The effect of fructose rich diet on the expression of renin-angiostensin system components and inflamatory molecules in the rat heart: sex specific differences

Ishrana bogata fruktozom predstavlja bitan faktor u razvoju metaboličkog sindroma koji je povezan sa povećanim rizikom za nastanak kardiovaskularnih bolesti. Smatra se da hronična inflamacija ima bitnu ulogu u njegovoj patogenezi. Ženke su zaštićene od ishranom izazvanih metaboličkih poremećaja i hipertenzije u reproduktivnom periodu. Estrogen, koji utiče na normalno funkcionisanje kardiovaskularnog sistema, bi mogao doprinositi uočenim polno specifičnim razlikama u nastanku simptoma metaboličkog sindroma. Proteinske komponente renin-angiotenzin sistema (RAS) imaju bitnu ulogu u nastanku inflamacije, hipertenzije i insulinske rezistencije i na njihovu ekspresiju utiče estradiol. Ova doktorska disertacija je za cilj imala da ispita da li pol i estradiol doprinose promenama u ekspresiji komponenti RAS-a [angiotenzin konvertujućeg enzima (ACE), angiotenzin konvertujućeg enzima 2 (ACE2), angiotenzinskog receptora tipa 1 (AT1R), angiotenzinskog receptora tipa 2 (AT2R) i kolektrina] i medijatora inflamacije i remodelovanja tkiva srca [nuklearnog faktora κB (NFκB), matriks metaloproteinaze-9 (MMP-9) i liganda 16 iz familije hemokina CXC (CXCL16)] kod animalnog modela metaboličkog sindroma. Mužjaci i ženke pacova koji su pili 10% rastvor fruktoze umesto vode u trajanju od 9 nedelja predstavljali su animalni model metaboličkog sindroma korišćen u ovoj studiji. Radi ispitivanja efekata estradiola jedan deo ženki pacova je ovarijektomisan i podeljen u tri grupe pri čemi je jedna grupa pored standardne laboratorijske hrane pila vodu, druga grupa je pila 10% rastvor fruktoze, a treća grupa je pored rasvora fruktoze primala i supstitucionu terapiju estradiolom. Ishrana bogata fruktozom je uzrokovala povećanje krvnog pritiska samo kod mužjaka pacova. Ovaj tip ishrane nije doveo do promena u masi srca niti u odnosu masa srca/masa tela ni kod jednog pola, kao ni kod ovarijektomisanih životinja što ukazuje da se hipertrofija srca nije razvila...Fructose rich diet (FRD) represents an important factor in the development of metabolic syndrome. Metabolic syndrome is associated with increased risk for cardiovascular diseases occurrence and chronic inflammation has a major role in its pathogenesis. Females are protected from diet-induced metabolic disturbances and hypertension in their reproductive period. Estrogen, which influences the cardiovascular system, could contribute to the observed gender-specific differences in the onset of metabolic syndrome. Components of the renin-angiotensin system (RAS), the synthesis of which is mediated by estrogen, have an important role in the inflammatory processes, hypertension and insulin resistance. Almost all of the components of metabolic syndrome increase the activity of RAS, which finally results in increased oxidative stress and inflammation. The aim of this doctoral dissertation was to investigate sex specific changes and role of estradiol in the expression of RAS components [angiotensin converting enzyme (ACE), angiotensin converting enzyme 2 (ACE2), angiotensin receptor type 1 (AT1R), angiotensin receptor type 2 (AT2R) and collectrin], as well as mediators of inflammation and remodeling of heart tissue [nuclear faktor κB (NFκB), matrix metalloproteinse-9 (MMP-9) and CXCL16 chemokine] in the animal model of metabolic syndrome. Male and female rats, which consumed 10% fructose solution instead of water for 9 weeks, represent an animal model of metabolic syndrome that was used in this study. In order to examine the effects of estradiol in the context of FRD, female rats were ovariectomized and divided in three groups: fed normal diet, fed FRD, and fed FRD and subjected to estradiol replacement therapy. FRD increased blood pressure only in male rats. This diet regime didn't cause heart hypertrophy neither in intact males and females, nor in ovariectomized females..

