大鼠肠系膜淋巴液持续引流新方法的初步建立

doi:10.12300/j.issn.1674-5817.2022.024

实验动物与比较医学 ›› 2022, Vol. 42 ›› Issue (4): 267-274.DOI: 10.12300/j.issn.1674-5817.2022.024

• 动物实验技术与方法专题 • 下一篇

大鼠肠系膜淋巴液持续引流新方法的初步建立

张笑瑞¹(), 曹静², 吴倩倩¹, 康康¹, 陈国元¹, 吴宝金¹()()

^1.中国科学院分子细胞科学卓越创新中心动物实验技术平台, 上海 200031
^2.上海中医药大学科技实验中心实验动物中心, 上海 200031

收稿日期:2022-03-04 修回日期:2022-05-17 出版日期:2022-08-25 发布日期:2022-09-01
通讯作者: 吴宝金（1969—），男，博士，研究员，研究方向：小鼠遗传学和人类疾病动物模型。E-mail：baojin.wu@sibcb.ac.cn。ORCID：0000-0002-9768-8862
作者简介:张笑瑞（1987—），男，工程师，研究方向：人类疾病动物模型与新药临床前评价。E-mail：xiaorui.zhang@sibcb.ac.cn
基金资助:
中国科学院战略生物资源计划实验动物平台项目-2022年度实验动物模型研发子课题(KFJ-BRP-005)

A Preliminary Method for Continuous Drainage of Mesenteric Lymph Fluid in Rats

Xiaorui ZHANG¹(), Jing CAO², Qianqian WU¹, Kang KANG¹, Guoyuan CHEN¹, Baojin WU¹()()

^1.Animal Core Facility, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences; University of Chinese Academy of Sciences, Shanghai 200031, China
^2.Experiment Animal Center, Experiment Center for Science and Technology, Shanghai University of Traditional Chinese Medicine, Shanghai 200031, China

Received:2022-03-04 Revised:2022-05-17 Published:2022-08-25 Online:2022-09-01
Contact: WU Baojin (ORCID: 0000-0002-9768-8862), E-mail: baojin.wu@sibcb.ac.cn

摘要/Abstract

摘要：

目的通过建立大鼠肠系膜淋巴管-颈静脉辅助回流方法，实现肠系膜淋巴液的持续引流。方法将16只雄性SD大鼠随机均分为2组。对照组大鼠进行十二指肠和肠系膜淋巴管双插管手术，术后收集肠系膜淋巴液。实验组大鼠进行颈静脉和肠系膜淋巴管双插管手术，建立肠系膜淋巴管-颈静脉循环模型。术后第7天利用清醒活动装置收集肠系膜淋巴液，记录各组大鼠的肠系膜淋巴液流量，检测其细胞成分及部分生化指标。结果成功建立了大鼠肠系膜淋巴管-颈静脉辅助回流方法，模型维持时间达7 d以上。实验组大鼠的肠系膜淋巴液流量多于对照组流量［（2.01±0.12）mL/h vs（0.92±0.09）mL/h，P＜0.01］。实验组淋巴液中淋巴细胞数和淋巴细胞百分率高于对照组（P＜0.01，P＜0.05），而中性粒细胞比例和单核细胞比例均低于对照组（P＜0.01）。实验组淋巴液中K⁺、Na⁺、CO₂和尿素水平均高于对照组（P＜0.01），而三酰甘油和P³⁺水平均低于对照组（P＜0.01）。结论肠系膜淋巴管-颈静脉辅助回流技术能够实现大鼠在清醒、不限制饮食和正常活动状态下，实时并长期收集淋巴液，避免了手术应激、全身麻醉或动物受限对实验结果的影响。

关键词: 肠系膜淋巴液, 插管, 辅助回流, SD大鼠

Abstract:

Objective To establish a novel sustained collection method for mesenteric lymph fluid by means of assisted reflux from the mesenteric-jugular lymphatic duct in rats. Methods Sixteen male Sprague-Dawley (SD) rats were randomly divided into two groups. The control group underwent duodenal and intestinal lymphatic duct cannulation, after which intestinal lymph fluid was collected. The experimental group underwent jugular vein and intestinal lymphatic duct cannulation to establish intestinal-jugular lymphatic duct assisted reflux. The intestinal lymph fluid was collected on the 7th day after the operation using an awake mobility device. The flow rate of intestinal lymph fluid was recorded, and its cellular components and some biochemical indicators were detected. Results Mesenteric-jugular lymphatic duct vein cannulation assisted reflux was successfully established in rats, and the rat models could be maintained for more than seven days. The intestinal lymph flow rate was （2.01±0.12） mL/h in the experimental group, which was higher than that of the control group [(0.92±0.09) mL/h, P＜0.01]. The number of lymphocytes (LYM#) and percentage of lymphocytes (LYM/%) in the experimental group were higher than those in the control group (P＜0.01). The percentage of neutrophils (NEUT/%) and percentage of monocytes (MONO/%) were lower than those in the control group (P＜0.01, P＜0.05, respectively). The concentrations of K⁺, Na⁺, CO₂, and urea in the lymph fluid of the experimental group were higher than those of the control group (P＜0.01). However, the concentrations of triacylglycerol (TG) and P³⁺ were lower than those of the control group (P＜0.01). Conclusion This novel method can achieve real-time and long-term collection of mesenteric lymph fluid in rats under the condition of being awake, unrestricted in diet and in normal state, avoiding the influence of surgical stress, general anesthesia or animal restraint on the experimental results.

