College of Medicine, CHA University, CHA General Hospital, Seoul 135-081, Korea.
Biol Chem. 2010 Sep;391(9):1019-29. doi: 10.1515/BC.2010.101.
The deterioration of β cells in the pancreas is a crucial factor in the progression of diabetes mellitus; therefore, the recovery of β cells is of vital importance for effective diabetic therapeutic strategies. Partially pancreatectomized rats have been used for the investigation of pancreatic regeneration. Because it was determined that tissue extract from the partially-dissected pancreas induces pancreatic differentiation in embryonic stem cells, paracrine factors were thought to be involved in the regeneration. In this study, we screened for genes that had higher mRNA levels 2 days after 60%-pancreatectomy. The genes were isolated using subtractive hybridization and DNA sequencing. Twelve genes (adipsin, Aplp2, Clu, Col1a2, Glul, Krt8, Lgmn, LOC299907, LOC502894, Pla2g1b, Reg3α and Xbp1) were identified, and RT-PCR and real-time PCR analyses were performed to validate their expression levels. Among the genes identified, three genes (Glul, Lgmn and Reg3a) were selected for further analyses. Assays revealed that Glul and Reg3α enhance cell growth. Glul, Lgmn and Reg3α change the expression level of islet marker genes, where NEUROD, NKX2.2, PAX4 and PAX6 are up-regulated and somatostatin is down-regulated. Thus, we believe that Glul, Lgmn and Reg3a can serve as novel targets in diabetes mellitus genetic therapy.
胰岛β细胞的衰退是糖尿病进展的一个关键因素；因此，β细胞的恢复对于有效的糖尿病治疗策略至关重要。部分胰腺切除术大鼠已被用于研究胰腺再生。因为已经确定部分切除的胰腺组织提取物可在胚胎干细胞中诱导胰腺分化，所以认为旁分泌因子参与了再生过程。在这项研究中，我们筛选了在 60%胰腺切除后 2 天 mRNA 水平较高的基因。使用消减杂交和 DNA 测序分离基因。鉴定出 12 个基因（adipsin、Aplp2、Clu、Col1a2、Glul、Krt8、Lgmn、LOC299907、LOC502894、Pla2g1b、Reg3α 和 Xbp1），并进行 RT-PCR 和实时 PCR 分析以验证其表达水平。在所鉴定的基因中，选择了三个基因（Glul、Lgmn 和 Reg3a）进行进一步分析。检测结果表明，Glul 和 Reg3α 可增强细胞生长。Glul、Lgmn 和 Reg3α 改变胰岛标记基因的表达水平，其中 NEUROD、NKX2.2、PAX4 和 PAX6 上调，生长抑素下调。因此，我们认为 Glul、Lgmn 和 Reg3a 可以作为糖尿病遗传治疗的新靶点。
