胰高血糖素样肽-2和二肽基肽酶Ⅳ抑制剂联合使用对断奶仔猪肠上皮细胞的影响  

Effects of the Combination of Glucagon-like Peptide-2 and Dipeptidyl Peptidase-ⅣInhibitors on Intestinal Epithelial Cells of Weaned Piglets in vitro

作　　者：贾刚[1] 邓秋红[1] 蒋荣川[1] 赵华[1] 陈小玲[1] 刘光芒[1] 王康宁[1] 

机构地区：[1]四川农业大学动物营养研究所,成都611130

出　　处：《畜牧兽医学报》2017年第1期99-107,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：高等学校博士学科点专项科研基金(博导类)(20125103110011);四川省科技支撑计划(2013NZ0054)

摘　　要：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 conducted to investigate the effects of the combination of Glucagon-like peptide-2（GLP-2）and dipeptidyl peptidase-Ⅳ（DPP-Ⅳ）inhibitors with different concentrations on cell proliferation,metabolism and enzyme activity of primarily intestinal epithelial cells cultured in vitro from 28-day-old weaned piglets,and aimed to explore the method of promoting GLP-2effects.The experiment,taking 28-day-old weaned piglets＇ intestinal epithelial cells as a model,studied the effects of DPP-Ⅳinhibitors（KR62436）with different concentrations on the cell proliferation and metabolism.The 2×3factorial design was adopted to study the effects of combination of GLP-2（1×10^-10,1×10^-9 and 1×10^-8 mol·L^-1）and KR62436（0and 1×10^-11mol·L^-1）on cell proliferation,metabolism and apoptosis.The results were as follows：therewas no significant effect of DPP-Ⅳ with 1×10^-11 mol·L^-1 on cell number,MTT OD,cell protein retention,total cell protein level,LDH activity,CK activity and Na＋,K＋-ATPase activity（P〈0.05）.Compared with the control group,the DPP-Ⅳ group with 1×10^-10 mol·L^-1showed lower MTT OD value（P〈0.05）,and higher LDH and CK activity（P〈0.05）.In the groups that treated with different concentrations of GLP-2and low concentrations of KR62436,with the GLP-2doses increasing,cell number,MTT OD value,cell protein retention,total cell protein level and Na＋,K＋-ATPase activity increased significantly（P〈0.05）,and LDH and CK activity decreased significantly（P〈0.05）.KR62436 could significantly promote GLP-2effects.The results indicate that the low concentration DPP-Ⅳinhibitors have no significant effect on cell growth,while the high concentration DPP-Ⅳinhibitor have a negative effect on cell metabolism and integrity.Combination of GLP-2and low concentrations DPP-Ⅳinhibitors may have favorable effects on intestinal epithelial cell proliferation,metabolism and integrity than GLP-2alone.

关 键 词：胰高血糖素样肽-2 DPP-Ⅳ抑制剂 肠上皮细胞 增殖 完整性 

分 类 号：S828[农业科学—畜牧学] S811.2[农业科学—畜牧兽医]