Ekspresija komponenti renin-angiotenzin sistema u humanom karotidnom plaku

Background/Aim. The renin-angiotensin system (RAS) is linked to the development of atherosclerosis (As), including its initiation and progression. Besides the well-known angio-tensin-converting enzyme (ACE), two newer RAS family members are related to vascular remodeling - ACE2 as a homolog of ACE and collectrin [transmembrane protein 27 (TMEM27)] as a homolog of ACE2. Up to now, a limited number of studies have examined the expression of these RAS components in advanced carotid plaque (CP) tissue based on the sex of the patients and plaque phenotypes (PPs). There are two ultrasonographically defined PPs - the hypoechogenic plaque (HoP) and the hyperechogenic plaque (HerP) phenotype. The aim of the study was to investigate whether there was a correlation between the expression of RAS components in the CP and the sex and PPs of patients. Methods. We examined 74 patients with advanced CP who underwent carotid endarterectomy. The intraplaque expression of RAS components was determined with the real-time polymerase chain reaction, using the TaqMan® gene expression assays and Western blot. A two-way ANOVA followed by a post-hoc Tukey test was performed for the statistical analysis of results. Results. No interaction was recorded between the sex of the patients and PPs in influencing the relative expression of ACE and TMEM27 messenger RNA (mRNA) (p > 0.05). In 56.06% of plaque samples, no expression of ACE2 mRNA was detected. Among the plaques where ACE2 mRNA expression was detected, its expression level was higher in females with the HoP phenotype compared to females with the HerP phenotype (p < 0.001). In patients with the HoP phenotype, females had higher expression of ACE2 mRNA than males (p < 0.05). In the male study group, ACE protein levels were significantly lower in the HoP phenotype compared to the HerP phenotype (p < 0.001). Fe-males with the HoP phenotype had significantly higher ACE protein levels than males with the HoP phenotype (p < 0.0001). Conclusion. Our results revealed alterations in the expression levels of ACE and ACE2, at the mRNA and protein levels, in advanced carotid As. These alterations are impacted by sex and PP and may indicate a switch from the balanced RAS/ACE/ACE2 axis in the healthy blood vessel to the unbalanced axis in vascular remodeling due to As.Renin-angiotenzin sistem (RAS) povezan je sa razvojem ateroskleroze (As), uključujući njen nastanak i progresiju. Pored dobro poznatog angiotenzinkonvertujućeg enzima (angiotensin-converting enzyme – ACE), dva nova člana RAS familije povezana su sa remodelovanjem zidova krvnih sudova – ACE2 kao homolog ACE i kolektrin [transmembrane protein 27 (TMEM27)] kao homolog ACE2. Do sada je mali broj studija ispitivao ekspresiju komponenti RAS sistema u tkivu uznapredovalog karotidnog plaka (KP) u odnosu na pol bolesnika i fenotip plaka (FP). Postoje dva tipa KP definisana primenom ultrazvuka – fenotip hipoehogenog plaka (HoP) i fenotip hiperehogenog plaka (HerP). Cilj rada bio je da se ispita da li postoji korelacija između ekspresije komponenti RAS u KP i pola i FP bolesnika. Metode. Ispitano je 74 bolesnika sa uznapredovalim KP koji su bili podvrgnuti operativnoj proceduri karotidne endarterektomije. Ekspresija komponenti RAS sistema u tkivu plaka utvrđena je lančanom reakcijom polimeraze u realnom vremenu (real-time polymerase chain reaction)primenom eseja TaqMan® tehnologije i metode Western blota. Dvosmerna analiza varijanse i Tukey post-hoc test korišćen su za statističku obradu rezultata. Rezultati. Nije utvrđena interakcija FP i pola bolesnika u uticaju na relativnu ekspresiju informacione RNK (iRNK) za ACE i TMEM27 (p > 0,05). U 56,06% uzoraka plaka nije detektovana ekspresija iRNK za ACE2. U plakovima u kojima je detektovana ekspresija iRNK za ACE2, njen nivo bio je viši kod žena sa HoP u poređenju sa ženama sa HerP (p < 0,001). U grupi bolesnika sa fenotipom HoP, žene su imale viši nivo iRNK za ACE2 nego muškarci (p < 0,05). U grupi muškaraca, nivoi ekspresije ACE proteina bili suznačajno niži u fenotipu HoP u poređenju sa HerP (p < 0,001). Žene sa fenotipom HoP imale su značajno viši nivo ACE proteina u poređenju sa muškarcima sa HoP (p < 0,0001). Zaključak. Naši rezultati pokazali su da postoje promene nivoa ekspresije ACE i ACE2, na nivou proteina i iRNK u uznapredovaloj karotidnoj As. Te promene zavise od pola i FP i mogu ukazivati na to da balans ose RAS/ACE/ACE2 koji postoji u zdravom krvnom sudu postaje poremećen tokom remodelovanja zida krvnog suda usled As

