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

doi:10.11843/j.issn.0366-6964.2017.01.012

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (1): 99-107.doi: 10.11843/j.issn.0366-6964.2017.01.012

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

贾刚^#*，邓秋红^#，蒋荣川，赵华，陈小玲，刘光芒，王康宁

（四川农业大学动物营养研究所，成都 611130）

收稿日期:2016-06-16 出版日期:2017-01-23 发布日期:2017-01-23
通讯作者: 贾刚，E-mail: jiagang700510@163.com
作者简介:贾刚(1970-)，男，四川汉源人，博士，教授，博士生导师，主要从事畜禽肠黏膜发育规律及营养调控研究；邓秋红（1985-），女，四川德阳人，博士生，主要从事畜禽肠黏膜发育规律及营养调控研究，E-mail: dqiuhong818@163.com。二者并列为第一作者
基金资助:
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)

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

JIA Gang^#*, DENG Qiu-hong^#, JIANG Rong-chuan, ZHAO Hua, CHEN Xiao-ling, LIU Guang-mang, WANG Kang-ning

(Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China)

Received:2016-06-16 Online:2017-01-23 Published:2017-01-23

摘要/Abstract

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。

Abstract:

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-2 effects. The experiment, taking 28-day-old weaned piglets’ intestinal epithelial cells as a model, studied the effects of DPP-Ⅳ inhibitors (KR62436) with different concentrations on the cell proliferation and metabolism. The 2×3 factorial 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 (0 and 1×10^-11 mol•L^-1) on cell proliferation, metabolism and apoptosis. The results were as follows: there was 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^-1 showed 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-2 and low concentrations of KR62436, with the GLP-2 doses 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-2 effects. The results indicate that the low concentration DPP-Ⅳ inhibitors have no significant effect on cell growth, while the high concentration DPP-Ⅳ inhibitor have a negative effect on cell metabolism and integrity. Combination of GLP-2 and low concentrations DPP-Ⅳ inhibitors may have favorable effects on intestinal epithelial cell proliferation, metabolism and integrity than GLP-2 alone.

中图分类号:

S828
S811.2

贾刚，邓秋红，蒋荣川，赵华，陈小玲，刘光芒，王康宁. 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[J]. 畜牧兽医学报, 2017, 48(1): 99-107.

JIA Gang, DENG Qiu-hong, JIANG Rong-chuan, ZHAO Hua, CHEN Xiao-ling, LIU Guang-mang, WANG Kang-ning. Effects of the Combination of Glucagon-like Peptide-2 and Dipeptidyl Peptidase-Ⅳ Inhibitors on Intestinal Epithelial Cells of Weaned Piglets in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(1): 99-107.

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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-Ⅳ Inhibitors on Intestinal Epithelial Cells of Weaned Piglets in vitro

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