scholarly journals Rapid Induction of Aldosterone Synthesis in Cultured Neonatal Rat Cardiomyocytes under High Glucose Conditions

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Masami Fujisaki ◽  
Tomohisa Nagoshi ◽  
Tetsuo Nishikawa ◽  
Taro Date ◽  
Michihiro Yoshimura
Keyword(s):  
High Glucose ◽  
Culture System ◽  
Cardiac Tissue ◽  
Current System ◽  
Experimental Models ◽  
Neonatal Rat ◽  
Angiotensin Ii Receptor Blocker ◽  
Angiotensin Ii Receptor ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes

In addition to classical adrenal cortical biosynthetic pathway, there is increasing evidence that aldosterone is produced in extra-adrenal tissues. Although we previously reported aldosterone production in the heart, the concept of cardiac aldosterone synthesis remains controversial. This is partly due to lack of established experimental models representing aldosterone synthase (CYP11B2) expression in robustly reproducible fashion. We herein investigated suitable conditions in neonatal rat cardiomyocytes (NRCMs) culture system producing CYP11B2 with considerable efficacy. NRCMs were cultured with various glucose doses for 2–24 hours. CYP11B2 mRNA expression and aldosterone concentrations secreted from NRCMs were determined using real-time PCR and enzyme immunoassay, respectively. We found that suitable conditions for CYP11B2 induction included four-hour incubation with high glucose conditions. Under these particular conditions, CYP11B2 expression, in accordance with aldosterone secretion, was significantly increased compared to those observed in the cells cultured under standard-glucose condition. Angiotensin II receptor blocker partially inhibited this CYP11B2 induction, suggesting that there is local renin-angiotensin-aldosterone system activation under high glucose conditions. The suitable conditions for CYP11B2 induction in NRCMs culture system are now clarified: high-glucose conditions with relatively brief period of culture promote CYP11B2 expression in cardiomyocytes. The current system will help to accelerate further progress in research on cardiac tissue aldosterone synthesis.

Abstract 063: Rapid Induction of Aldosterone Synthesis in Cultured Neonatal Rat Cardiomyocytes under High Glucose Conditions

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Tomohisa Nagoshi ◽  
Masami Fujisaki ◽  
Tetsuo Nishikawa ◽  
Taro Date ◽  
Michihiro Yoshimura
Keyword(s):  
High Glucose ◽  
Culture System ◽  
Cardiac Tissue ◽  
Current System ◽  
Experimental Models ◽  
Neonatal Rat ◽  
Aldosterone Secretion ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes ◽  
Aldosterone Production

Background: In addition to the classical adrenal cortical biosynthetic pathway, there is increasing evidence that aldosterone is produced in extra-adrenal tissues. Although we have previously reported that aldosterone is produced in the heart, the concept of cardiac aldosterone synthesis remains controversial. This is partly due to the lack of established experimental models representing the aldosterone synthase gene (CYP11B2) expression in a robustly reproducible fashion. Therefore, in this study, we investigated suitable conditions in a neonatal rat cardiomyocytes culture system producing CYP11B2 with considerable efficacy. Methods and Results: Neonatal rat cardiomyocytes were cultured in serum-free medium containing various doses of glucose for two to 24 hours. The CYP11B2 mRNA expression in the cardiomyocytes was determined using real-time PCR, and the aldosterone concentrations secreted from the cardiomyocytes into the culture medium were measured using an enzyme immunoassay. We found that suitable conditions for inducing the cardiac aldosterone gene expression included four hours of incubation with high glucose conditions (>25 mM). Under these particular conditions, the CYP11B2 expression (2.18±0.37-fold, p<0.01), in accordance with aldosterone secretion, was significantly increased compared to that observed in the cells cultured under standard glucose condition (5.5 mM). Conclusions: The suitable conditions for detecting the CYP11B2 expression and aldosterone secretion in the neonatal rat cardiomyocytes culture system are now clarified: high glucose conditions with a relatively brief period of culture promote the CYP11B2 expression as well as aldosterone production/secretion in cardiomyocytes. The current system will help to accelerate further progress in research on cardiac tissue aldosterone synthesis.

