

Small animal models are frequently used to improve our understanding of the molecular and biological signaling pathways underlying the beneficial effects of physical activity and exercise. Unfortunately, when running wheels are employed, mice and rats are often kept single-housed to determine the individual running distance of each animal. However, social isolation can be stressful for rodents, and may alter an individual’s propensity for or response to exercise. For example, increased stress from single housing may significantly affect the results when investigating systemic metabolic responses to exercise. We have combined two already available and well-established systems, a radiotelemetry system and a running wheel, to determine spontaneous cage activity (SCA) as well as voluntary exercise (VE) levels of the individual animal in group-housed rodents. Further, we developed a simple software tool which allows monitoring and analyzing the data. Specifically, the radiotelemetry-system utilizes radio-frequency identification via a small, implanted chip to determine the location of each animal. Since, in addition to the animals’ position, also the location of the running wheel in the cage is known, the conclusion of which animal is exercising can be drawn. The developed software enables a fast and reliable assignment of the VE data to the individual animal and a simple analysis of the data collected. Hence, our combined method may be used to investigate the beneficial effects of physical activity, as well as the impact of therapeutic interventions on animal behavior in group-housed rodents

A Transgenic Mouse Model of Eccentric Left Ventricular Hypertrophy With Preserved Ejection Fraction Exhibits Alterations in the Autophagy-Lysosomal Pathway

The ubiquitin-proteasome system (UPS) and the autophagy-lysosomal pathway (ALP) are the main proteolytic systems involved in cellular homeostasis. Since cardiomyocytes, as terminally differentiated cells, lack the ability to share damaged proteins with their daughter cells, they are especially reliant on these protein degradation systems for their proper function. Alterations of the UPS and ALP have been reported in a wide range of cardiac diseases, including cardiomyopathies. In this study, we determined whether the UPS and ALP are altered in a mouse model of eccentric left ventricular (LV) hypertrophy expressing both cyclin T1 and Gαq under the control of the cardiac-specific α-myosin heavy chain promoter (double transgenic; DTG). Compared to wild-type (WT) littermates, DTG mice showed higher end-diastolic (ED) LV wall thicknesses and diameter with preserved ejection fraction (EF). The cardiomyopathic phenotype was further confirmed by an upregulation of the fetal gene program and genes associated with fibrosis as well as a downregulation of genes involved in Ca2+ handling. Likewise, higher NT-proBNP levels were detected in DTG mice. Investigation of the UPS showed elevated steady-state levels of (poly)ubiquitinated proteins without alterations of all proteasomal activities in DTG mice. Evaluation of ALP key marker revealed a mixed pattern with higher protein levels of microtubule-associated protein 1 light chain 3 beta (LC3)-I and lysosomal-associated membrane protein-2, lower protein levels of beclin-1 and FYVE and coiled-coil domain-containing protein 1 (FYCO1) and unchanged protein levels of p62/SQSTM1 in DTG mice when compared to WT. At transcriptional level, a > 1.2-fold expression was observed for Erbb2, Hdac6, Lamp2, Nrg1, and Sqstm1, while a < 0.8-fold expression was revealed for Fyco1 in DTG mice. The results related to the ALP suggested overall a repression of the ALP during the initiation process, but an induction of the ALP at the level of autophagosome-lysosome fusion and the delivery of ubiquitinated cargo to the ALP for degradation

Preliminary results of the cross-sectional associations of sedentary behavior and physical activity with serum brain-derived neurotrophic factor in adults with coronary heart disease