Polyphenol-Rich Aronia melanocarpa Juice Consumption Affects LINE-1 DNA Methylation in Peripheral Blood Leukocytes in Dyslipidemic Women

Cardiovascular disease (CVD) is associated with alterations in DNA methylation and polyunsaturated fatty acid (PUFA) profile, both modulated by dietary polyphenols. The present parallel, placebo-controlled study (part of the original clinical study registered as NCT02800967 at www.clinicaltrials.gov) aimed to determine the impact of 4-week daily consumption of polyphenol-rich Aronia melanocarpa juice (AMJ) treatment on Long Interspersed Nucleotide Element-1 (LINE-1) methylation in peripheral blood leukocytes and on plasma PUFAs, in subjects (n = 54, age range of 40.2 ± 6.7 years) at moderate CVD risk, including an increased body mass index, central obesity, high normal blood pressure, and/or dyslipidemia. The goal was also to examine whether factors known to affect DNA methylation (folate intake levels, MTHFR C677T gene variant, anthropometric and metabolic parameters) modulated the LINE-1 methylation levels upon the consumption of polyphenol-rich aronia juice. Experimental analysis of LINE-1 methylation was done by MethyLight method. MTHFR C677T genotypes were determined by the polymerase chain reaction–restriction fragment length polymorphism method, and folate intake was assessed by processing the data from the food frequency questionnaire. PUFAs were measured by gas–liquid chromatography, and serum lipid profile was determined by using Roche Diagnostics kits. The statistical analyses were performed using Statistica software package. In the comparison after vs. before the treatment period, in dyslipidemic women (n = 22), we observed significant decreases in LINE-1 methylation levels (97.54 ± 1.50 vs. 98.39 ± 0.86%, respectively; P = 0.01) and arachidonic acid/eicosapentaenoic acid ratio [29.17 ± 15.21 vs. 38.42 (25.96–89.58), respectively; P = 0.02]. The change (after vs. before treatment) in LINE-1 methylation directly correlated with the presence of MTHFR 677T allele, average daily folate intake, and the change in serum low-density lipoprotein cholesterol but inversely correlated with the change in serum triacylglycerols (R = 0.72, R2 = 0.52, adjusted R2 = 0.36, P = 0.03). The current results imply potential cardioprotective effects of habitual polyphenol-rich aronia juice consumption achieved through the modifications of DNA methylation pattern and PUFAs in subjects at CVD risk, which should be further confirmed. Hence, the precision nutrition-driven modulations of both DNA methylation and PUFA profile may become targets for new approaches in the prevention of CVD

Oestradiol Treatment Counteracts the Effect of Fructose-Rich Diet on Matrix Metalloproteinase 9 Expression and NF kappa B Activation

