猪十二指肠组织灌流培养与静态培养比较

doi:10.13560/j.cnki.biotech.bull.1985.2020-0713

摘要/Abstract

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

王绿阳, 康翠翠, 冯江银, 丁立人, 杭苏琴. 猪十二指肠组织灌流培养与静态培养比较[J]. 生物技术通报, 2021, 37(2): 246-252.

WANG Lü-yang, KANG Cui-cui, FENG Jiang-yin, DING Li-ren, HANG Su-qin. Comparative Study of Pig Duodenum Between Tissue Perfusion Culture and Static Culture[J]. Biotechnology Bulletin, 2021, 37(2): 246-252.

图/表 6

图1 体内十二指肠灌注实验程序图

图2 组织灌流培养对CCK分泌的影响 A：组织块完整培养下CCK的分泌;B：组织块剪碎培养下CCK的分泌;C：不同组织形态培养下CCK分泌的比较（*P<0.05,表示10,20,50 mmol/L L-Arg vs 0 mmol/L L-Arg;# P<0.05,表示组织机械破碎培养 vs 组织块培养,n=4）

图3 组织静态培养对CCK分泌的影响 A：组织块培养下CCK的分泌;B：组织机械破碎培养下CCK的分泌;C：不同组织形态培养下CCK分泌的比较（*P<0.05,表示10,20,50 mmol/L L-Arg vs 0 mmol/L L-Arg;# P<0.05,表示组织剪碎培养 vs 组织块培养,n=6）

图4 不同培养方式下CCK的分泌过程 * P<0.05,灌流培养;# P<0.05,表示静态培养,n=6

图5 不同培养方式下LDH分泌的动态过程 * P<0.05,灌流培养;# P<0.05,静态培养,n=6

图6 体内灌流实验中血浆CCK分泌的动态过程

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