Influence of retinoic acid on TBX1 expression in myocardial cells induced by Shh and Fgf8

doi:10.1007/s11684-009-0007-8

Front Med Chin

2009, Vol. 3

Issue (1) : 61-66 https://doi.org/10.1007/s11684-009-0007-8

RESEARCH ARTCILE

Influence of retinoic acid on TBX1 expression in myocardial cells induced by Shh and Fgf8

Miao LIU, Xiaoyan WU, Jiawei XU, Runming JIN(

)

Department of Paediatrics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China

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Abstract

The aim of this study was to explore the regulatory mechanism of retinoic acid (RA) on the TBX1 gene expression in myocardial cells. Ventricular cardiocytes were isolated from neonatal rats and cultured, and then treated with different concentrations of retinoic acid. The expression of Shh and Fgf8 at mRNA and protein levels in neonatal rat myocardial cells were measured by using RT-PCR and Western blot technique, respectively. There was basal expression of Shh and Fgf8 in the control group. When treated with 3×10^-7 mol/L RA, we observed that the expression of Shh mRNA and protein in neonatal rat myocardial cells were up-regulated by 1.51 (P<0.05) and 1.10 times (P<0.05), respectively. In comparison with the control group, under the concentration of 5×10^-7 mol/L RA, they were up-regulated by 2.21 (P<0.05) and 2.38 times (P<0.05) individually. Meanwhile, we could detect that the expression of Fgf8 mRNA and protein were up-regulated by 2.50 times (P<0.05) and 80% (P<0.05) separately compared with the control group after stimulation of 3×10^-7 mol/L RA, and they were up-regulated by 3.48 (P<0.05) and 2.04 times (P<0.05) individually after stimulation of 5×10^-7 mol/L RA. The results indicated that RA could induce the expression of Shh and Fgf8 in neonatal rat myocardial cells. At the same time, it has shown that Shh and Fgf8 were involved in the regulation process of RA on TBX1 expression.

Keywords retinoic acid Tbx1 protein Shh protein Fgf8 protein

Corresponding Author(s): JIN Runming,Email:jinrunm@public.wh.hb.cn

Issue Date: 05 March 2009

Cite this article:

Miao LIU,Xiaoyan WU,Jiawei XU, et al. Influence of retinoic acid on TBX1 expression in myocardial cells induced by Shh and Fgf8[J]. Front Med Chin, 2009, 3(1): 61-66.

URL:

https://academic.hep.com.cn/fmd/EN/10.1007/s11684-009-0007-8
https://academic.hep.com.cn/fmd/EN/Y2009/V3/I1/61

Tab.1 The primer sequence and amplification conditions of genes

Fig.1 The cardiomyocytes in culture (× 200)

Fig.2 Purification of cardiomyocytes detected by FACS. M1 represents negative cells; M2 represents positive cells.

Fig.3 Expression of Shh and Fgf8 mRNA in cultured cardiomyocytes by RT-PCR. The relative level of Shh and Fgf8 were determined by quantification of RT-PCR bands and were normalized against the level of all-actin. M: marker; 1: Shh mRNA in cultured cells; 2: Shh mRNA in cultured cells added with 0.1% DMSO; 3: Shh mRNA in cultured cells added with 0.1% DMSO and 1 × 10mol/L RA; 4: Shh mRNA in cultured cells added with 0.1% DMSO and 3 × 10 mol/L RA; 5: Shh mRNA in cultured cells added with 0.1% DMSO and 5×10 mol/L RA; 6: Fgf8 mRNA in cultured cells; 7: Fgf8 mRNA in cultured cells added with 0.1% DMSO; 8: Fgf8 mRNA in cultured cells added with 0.1% DMSO and 1 × 10 mol/L RA; 9: Fgf8 mRNA in cultured cells added with 0.1% DMSO and 3 × 10 mol/L RA; 10: Fgf8 mRNA in cultured cells added with 0.1%DMSO and 5×10 mol/L RA. RA: retinoic acid; DMSO: dimethyl sulphoxide.

Fig.4 Expression of Shh and Fgf8 protein in cardiomyocytes by Western blot. The relative level of Shh and Fgf8 were determined by quantification of Western blot bands and were normalized against the level of GAPDH. M: marker; 1: cultured cells; 2: cultured cells added with 0.1% DMSO; 3: cultured cells added with 0.1% DMSO and 1 × 10 mol/L RA; 4: cultured cells added with 0.1% DMSO and 3 × 10 mol/L RA; 5: cultured cells added with 0.1% DMSO and 5 × 10 mol/L RA. RA: retinoic acid; DMSO: dimethyl sulphoxide.

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