Fructose-rich diet induces metabolic changes similar to those observed in metabolic syndrome. Among other matrix metalloproteinases, MMP-9 has an important role in adverse cardiac remodelling and might have a role in the development of cardiovascular disorders associated with metabolic syndrome. The changes of MMP-9 expression could be mediated via the NF kappa B pathway. In this study we investigated the effect of fructose-rich diet on MMP-9 expression in the heart of male and female rats, along with the effect of fructose-rich diet and oestradiol on MMP-9 expression in ovariectomized females. We further assessed the effect of fructose-rich diet and oestradiol on NF kappa B activation, measured as the level of p65 phosphorylation at Ser 276. The results showed that the diet regime did not affect the heart mass. Higher MMP-9 gene expression was found in cardiac tissue of male rats fed the fructose-rich diet than in females on the same diet regime. In ovariectomized females, fructose-rich diet upregulated MMP-9 protein and mRNA expression in the heart, as well as phosphorylation of the p65 subunit of NF kappa B at Ser 276. Oestradiol replacement therapy reverted these changes in the heart of ovariectomized females. This study has shown that oestradiol could revert the early molecular changes in MMP-9 expression induced by fructose-rich diet that occurred before cardiac hypertrophy development by decreasing phosphorylation of the NF kappa B p65 subunit at Ser 276

Fructose-rich diet induces gender-specific changes in expression of the renin-angiotensin system in rat heart and upregulates the ACE/AT1R axis in the male rat aorta

Introduction: The cardiovascular renin-angiotensin system (RAS) could be affected by gender and dietary regime. We hypothesized that male rats will be more susceptible to activation of RAS in the heart and aorta, as a response to a fructose-rich diet (FRD). Materials and methods: Both male and female Wistar rats were given a 10% (w/v) fructose solution for 9 weeks. We measured the biochemical parameters, blood pressure (BP) and heart rate. We used Western blot and real-time polymerase chain reaction (PCR) to quantify protein and gene expression. Results: In the male rats, the FRD elevated BP and expression of cardiac angiotensin-converting enzyme (ACE), while the expression of angiotensin-converting enzyme 2 (ACE2) and angiotensin II Type 2 receptor (AT(2)R) were significantly decreased. In female rats, there were no changes in cardiac RAS expression due to FRD. Furthermore, the ACE/AT(1)R axis was overexpressed in the FRD male rats aortae, while only AT(1)R was upregulated in the FRD female rats aortae. ACE2 expression remained unchanged in the aortae of both genders receiving the FRD. Conclusions: The FRD induced gender-specific changes in the expression of the RAS in the heart and aortae of male rats. Further investigations are required in order to get a comprehensive understanding of the underlying mechanisms of gender-specific fructose-induced cardiovascular pathologies

The effect of fructose rich diet on the expression of renin-angiostensin system components and inflamatory molecules in the rat heart: sex specific differences

