摘要
【目的】探讨PDX-1对成肌细胞株C2C12分化为胰岛素分泌细胞的作用。【方法】将C2C12细胞分别经过不同浓度的类胰高血糖素(GLP-1)诱导120h后,瞬间转染胰腺十二指肠同源框(PDX-1)基因,用RT-PCR检测PDX-1的表达,流式细胞仪、放射免疫法检测细胞内和培养基中是否存在胰岛素。【结果】GLP-1可以直接诱导C2C12细胞表达PDX-1;PDX-1瞬时转染C2C12细胞后,胰岛素分泌细胞阳性率比值(1.19±0.10)、培养基中的胰岛素含量(2.45±1.48)×10-6U/mL均明显升高(P<0.05);单独的40nmol/LGLP-1诱导与瞬间转染PDX-1相比,阳性率比值及胰岛素浓度的差别均无统计学意义。【结论】GLP-1可以诱导C2C12表达PDX-1,而且PDX-1可以促进C2C12分泌胰岛素。提示GLP-1诱导C2C12分化为胰岛素分泌细胞可能与PDX-1有关。
[Objective] To investigate the role of PDX-1 in differentiation of myoblast C2C12 cells into insulin-secreting cells. [Methods] C2C12 cells were co-cultivated with various concentration of glucagon-like peptide-1 (GLP-1) for 120 hours. Then the gene of pancreatic-duodenum homeobox-1 (PDX-1) was instantaneously transfected into the C2C12 by plasmid. The amount of PDX-1 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). The insulin in intracellular fluid and supernatant was examined using flow cytometry and radioimmunoassay. [Results] GLP-1 induced the C2C12 expressed PDX-1. The ratio of insulin positive cells percent and insulin in the supernatant in PDX-1(+) cells were (1.19±0.10)×10-6U/mL and (2.45±1.48)μ×10-6U/mL, they were higher than those in PDX-1(-) cells (1.00±0.00)×10-6U/mL and (2.45±1.48)×10-6U/mL significantly (P< 0.05), but that in the cells, which were co-cultivated with 40 nmol/L GLP-1 and that in the PDX-1(+) cells were not different (P >0.05). [Conclusion] GLP-1 could induce the C2C12 expressed PDX-1 and PDX-1 could make the C2C12 cells secrete insulin. So the PDX-1 expressing was one of the reasons that GLP-1 induces differentiation of C2C12 cells into insulin-secreting cells.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2005年第4期409-412,共4页
Journal of Sun Yat-Sen University:Medical Sciences
基金
教育部博士学科点专项科研基金资助项目(20020558077)
中国博士后基金资助项目
