论文部分内容阅读
本试验旨在研究不同浓度的胰高血糖素样肽-2（Glucagon-like peptide 2，GLP-2）和二肽基肽酶Ⅳ（Dipeptidyl peptidase-Ⅳ，DPP-Ⅳ）抑制剂联合使用对体外原代培养的28日龄断奶仔猪肠上皮细胞增殖、代谢及相关酶活力的影响，初步探讨提高GLP-2作用效果的方法。试验首先采用单因子试验设计探讨不同浓度DPP-Ⅳ抑制剂（KR62436）对断奶仔猪肠黏膜上皮细胞增殖及代谢的影响；然后采用2×3因子设计，考察添加不同浓度的GLP-2（1×10-10、1×10-9、1×10-8 mol?L-1）和KR62436（0和1×10-11 mol?L-1）对细胞增殖、代谢及凋亡的影响。结果发现，培养液中单独添加1×10-11 mol?L-1 KR62436，细胞数量、MTT OD值、细胞沉积蛋白量、细胞总蛋白量、胞外LDH、CK和Na+,K+-ATP酶活均无显著变化（P>0.05）；当KR62436浓度升高到1×10-10 mol?L-1，细胞MTT OD值显著低于对照组（P<0.05），胞外LDH和CK活力显著高于对照组（P<0.05）。培养液中同时添加GLP-2和低剂量的KR62436，随着培养液中GLP-2浓度的增加，细胞数量、MTT OD值、细胞蛋白沉积量、细胞总蛋白含量和Na+,K+-ATP酶活力显著增加（P<0.05），胞外LDH、CK活力显著降低（P<0.05）。KR62436显著提高了GLP-2的作用效果。结果表明，低剂量KR62436对细胞生长的影响不显著，高剂量的KR62436对细胞的代谢和完整性有负面效应；但低剂量的KR62436能够促进GLP-2对细胞增殖、代谢和细胞完整性的作用效果。
This study was designed to investigate the effects of different concentrations of Glucagon-like peptide 2 (GLP-2) and Dipeptidyl peptidase-IV (DPP-IV) Primary culture of 28-day-old piglets weaned pig intestinal epithelial cells proliferation, metabolism and related enzyme activity, preliminary study to improve the effect of GLP-2 method. In the first experiment, the effect of different concentrations of DPP-IV inhibitor (KR62436) on the proliferation and metabolism of intestinal mucosal epithelial cells in weaned piglets was investigated by single factor experiment design. Then, 2 × 3 factorial design was used to investigate the effects of adding different concentrations of GLP- 10-10, 1 × 10-9, 1 × 10-8 mol·L-1) and KR62436 (0 and 1 × 10-11 mol·L-1) on cell proliferation, metabolism and apoptosis. The results showed that 1 × 10-11 mol·L-1 KR62436, cell number, MTT OD, cell sediment protein, total cellular protein, extracellular LDH, CK and Na +, K + -ATP activity (P> 0.05). When the concentration of KR62436 was increased to 1 × 10-10 mol·L-1, the MTT OD of cells was significantly lower than that of the control group (P <0.05), and the activities of extracellular LDH and CK were significantly increased In the control group (P & lt; 0.05). With the addition of GLP-2 and low-dose KR62436 in culture medium, the cell number, MTT OD value, cell protein deposition, total cellular protein content and Na +, K + -ATPase activity (P <0.05). The activities of extracellular LDH and CK were significantly decreased (P <0.05). KR62436 significantly increased the effect of GLP-2. The results showed that low doses of KR62436 had no significant effect on cell growth. High doses of KR62436 had a negative effect on cell metabolism and integrity; however, low doses of KR62436 promoted the effect of GLP-2 on cell proliferation, metabolism and cell integrity effect.