生物技术通报 ›› 2021, Vol. 37 ›› Issue (2): 246-252.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0713

• 技术与方法 • 上一篇    下一篇

猪十二指肠组织灌流培养与静态培养比较

王绿阳1,2(), 康翠翠1,2, 冯江银1,2, 丁立人2, 杭苏琴1,2()   

  1. 1.南京农业大学国家动物消化道营养国际联合研究中心 江苏省消化道营养与动物健康重点实验室 消化道微生物研究室,南京 210095
    2.南京农业大学动物科学类国家级实验教学示范中心,南京 210095
  • 收稿日期:2020-06-09 出版日期:2021-02-26 发布日期:2021-02-26
  • 作者简介:王绿阳,女,硕士研究生,研究方向:猪胃肠营养感应;E-mail: 2018105079@njau.edu.cn
  • 基金资助:
    国家重点基础研究发展计划(973项目)(2013CB127301)

Comparative Study of Pig Duodenum Between Tissue Perfusion Culture and Static Culture

WANG Lü-yang1,2(), KANG Cui-cui1,2, FENG Jiang-yin1,2, DING Li-ren2, HANG Su-qin1,2()   

  1. 1. National Center for International Research on Animal Gut Nutrition/Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health/Laboratory of Gastrointestinal Microbiology,Nanjing Agricultural University,Nanjing 210095
    2. National Experimental Teaching Demonstration Center of Animal Science,Nanjing Agricultural University,Nanjing 210095
  • Received:2020-06-09 Published:2021-02-26 Online:2021-02-26

摘要:

组织灌流培养和静态培养是肠道组织体外培养最常见的两种方法,但目前尚不清楚这两种培养方法哪种更适合用于体外胃肠激素分泌的研究。以猪十二指肠组织为研究对象,首先比较L-精氨酸下两种体外培养方法对组织胆囊收缩素(Cholecystokinin,CCK)分泌和组织活性乳酸脱氢酶(Lactate dehydrogenase,LDH)分泌的影响;并进一步与体内十二指肠L-精氨酸灌注对血液CCK的变化水平进行比较,确定更适合用于体外胃肠激素分泌研究的方法。结果表明,组织灌流培养下培养液CCK浓度随时间增长呈平稳波动,LDH浓度随时间增长而逐渐降低;而静态培养下CCK浓度随时间增长呈累积效应,LDH浓度则随时间增长而逐渐升高。此外,组织灌流培养CCK的分泌过程与血液CCK的变化非常相似。以上结果表明组织灌流培养更适用于体外胃肠激素分泌的研究。

关键词: 组织灌流培养, 组织静态培养, L-精氨酸, 胆囊收缩素, 乳酸脱氢酶

Abstract:

Tissue perfusion culture and static culture are the two most common methods of intestinal tissue culture in vitro,however,it is unclear which method is more suitable for the study of gut hormone secretion. This research used pig duodenum as the research object and chose L-arginine as stimulus. Firstly,we compared the effects of the 2 in vitro methods on cholecystokinin and lactate dehydrogenase secretion stimulated with L-arginine;furthermore,we made a comparison of the CCK levels in the duodenum treated with L-arginine in vivo,aiming to determine which method was more suitable for the study of gut hormone secretion in vitro. Results showed that CCK concentration fluctuated steadily and LDH concentration decreased gradually with the increase of time in the tissue perfusion culture;while in static culture,CCK and LDH concentration increased over time. In addition,the secretion of CCK in the tissue perfusion culture was very similar to that in blood. Above results demonstrate that the tissue perfusion culture is more suitable for the study of gut hormone secretion in vitro.

Key words: tissue perfusion culture, tissue static culture, L-arginine, CCK, LDH