Ishrana bogata fruktozom predstavlja bitan faktor u razvoju metaboličkog sindroma koji je povezan sa povećanim rizikom za nastanak kardiovaskularnih bolesti. Smatra se da hronična inflamacija ima bitnu ulogu u njegovoj patogenezi. Ženke su zaštićene od ishranom izazvanih metaboličkih poremećaja i hipertenzije u reproduktivnom periodu. Estrogen, koji utiče na normalno funkcionisanje kardiovaskularnog sistema, bi mogao doprinositi uočenim polno specifičnim razlikama u nastanku simptoma metaboličkog sindroma. Proteinske komponente renin-angiotenzin sistema (RAS) imaju bitnu ulogu u nastanku inflamacije, hipertenzije i insulinske rezistencije i na njihovu ekspresiju utiče estradiol. Ova doktorska disertacija je za cilj imala da ispita da li pol i estradiol doprinose promenama u ekspresiji komponenti RAS-a [angiotenzin konvertujućeg enzima (ACE), angiotenzin konvertujućeg enzima 2 (ACE2), angiotenzinskog receptora tipa 1 (AT1R), angiotenzinskog receptora tipa 2 (AT2R) i kolektrina] i medijatora inflamacije i remodelovanja tkiva srca [nuklearnog faktora κB (NFκB), matriks metaloproteinaze-9 (MMP-9) i liganda 16 iz familije hemokina CXC (CXCL16)] kod animalnog modela metaboličkog sindroma. Mužjaci i ženke pacova koji su pili 10% rastvor fruktoze umesto vode u trajanju od 9 nedelja predstavljali su animalni model metaboličkog sindroma korišćen u ovoj studiji. Radi ispitivanja efekata estradiola jedan deo ženki pacova je ovarijektomisan i podeljen u tri grupe pri čemi je jedna grupa pored standardne laboratorijske hrane pila vodu, druga grupa je pila 10% rastvor fruktoze, a treća grupa je pored rasvora fruktoze primala i supstitucionu terapiju estradiolom. Ishrana bogata fruktozom je uzrokovala povećanje krvnog pritiska samo kod mužjaka pacova. Ovaj tip ishrane nije doveo do promena u masi srca niti u odnosu masa srca/masa tela ni kod jednog pola, kao ni kod ovarijektomisanih životinja što ukazuje da se hipertrofija srca nije razvila...Fructose rich diet (FRD) represents an important factor in the development of metabolic syndrome. Metabolic syndrome is associated with increased risk for cardiovascular diseases occurrence and chronic inflammation has a major role in its pathogenesis. Females are protected from diet-induced metabolic disturbances and hypertension in their reproductive period. Estrogen, which influences the cardiovascular system, could contribute to the observed gender-specific differences in the onset of metabolic syndrome. Components of the renin-angiotensin system (RAS), the synthesis of which is mediated by estrogen, have an important role in the inflammatory processes, hypertension and insulin resistance. Almost all of the components of metabolic syndrome increase the activity of RAS, which finally results in increased oxidative stress and inflammation. The aim of this doctoral dissertation was to investigate sex specific changes and role of estradiol in the expression of RAS components [angiotensin converting enzyme (ACE), angiotensin converting enzyme 2 (ACE2), angiotensin receptor type 1 (AT1R), angiotensin receptor type 2 (AT2R) and collectrin], as well as mediators of inflammation and remodeling of heart tissue [nuclear faktor κB (NFκB), matrix metalloproteinse-9 (MMP-9) and CXCL16 chemokine] in the animal model of metabolic syndrome. Male and female rats, which consumed 10% fructose solution instead of water for 9 weeks, represent an animal model of metabolic syndrome that was used in this study. In order to examine the effects of estradiol in the context of FRD, female rats were ovariectomized and divided in three groups: fed normal diet, fed FRD, and fed FRD and subjected to estradiol replacement therapy. FRD increased blood pressure only in male rats. This diet regime didn't cause heart hypertrophy neither in intact males and females, nor in ovariectomized females..

The effect of fructose rich diet on the expression of renin-angiostensin system components and inflamatory molecules in the rat heart: sex specific differences