This is the first study to analyze the association of accelerometer-measured patterns of habitual physical activity (PA) and sedentary behavior (SB) with serum BDNF in individuals with coronary heart disease. A total of 30 individuals (M = 69.5 years; 80% men) participated in this pre-post study that aimed to test a multi-behavioral intervention. All participants underwent standardized measurement of anthropometric variables, blood collection, self-administered survey, and accelerometer-based measurement of PA and SB over seven days. Serum BDNF concentrations were measured using enzyme-linked immunosorbent assay kit. We applied separate multiple linear regression analysis to estimate the associations of baseline SB pattern measures, light and moderate-to-vigorous PA with serum BDNF (n = 29). Participants spent 508.7 ± 76.5 min/d in SB, 258.5 ± 71.2 min/d in light PA, and 21.2 ± 15.2 min/d in moderate-to-vigorous PA. Per day, individuals had 15.5 ± 3.2 numbers of 10-to-30 min bouts of SB (average length: 22.2 ± 2.1 min) and 3.4 ± 1.2 numbers of > 30 min bouts of SB (average length: 43.8 ± 2.4 min). Regression analysis revealed no significant associations between any of the accelerometer-based measures and serum BDNF. The findings of this study did not reveal an association of accelerometer-measured PA and SB pattern variables with serum BDNF in individuals with coronary heart disease. In addition, our data revealed a considerable variation of PA and SB which should be considered in future studies

Brain Derived Neurotrophic Factor Contributes to the Cardiogenic Potential of Adult Resident Progenitor Cells in Failing Murine Heart

<div><p>Aims</p><p>Resident cardiac progenitor cells show homing properties when injected into the injured but not to the healthy myocardium. The molecular background behind this difference in behavior needs to be studied to elucidate how adult progenitor cells can restore cardiac function of the damaged myocardium. Since the brain derived neurotrophic factor (BDNF) moderates cardioprotection in injured hearts, we focused on delineating its regulatory role in the damaged myocardium.</p><p>Methods and Results</p><p>Comparative gene expression profiling of freshly isolated undifferentiated Sca-1 progenitor cells derived either from heart failure transgenic αMHC-CyclinT1/Gαq overexpressing mice or wildtype littermates revealed transcriptional variations. Bdnf expression was up regulated 5-fold during heart failure which was verified by qRT-PCR and confirmed at protein level. The migratory capacity of Sca-1 cells from transgenic hearts was improved by 15% in the presence of 25ng/ml BDNF. Furthermore, BDNF-mediated effects on Sca-1 cells were studied via pulsed Stable Isotope Labeling of Amino acids in Cell Culture (pSILAC) proteomics approach. After BDNF treatment significant differences between newly synthesized proteins in Sca-1 cells from control and transgenic hearts were observed for CDK1, SRRT, HDGF, and MAP2K3 which are known to regulate cell cycle, survival and differentiation. Moreover BDNF repressed the proliferation of Sca-1 cells from transgenic hearts.</p><p>Conclusion</p><p>Comparative profiling of resident Sca-1 cells revealed elevated BDNF levels in the failing heart. Exogenous BDNF (i) stimulated migration, which might improve the homing ability of Sca-1 cells derived from the failing heart and (ii) repressed the cell cycle progression suggesting its potency to ameliorate heart failure.</p></div

BDNF enhances migration of progenitor cells.

<p>Migration of Sca-1 cells (4 x 10⁴) was evaluated using a modified Boyden chamber assay. Freshly isolated cells were exposed to varying concentrations of BDNF (10, 25, 50ng/ml) and allowed to migrate for 2 hours (n = 3; mean ± SD; ANOVA; ***<i>p</i> < 0.0001, **<i>p</i> < 0.001, *<i>p</i> < 0.05 vs Control; §§<i>p</i> < 0.001, §<i>p</i> < 0.05 vs BDNF).</p

Expression of BDNF and receptor TrkB on progenitor cells.

<p>(A) Immunoblot data depicted higher expression of BDNF in freshly isolated Cyc cells in comparison to Wt. Bar graph indicates relative protein abundance of BDNF normalized to ß-actin control quantified by densitometry (n = 3; mean ± SD; *<i>p</i><0.05; t-test). (B) mRNA intensity signals of TrkB receptor determined by microarray analysis (n = 2; mean ± SD). (C-D) Immunofluorescence micrographs display the expression of TrkB receptor on Sca-1 cells. Nuclei were stained with DAPI. (Scale bar: 50μm, image magnification: 40x). (E) Protein abundance of TrkB receptor on Sca-1 cells quantified against ß-actin control by densitometry (n = 3, mean ± SD). Representative immunoblot of both full length and truncated forms of TrkB receptor on Wt and Cyc cells is shown below.</p