Radicicol activates heat shock protein expression and cardioprotection in neonatal rat cardiomyocytes

2004 ◽  
Vol 287 (3) ◽  
pp. H1081-H1088 ◽  
Author(s):  
Tina M. Griffin ◽  
Tina V. Valdez ◽  
Ruben Mestril
Keyword(s):  
Heat Shock ◽  
Transcriptional Activation ◽  
Cardiac Tissue ◽  
Defense Mechanism ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Neonatal Rat ◽  
Heat Shock Factor 1 ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes

Heat shock proteins (HSPs) constitute an endogenous cellular defense mechanism against environmental stresses. In the past few years, studies have shown that overexpression of HSPs can protect cardiac myocytes against ischemia-reperfusion injury. In an attempt to increase the HSPs in cardiac tissue, we used the compound radicicol that activates HSP expression by binding to the HSP 90 kDa (HSP90). HSP90 is the main component of the cytosolic molecular chaperone complex, which has been implicated in the regulation of the heat shock factor 1 (HSF1). HSF1 is responsible for the transcriptional activation of the heat shock genes. In the present study, we show that radicicol induces HSP expression in neonatal rat cardiomyocytes, and this increase in HSPs confers cardioprotection to these cardiomyocytes. We also show that radicicol induction of the HSP and cardioprotection is dependent on the inhibition of HSP90 in cardiomyocytes. These results indicate that modulation of the active HSP90 protein level plays an important role in cardioprotection. Therefore, compounds, such as radicicol and its possible derivatives that inhibit the function of HSP90 in the cell may represent potentially useful cardioprotective agents.

Profiling of skeletal muscle Ankrd2 protein in human cardiac tissue and neonatal rat cardiomyocytes

2015 ◽  
Vol 143 (6) ◽  
pp. 583-597 ◽  
Author(s):  
Jovana Jasnic-Savovic ◽  
Aleksandra Nestorovic ◽  
Slobodan Savic ◽  
Sinisa Karasek ◽  
Nicola Vitulo ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Cardiac Tissue ◽  
Neonatal Rat ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes ◽  
Human Cardiac Tissue

scholarly journals Curcumin Protects Neonatal Rat Cardiomyocytes against High Glucose-Induced Apoptosis via PI3K/Akt Signalling Pathway

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Wei Yu ◽  
Wenliang Zha ◽  
Zhiqiang Ke ◽  
Qing Min ◽  
Cairong Li ◽  
...  
Keyword(s):  
Nadph Oxidase ◽  
High Glucose ◽  
Primary Cultures ◽  
Signalling Pathway ◽  
Neonatal Rat ◽  
Curcumin Treatment ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes ◽  
Enhanced Expression ◽  
Induced Apoptosis

The function of curcumin on NADPH oxidase-related ROS production and cardiac apoptosis, together with the modulation of protein signalling pathways, was investigated in cardiomyocytes. Primary cultures of neonatal rat cardiomyocytes were exposed to 30 mmol/L high glucose with or without curcumin. Cell viability, apoptosis, superoxide formation, the expression of NADPH oxidase subunits, and potential regulatory molecules, Akt and GSK-3β, were assessed in cardiomyocytes. Cardiomyocytes exposure to high glucose led to an increase in both cell apoptosis and intracellular ROS levels, which were strongly prevented by curcumin treatment (10 μM). In addition, treatment with curcumin remarkably suppressed the increased activity of Rac1, as well as the enhanced expression ofgp91phoxandp47phoxinduced by high glucose. Lipid peroxidation and SOD were reversed in the presence of curcumin. Furthermore, curcumin treatment markedly inhibited the reduced Bcl-2/Bax ratio elicited by high glucose exposure. Moreover, curcumin significantly increased Akt and GSK-3βphosphorylation in cardiomyocytes treated with high glucose. In addition, LY294002 blocked the effects of curcumin on cardiomyocytes exposure to high glucose. In conclusion, these results demonstrated that curcumin attenuated high glucose-induced cardiomyocyte apoptosis by inhibiting NADPH-mediated oxidative stress and this protective effect is most likely mediated by PI3K/Akt-related signalling pathway.

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