Roles of calcium sensing receptor in icariin-induced differentiation of mouse embryonic stem cells to cardiomyocyte
10.3969/j.issn.1000-4718.2016.02.007
- VernacularTitle:C aSR 在淫羊藿苷诱导的胚胎干细胞向心肌细胞分化中的作用
- Author:
Jian SUN
;
Shuzhi BAI
;
Shuang LI
;
Xiaoyi XU
;
Hui YUAN
;
Tao WEI
;
Changqing XU
;
Hairong LUAN
- Publication Type:Journal Article
- Keywords:
Calcium sensing receptors;
Icariin;
Cell differentiation;
Cardiomyocyte
- From:Chinese Journal of Pathophysiology
2016;32(2):234-239
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To study the effect of calcium sensing receptor (CaSR) on icariin (ICA) induced mouse embryonic stem cells ( mESCs) to differentiate into cardiomyocytes in vitro.METHODS:mESCs were cultured to embry-oid bodies ( EBs) by direct suspension method and the differentiation of EBs into cardiomyocytes was induced by ICA.The expression of cardiac specific proteinsα-actinin and cardiac troponin-I ( cTnI) was analyzed by Western blot and immuno-fluorescence.The differentiation rate was determined by flow cytometry.The ultrastructure of the derived cardiomyocytes was further characterized by transmission electron microscopy.The expression of cardiac-specific transcription factors Nkx2.5 and GATA-4,as well as CaSR was detected by Western blot.RESULTS: After induction with ICA, the positive characteristics of myocardial cells appeared in the EBs cultured for 2 d.The expression of cardiac-specific sarcomeric pro-tein actinin (α-actinin) and cTnI showed an overall upward trend by Western blot in different phases of ICA induced differ-entiation.The expression of CaSR, Nkx2.5 and GATA-4 was the highest at an early stage of ICA-induced differentiation. Neomycin (an activator of CaSR) up-regulated CaSR, NKx2.5 and GATA-4 expression in the EBs at early stage of ICA-in-duced differentiation, all of which were reversed by NPS2390 ( an inhibitor of CaSR) .CONCLUSION:CaSR is function-ally expressed in mESC-derived cardiomyocytes, and activation of CaSR is involved in the differentiation of mESCs into car-diomyocytes by facilitating the expression of NKx2.5 and GATA-4.
