Project supported by the National Natural Science Foundation of China (No 30171121), by the Key Grant Project of the Chinese Ministry of Education (No 03088), and by the Key Grant Project of the International Joint Key Grant Project of Zhejiang Province (No 2003C24005).
Inducible effects of icariin, icaritin, and desmethylicaritin on directional differentiation of embryonic stem cells into cardiomyocytes in vitro†
Article first published online: 1 APR 2005
Acta Pharmacologica Sinica
Volume 26, Issue 4, pages 477–485, April 2005
How to Cite
ZHU, D.-y. and LOU, Y.-j. (2005), Inducible effects of icariin, icaritin, and desmethylicaritin on directional differentiation of embryonic stem cells into cardiomyocytes in vitro. Acta Pharmacologica Sinica, 26: 477–485. doi: 10.1111/j.1745-7254.2005.00076.x
- Issue published online: 1 APR 2005
- Article first published online: 1 APR 2005
- Received 2004-09-05 Accepted 2005-12-21
- embryonic stem cells;
- directional differentiation;
- α-major histocompatibility complex;
- myosin light chain 2v;
- cell cycle;
Aim: To investigate the possible inducible effects of icariin, icaritin, and desmethylicaritin on the directional differentiation of embryonic stem (ES) cells into cardiomyocytes in vitro. Methods: ES cells were cultivated as embryoid bodies (EBs) in hanging drops with icariin, icaritin, or desmethylicaritin. ES cells treated with retinoic acid and with solvent were used as positive and negative controls, respectively. The cardiomyocytes derived from the ES cells were verified using immunocytochemistry. The expression of cardiac developmentaldependent genes was detected using the reverse transcription-polymerase chain reaction (RT-PCR) method. Cell cycle distribution and apoptosis were analyzed using flow cytometry to determine the partly inducible effect mechanisms involved. Results: The total percentage of beating EBs treated with 1×10−7 mol/L icariin, icaritin, or desmethylicaritin was 87% (P<0.01), 59% (P<0.01), and 49%, respectively. All the beating cardiomyocytes derived from the ES cells expressed cardiac-specific proteins for α-actinin and troponin T. Among them, 1×10−7 mol/L icariin treatment resulted in a significantly advanced and increased mRNA level of α-cardiac major histocompatibility complex (MHC) and myosin light chain 2v (MLC-2v) in EBs in the early cardiac developmental stage. Before shifting to the cardiomyocyte phenotype, icariin could evoke the accumulation of ES cells in G0/G1 and accelerate apoptosis of the cell population (P<0.05). Conclusion: Icariin facilitated the directional differentiation of ES cells into cardiomyocytes at a concentration of 1×10−7 mol/L. The promoting effect of icariin on cardiac differentiation was related to increasing and accelerating gene expression of α-cardiac MHC and MLC-2v, as well as regulating the cell cycles and inducing apoptosis.