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Effects of endothelin-1 on differentiation of cardiac myocyte induced from rabbit bone marrow stromal cells / 中华医学杂志(英文版)
Chinese Medical Journal ; (24): 832-839, 2006.
Article in English | WPRIM | ID: wpr-265294
ABSTRACT
<p><b>BACKGROUND</b>Cardiomyocyte transplantation for the therapy of myocardial ischaemia is being paid close attention. However, how the microenvironment controls the differentiation of transplanted bone marrow stromal cells (BMSCs) is unknown. Endothelin-1 (ET-1), a cytokine, increases during myocardial infarction, but it is not known whether ET-1 is responsible for the fate of transplanted BMSCs. In the present study, we investigated the effects of ET-1 on differentiation and maturation of induced rabbit BMSCs, in vitro, to elucidate the cellular biological mechanisms.</p><p><b>METHODS</b>The proliferation of BMSCs isolated from femur of rabbits was induced by ET-1 only, by 5-azacytidine (5-aza) or ET-1 combined with 5-aza. After 4 weeks of induced culturing, the differentiation rate and the diameter of induced myocyte like cells were estimated and the expressions of GATA-4 protein and phosphorylation level were assayed by Western-blot, RT-PCR analysis of beta-myosin heavy chain (MHC). mRNA expression, levels of troponin-I by immunohistochemical staining and ultrastructure of induce-cultured BMSCs were also determined.</p><p><b>RESULTS</b>By induction with ET-1 and 5-aza, mean cell diameter of induced BMSCs was larger than induced with 5-aza [(6.26 +/- 0.22) microm cf (5.29 +/- 0.19) microm] (P < 0.001). There was no difference in rate of differentiation of myocyte like cells between the groups induced with 5-aza and ET-1 combined with 5-aza [(29.82 +/- 0.23)% cf (29.94 +/- 0.18)%] (P > 0.05). The expressions of GATA-4 protein and phosphorylation were enhanced significantly in groups induced with ET-1 combined with 5-aza (P < 0.05). In the group induced with ET-1 combined with 5-aza, expression of beta-MHC mRNA was higher than control [(0.122 +/- 0.008) cf (0.022 +/- 0.003)] (P < 0.01), and more troponin-I positive cells were also detected in this group. Differentiated BMSCs showed formations of myofilaments and primitive sarcomere, i.e., morphological characteristics of myocyte like cells.</p><p><b>CONCLUSIONS</b>This study suggests that induced culturing of BMSCs by ET-1 combined with 5-aza can express cardiomyocytic characteristics whereas ET-1 alone could not induce BMSCs to differentiate to myocyte like cells. ET-1 upregulates the expression of GATA-4 protein and phosphorylation level of induced BMSCs, and rapidly promotes the differentiation and maturation of myocyte like cells from BMSCs.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Pharmacology / Phosphorylation / RNA, Messenger / Bone Marrow Cells / Cell Differentiation / Cells, Cultured / Stromal Cells / Myosin Heavy Chains / Endothelin-1 / Cell Biology Limits: Animals Language: English Journal: Chinese Medical Journal Year: 2006 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Pharmacology / Phosphorylation / RNA, Messenger / Bone Marrow Cells / Cell Differentiation / Cells, Cultured / Stromal Cells / Myosin Heavy Chains / Endothelin-1 / Cell Biology Limits: Animals Language: English Journal: Chinese Medical Journal Year: 2006 Type: Article