Ishrana bogata fruktozom predstavlja bitan faktor u razvoju metaboličkog sindroma koji je povezan sa povećanim rizikom za nastanak kardiovaskularnih bolesti. Smatra se da hronična inflamacija ima bitnu ulogu u njegovoj patogenezi. Ženke su zaštićene od ishranom izazvanih metaboličkih poremećaja i hipertenzije u reproduktivnom periodu. Estrogen, koji utiče na normalno funkcionisanje kardiovaskularnog sistema, bi mogao doprinositi uočenim polno specifičnim razlikama u nastanku simptoma metaboličkog sindroma. Proteinske komponente renin-angiotenzin sistema (RAS) imaju bitnu ulogu u nastanku inflamacije, hipertenzije i insulinske rezistencije i na njihovu ekspresiju utiče estradiol. Ova doktorska disertacija je za cilj imala da ispita da li pol i estradiol doprinose promenama u ekspresiji komponenti RAS-a [angiotenzin konvertujućeg enzima (ACE), angiotenzin konvertujućeg enzima 2 (ACE2), angiotenzinskog receptora tipa 1 (AT1R), angiotenzinskog receptora tipa 2 (AT2R) i kolektrina] i medijatora inflamacije i remodelovanja tkiva srca [nuklearnog faktora κB (NFκB), matriks metaloproteinaze-9 (MMP-9) i liganda 16 iz familije hemokina CXC (CXCL16)] kod animalnog modela metaboličkog sindroma. Mužjaci i ženke pacova koji su pili 10% rastvor fruktoze umesto vode u trajanju od 9 nedelja predstavljali su animalni model metaboličkog sindroma korišćen u ovoj studiji. Radi ispitivanja efekata estradiola jedan deo ženki pacova je ovarijektomisan i podeljen u tri grupe pri čemi je jedna grupa pored standardne laboratorijske hrane pila vodu, druga grupa je pila 10% rastvor fruktoze, a treća grupa je pored rasvora fruktoze primala i supstitucionu terapiju estradiolom. Ishrana bogata fruktozom je uzrokovala povećanje krvnog pritiska samo kod mužjaka pacova. Ovaj tip ishrane nije doveo do promena u masi srca niti u odnosu masa srca/masa tela ni kod jednog pola, kao ni kod ovarijektomisanih životinja što ukazuje da se hipertrofija srca nije razvila...Fructose rich diet (FRD) represents an important factor in the development of metabolic syndrome. Metabolic syndrome is associated with increased risk for cardiovascular diseases occurrence and chronic inflammation has a major role in its pathogenesis. Females are protected from diet-induced metabolic disturbances and hypertension in their reproductive period. Estrogen, which influences the cardiovascular system, could contribute to the observed gender-specific differences in the onset of metabolic syndrome. Components of the renin-angiotensin system (RAS), the synthesis of which is mediated by estrogen, have an important role in the inflammatory processes, hypertension and insulin resistance. Almost all of the components of metabolic syndrome increase the activity of RAS, which finally results in increased oxidative stress and inflammation. The aim of this doctoral dissertation was to investigate sex specific changes and role of estradiol in the expression of RAS components [angiotensin converting enzyme (ACE), angiotensin converting enzyme 2 (ACE2), angiotensin receptor type 1 (AT1R), angiotensin receptor type 2 (AT2R) and collectrin], as well as mediators of inflammation and remodeling of heart tissue [nuclear faktor κB (NFκB), matrix metalloproteinse-9 (MMP-9) and CXCL16 chemokine] in the animal model of metabolic syndrome. Male and female rats, which consumed 10% fructose solution instead of water for 9 weeks, represent an animal model of metabolic syndrome that was used in this study. In order to examine the effects of estradiol in the context of FRD, female rats were ovariectomized and divided in three groups: fed normal diet, fed FRD, and fed FRD and subjected to estradiol replacement therapy. FRD increased blood pressure only in male rats. This diet regime didn't cause heart hypertrophy neither in intact males and females, nor in ovariectomized females..

Estradiol Protects Ovariectomized Female Rats Against Fructose Rich Diet Induced Cardiac Inflammation

Congress of the European-Atherosclerosis-Society (EAS), May 29-Jun 01, 2016, Innsbruck, Austri

Fructose-Rich Diet-Induced Changes in the Expression of the Renin Angiotensin System Molecules in the Heart of Ovariectomized Female Rats Could be Reversed by Estradiol

The renin-angiotensin system has been implicated in the development of metabolic syndrome and appears to be a key in the local tissue control of normal cardiac functions. Physiological concentrations of estrogens have been shown to be cardioprotective, especially against the damaging effects of fructose-rich diet. The aim of the study was to investigate the expression of the renin-angiotensin system molecules with potentially deleterious effect on the heart (angiotensin-converting enzyme and angiotensin II type 1 receptor) and those with potentially protective effects, (angiotensin-converting enzyme 2 and angiotensin II type 2 receptor), in ovariectomized fructose fed female rats with 17-estradiol replacement. Real-time PCR and Western blot analysis were used for quantification of gene and protein expression in the heart. Fructose diet increased the expression of angiotensin-converting enzyme and angiotensin II type 1 receptor and decreased the expression of angiotensin-converting enzyme 2 and angiotensin II type 2 receptor. On the other hand, estradiol replacement seems to undo fructose diet effects on cardiac renin-angiotensin system. Downregulation of angiotensin-converting enzyme and angiotensin II type 1 receptor, and reversion of expression of both potentially protective molecules, angiotensin-converting enzyme 2 and angiotensin II type 2 receptor, to the control level in cardiac tissue took place. Obtained results suggest that estradiol may reverse the harmful effect of fructose-rich diet on the expression of renin-angiotensin system molecules. These findings may also be important in further research of phenotypes like insulin resistance, metabolic syndrome, and following cardiovascular pathology in females