Document Detail


Effects of endothelin-1 on differentiation of cardiac myocyte induced from rabbit bone marrow stromal cells.
MedLine Citation:
PMID:  16732986     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
Authors:
Hong-yan Dong; Zhong-ming Zhang; Zhong-xin Zhou
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Chinese medical journal     Volume:  119     ISSN:  0366-6999     ISO Abbreviation:  Chin. Med. J.     Publication Date:  2006 May 
Date Detail:
Created Date:  2006-05-30     Completed Date:  2006-06-29     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7513795     Medline TA:  Chin Med J (Engl)     Country:  China    
Other Details:
Languages:  eng     Pagination:  832-9     Citation Subset:  IM    
Affiliation:
Centre of Neurobiological Research, Xuzhou Medical College, Xuzhou 221002, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Bone Marrow Cells / cytology*,  ultrastructure
Cell Differentiation / drug effects*
Cells, Cultured
Endothelin-1 / pharmacology*
GATA4 Transcription Factor / analysis,  metabolism
Myocytes, Cardiac / cytology*
Myosin Heavy Chains / genetics
Phosphorylation
RNA, Messenger / analysis
Rabbits
Stromal Cells / cytology,  ultrastructure
Chemical
Reg. No./Substance:
0/Endothelin-1; 0/GATA4 Transcription Factor; 0/Myosin Heavy Chains; 0/RNA, Messenger

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