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Therapeutic Effect of Iron Citrate in Blocking Calcium Deposition in High Pi-Calcified VSMC: Role of Autophagy and Apoptosis

Identifieur interne : 000328 ( Main/Exploration ); précédent : 000327; suivant : 000329

Therapeutic Effect of Iron Citrate in Blocking Calcium Deposition in High Pi-Calcified VSMC: Role of Autophagy and Apoptosis

Auteurs : Paola Ciceri ; Monica Falleni ; Delfina Tosi ; Carla Martinelli ; Stefania Cannizzo ; Giulia Marchetti ; Antonella D Rminio Monforte ; Gaetano Bulfamante ; Geoffrey A. Block ; Piergiorgio Messa ; Mario Cozzolino [Italie]

Source :

RBID : PMC:6928632

Abstract

In chronic kidney disease (CKD), the first cause of mortality is cardiovascular disease induced mainly by vascular calcification (VC). Recently, iron-based phosphate binders have been proposed in advanced CKD to treat hyperphosphatemia. We studied the effect of iron citrate (iron) on the progression of calcification in high-phosphate (Pi) calcified VSMC. Iron arrested further calcification when added on days 7–15 in the presence of high Pi (1.30 ± 0.03 vs 0.61 ± 0.02; OD/mg protein; day 15; Pi vs Pi + Fe, p < 0.01). We next investigated apoptosis and autophagy. Adding iron to high-Pi-treated VSMC, on days 7–11, decreased apoptotic cell number (17.3 ± 2.6 vs 11.6 ± 1.6; Annexin V; % positive cells; day 11; Pi vs Pi + Fe; p < 0.05). The result was confirmed thorough analysis of apoptotic nuclei both in VSMCs and aortic rings treated on days 7–15 (3.8 ± 0.2 vs 2.3 ± 0.3 and 4.0 ± 0.3 vs 2.2 ± 0.2; apoptotic nuclei; arbitrary score; day 15; Pi vs Pi + Fe; VSMCs and aortic rings; p < 0.05). Studying the prosurvival axis GAS6/AXL, we found that iron treatment on days 9–14 counteracted protein high-Pi-stimulated down-regulation and induced its de novo synthesis. Moreover, iron added on days 9–15 potentiated autophagy, as detected by an increased number of autophagosomes with damaged mitochondria and an increase in autophagic flux. Highlighting the effect of iron on apoptosis, we demonstrated its action in blocking the H2O2-induced increase in calcification added both before high Pi treatment and when the calcification was already exacerbated. In conclusion, we demonstrate that iron arrests further high Pi-induced calcium deposition through an anti-apoptotic action and the induction of autophagy on established calcified VSMC.


Url:
DOI: 10.3390/ijms20235925
PubMed: 31775364
PubMed Central: 6928632


Affiliations:

