Study On The Mechanism Of SGLT1 And GLUT5 Regulated By Transcription Factor GATA3 In Chicken Intestine

Dietary carbohydrate digestion and absorption played an important role in the animal normal growth and development. Carbohydrate was one of the major necessary nutrients in animal growth. Carbohydrate was absorbed in the small intestine. Sugars were transported by monosaccharide transport carrier protein SGLT1 and GLUT5 in the brush border membrane (BBM) of intestinal epithelial cells. So regulating the expression of two genes could affect absorption of sugar and growth and development of animal. Transcription factor GATA3 was a member of GATA family. It played essential roles in controlling cell fates and regulate the expression of the genes in many tissue cells. This study explored transcription factor GATA3 regulated expression mechanism of two kinds of monosaccharide transport carrier proteins. Frist to screen out the best conditions and nutrition factors (inoculation density and serum type) about primary cultivation of chicken small intestinal epithelial cells, then we cultivated and identified cells. The information of GATA3 was analysed to provide the reference 5’ upstream GATA3 binding sites of SGLT1 and GLUTS. The GATA3 was transfected into chicken intestinal epithelial cells by pcDNA3.1. The absolute expression of target genes GATA3, SGLT1 and GLUT5 in chicken intestinal epithelial cells were tested by Real-time PCR.Results:with 6.5×105/mL as the initial seeding density, chicken intestinal epithelial cells showed the best growth state. Compared with culture medium added to fetal bovine serum, culture medium added to chicken serum was more suitable for the growth of chicken intestinal epithelial cells in vitro. The chicken intestinal epithelial cells were cultivated and identified successfully. Upstream of SGLT1 and GLUT5 contained many GATA3 binding sites. Transfected group compared with blank control group and nagetive control group, the expression of GATA3 was highly significant difference(P<0.01), this showed the eukaryotic expression vector pcDNA3.1-GATA3 was transfected into chicken intestinal epithelial cells successfully by liposome transfection method. After overexpression of GATA3 in chicken intestinal epithelial cells, transfected group compared with blank control group and nagetive control group, the expression of GLUT5 was highly significant difference(P<0.01), but the expression of SGLT1 was not significant difference(P>0.05).Conclusion:the increase of GATA3 may help promote the increase of GLUT5 in intestine, but it has no effect on the expression of SGLT1. It provided a theoretical basis for improving nutrition absorption of dietary carbohydrate.