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Le document en format XML

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<email>giulia.marchetti@unimi.it</email>
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<name sortKey="D Rminio Monforte, Antonella" sort="D Rminio Monforte, Antonella" uniqKey="D Rminio Monforte A" first="Antonella" last="D Rminio Monforte">Antonella D Rminio Monforte</name>
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<nlm:aff id="af3-ijms-20-05925">Department of Health Sciences, Clinic of Infectious Diseases, University of Milan, ASST Santi Paolo e Carlo, 20142 Milan, Italy;
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<name sortKey="Bulfamante, Gaetano" sort="Bulfamante, Gaetano" uniqKey="Bulfamante G" first="Gaetano" last="Bulfamante">Gaetano Bulfamante</name>
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<nlm:aff id="af2-ijms-20-05925">Department of Health Sciences, Division of Pathology, University of Milan, ASST Santi Paolo e Carlo, 20142 Milan, Italy;
<email>monica.falleni@unimi.it</email>
(M.F.);
<email>delfina.tosi@unimi.it</email>
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<email>carla.martinelli@unimi.it</email>
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<name sortKey="Block, Geoffrey A" sort="Block, Geoffrey A" uniqKey="Block G" first="Geoffrey A" last="Block">Geoffrey A. Block</name>
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<nlm:aff id="af4-ijms-20-05925">Nephrology, Denver Nephrolgysts PC, Denver, CO 80230, USA;
<email>geoff.block@reatapharma.com</email>
</nlm:aff>
</affiliation>
</author>
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<name sortKey="Messa, Piergiorgio" sort="Messa, Piergiorgio" uniqKey="Messa P" first="Piergiorgio" last="Messa">Piergiorgio Messa</name>
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<nlm:aff id="af1-ijms-20-05925">Renal Research Laboratory, Department of Nephrology, Dialysis and Renal Transplant, Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico and Fondazione D’Amico per la Ricerca sulle Malattie Renali, 20100 Milan, Italy;
<email>p.ciceri@hotmail.it</email>
(P.C.);
<email>piergiorgio.messa@unimi.it</email>
(P.M.)</nlm:aff>
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<p>In chronic kidney disease (CKD), the first cause of mortality is cardiovascular disease induced mainly by vascular calcification (VC). Recently, iron-based phosphate binders have been proposed in advanced CKD to treat hyperphosphatemia. We studied the effect of iron citrate (iron) on the progression of calcification in high-phosphate (Pi) calcified VSMC. Iron arrested further calcification when added on days 7–15 in the presence of high Pi (1.30 ± 0.03 vs 0.61 ± 0.02; OD/mg protein; day 15; Pi vs Pi + Fe,
<italic>p</italic>
< 0.01). We next investigated apoptosis and autophagy. Adding iron to high-Pi-treated VSMC, on days 7–11, decreased apoptotic cell number (17.3 ± 2.6 vs 11.6 ± 1.6; Annexin V; % positive cells; day 11; Pi vs Pi + Fe;
<italic>p</italic>
< 0.05). The result was confirmed thorough analysis of apoptotic nuclei both in VSMCs and aortic rings treated on days 7–15 (3.8 ± 0.2 vs 2.3 ± 0.3 and 4.0 ± 0.3 vs 2.2 ± 0.2; apoptotic nuclei; arbitrary score; day 15; Pi vs Pi + Fe; VSMCs and aortic rings;
<italic>p</italic>
< 0.05). Studying the prosurvival axis GAS6/AXL, we found that iron treatment on days 9–14 counteracted protein high-Pi-stimulated down-regulation and induced its de novo synthesis. Moreover, iron added on days 9–15 potentiated autophagy, as detected by an increased number of autophagosomes with damaged mitochondria and an increase in autophagic flux. Highlighting the effect of iron on apoptosis, we demonstrated its action in blocking the H
<sub>2</sub>
O
<sub>2</sub>
-induced increase in calcification added both before high Pi treatment and when the calcification was already exacerbated. In conclusion, we demonstrate that iron arrests further high Pi-induced calcium deposition through an anti-apoptotic action and the induction of autophagy on established calcified VSMC.</p>
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<name sortKey="Cozzolino, M" uniqKey="Cozzolino M">M. Cozzolino</name>
</author>
<author>
<name sortKey="Ciceri, P" uniqKey="Ciceri P">P. Ciceri</name>
</author>
<author>
<name sortKey="Volpi, E M" uniqKey="Volpi E">E.M. Volpi</name>
</author>
<author>
<name sortKey="Olivi, L" uniqKey="Olivi L">L. Olivi</name>
</author>
<author>
<name sortKey="Messa, P G" uniqKey="Messa P">P.G. Messa</name>
</author>
</analytic>
</biblStruct>
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<analytic>
<author>
<name sortKey="Darzynkiewicz, Z" uniqKey="Darzynkiewicz Z">Z. Darzynkiewicz</name>
</author>
<author>
<name sortKey="Juan, G" uniqKey="Juan G">G. Juan</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Son, B K" uniqKey="Son B">B.K. Son</name>
</author>
<author>
<name sortKey="Kozaki, K" uniqKey="Kozaki K">K. Kozaki</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Proudfoot, D" uniqKey="Proudfoot D">D. Proudfoot</name>
</author>
<author>
<name sortKey="Skepper, J N" uniqKey="Skepper J">J.N. Skepper</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Ewence, A E" uniqKey="Ewence A">A.E. Ewence</name>
</author>
<author>
<name sortKey="Bootman, M" uniqKey="Bootman M">M. Bootman</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Levine, B" uniqKey="Levine B">B. Levine</name>
</author>
<author>
<name sortKey="Klionsky, D J" uniqKey="Klionsky D">D.J. Klionsky</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Kim, K W" uniqKey="Kim K">K.W. Kim</name>
</author>
<author>
<name sortKey="Hwang, M" uniqKey="Hwang M">M. Hwang</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Green, D R" uniqKey="Green D">D.R. Green</name>
</author>
<author>
<name sortKey="Galluzzi, L" uniqKey="Galluzzi L">L. Galluzzi</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Dai, X Y" uniqKey="Dai X">X.Y. Dai</name>
</author>
<author>
<name sortKey="Zhao, M M" uniqKey="Zhao M">M.M. Zhao</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Filomeni, G" uniqKey="Filomeni G">G. Filomeni</name>
</author>
<author>
<name sortKey="De Zio, D" uniqKey="De Zio D">D. De Zio</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Zarjou, A" uniqKey="Zarjou A">A. Zarjou</name>
</author>
<author>
<name sortKey="Jeney, V" uniqKey="Jeney V">V. Jeney</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Sikura, K E" uniqKey="Sikura K">K.É. Sikura</name>
</author>
<author>
<name sortKey="Potor, L" uniqKey="Potor L">L. Potor</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Berberat, P O" uniqKey="Berberat P">P.O. Berberat</name>
</author>
<author>
<name sortKey="Katori, M" uniqKey="Katori M">M. Katori</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Ciceri, P" uniqKey="Ciceri P">P. Ciceri</name>
</author>
<author>
<name sortKey="Elli, F" uniqKey="Elli F">F. Elli</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Ciceri, P" uniqKey="Ciceri P">P. Ciceri</name>
</author>
<author>
<name sortKey="Volpi, E" uniqKey="Volpi E">E. Volpi</name>
</author>
</analytic>
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</listBibl>
</div1>
</back>
</TEI>
<affiliations>
<list>
<country>
<li>Italie</li>
</country>
<region>
<li>Lombardie</li>
</region>
<settlement>
<li>Milan</li>
</settlement>
</list>
<tree>
<noCountry>
<name sortKey="Block, Geoffrey A" sort="Block, Geoffrey A" uniqKey="Block G" first="Geoffrey A" last="Block">Geoffrey A. Block</name>
<name sortKey="Bulfamante, Gaetano" sort="Bulfamante, Gaetano" uniqKey="Bulfamante G" first="Gaetano" last="Bulfamante">Gaetano Bulfamante</name>
<name sortKey="Cannizzo, Stefania" sort="Cannizzo, Stefania" uniqKey="Cannizzo S" first="Stefania" last="Cannizzo">Stefania Cannizzo</name>
<name sortKey="Ciceri, Paola" sort="Ciceri, Paola" uniqKey="Ciceri P" first="Paola" last="Ciceri">Paola Ciceri</name>
<name sortKey="D Rminio Monforte, Antonella" sort="D Rminio Monforte, Antonella" uniqKey="D Rminio Monforte A" first="Antonella" last="D Rminio Monforte">Antonella D Rminio Monforte</name>
<name sortKey="Falleni, Monica" sort="Falleni, Monica" uniqKey="Falleni M" first="Monica" last="Falleni">Monica Falleni</name>
<name sortKey="Marchetti, Giulia" sort="Marchetti, Giulia" uniqKey="Marchetti G" first="Giulia" last="Marchetti">Giulia Marchetti</name>
<name sortKey="Martinelli, Carla" sort="Martinelli, Carla" uniqKey="Martinelli C" first="Carla" last="Martinelli">Carla Martinelli</name>
<name sortKey="Messa, Piergiorgio" sort="Messa, Piergiorgio" uniqKey="Messa P" first="Piergiorgio" last="Messa">Piergiorgio Messa</name>
<name sortKey="Tosi, Delfina" sort="Tosi, Delfina" uniqKey="Tosi D" first="Delfina" last="Tosi">Delfina Tosi</name>
</noCountry>
<country name="Italie">
<region name="Lombardie">
<name sortKey="Cozzolino, Mario" sort="Cozzolino, Mario" uniqKey="Cozzolino M" first="Mario" last="Cozzolino">Mario Cozzolino</name>
</region>
</country>
</tree>
</affiliations>
</record>

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