Key words: Mesenteric lymph fluid, Cannulation, Assisted reflux, Sprague-Dawley rats

中图分类号:

Q95-33

张笑瑞, 曹静, 吴倩倩, 康康, 陈国元, 吴宝金. 大鼠肠系膜淋巴液持续引流新方法的初步建立[J]. 实验动物与比较医学, 2022, 42(4): 267-274.

Xiaorui ZHANG, Jing CAO, Qianqian WU, Kang KANG, Guoyuan CHEN, Baojin WU. A Preliminary Method for Continuous Drainage of Mesenteric Lymph Fluid in Rats[J]. Laboratory Animal and Comparative Medicine, 2022, 42(4): 267-274.

图/表 5

参考文献 22

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血细胞分类 Blood cell classification	对照组 Control group (n=8)	实验组 Experimental group (n=8)
WBC/(10⁹·L^-1)	7.48±3.45	14.01±9.93
NEUT#/(10⁹·L^-1)	0.79±0.42	0.45±0.20
LYM#/(10⁹·L^-1)	2.14±1.03	7.34±4.89^*
MONO#/(10⁹·L^-1)	4.54±2.36	6.21±5.09
EOS#/(10⁹·L^-1)	0.01±0.01	0.01±0.01
BASO#/(10⁹·L^-1)	0.01±0.01	0.01±0.00
NEUT/%	11.20±4.51	4.35±2.16^**
LYM/%	28.23±6.78	51.99±7.40^**
MONO/%	60.36±9.81	43.44±6.61^**
EOS/%	0.15±0.12	0.14±0.15
BASO/%	0.06±0.07	0.09±0.14

血细胞分类 Blood cell classification	对照组 Control group (n=8)	实验组 Experimental group (n=8)
WBC/(10⁹·L^-1)	7.48±3.45	14.01±9.93
NEUT#/(10⁹·L^-1)	0.79±0.42	0.45±0.20
LYM#/(10⁹·L^-1)	2.14±1.03	7.34±4.89^*
MONO#/(10⁹·L^-1)	4.54±2.36	6.21±5.09
EOS#/(10⁹·L^-1)	0.01±0.01	0.01±0.01
BASO#/(10⁹·L^-1)	0.01±0.01	0.01±0.00
NEUT/%	11.20±4.51	4.35±2.16^**
LYM/%	28.23±6.78	51.99±7.40^**
MONO/%	60.36±9.81	43.44±6.61^**
EOS/%	0.15±0.12	0.14±0.15
BASO/%	0.06±0.07	0.09±0.14

生化指标 Biochemical indicators	对照组 Control group (n=8)	实验组 Experimental group (n=8)
Cl^-c/(mmol·L^-1)	99.75±1.85	100.75±1.71
K⁺c/(mmol·L^-1)	4.75±0.74	5.74±0.21^**
Na⁺c/(mmol·L^-1)	136.75±1.48	139.63±1.58^**
P³⁺c/(mmol·L^-1)	3.79±0.18	3.49±0.11^**
Ca²⁺c/(mmol·L^-1)	2.65±0.11	2.66±0.05
Mg²⁺c/(mmol·L^-1)	0.90±0.13	0.93±0.07
UREA c/(mmol·L^-1)	5.13±0.33	5.75±0.66^*
CRE c/(µmol·L^-1)	23.56±8.91	24.61±3.50
URIC c/(mmol·L^-1)	37.81±21.46	51.19±8.85
GLU c/(mmol·L^-1)	12.83±3.01	14.66±1.55
TG c/(mmol·L^-1)	3.63±1.02	1.57±0.47^**
CHO c/(mmol·L^-1)	1.69±0.16	1.62±0.20
HDL c/(mmol·L^-1)	1.16±0.13	1.15±0.15
TP ρ/(g·L^-1)	47.05±2.35	45.31±1.19
ALB ρ/(g·L^-1)	24.81±1.91	23.51±0.66
CO₂c/(mmol·L^-1)	22.50±2.45	27.25±1.09^**

生化指标 Biochemical indicators	对照组 Control group (n=8)	实验组 Experimental group (n=8)
Cl^-c/(mmol·L^-1)	99.75±1.85	100.75±1.71
K⁺c/(mmol·L^-1)	4.75±0.74	5.74±0.21^**
Na⁺c/(mmol·L^-1)	136.75±1.48	139.63±1.58^**
P³⁺c/(mmol·L^-1)	3.79±0.18	3.49±0.11^**
Ca²⁺c/(mmol·L^-1)	2.65±0.11	2.66±0.05
Mg²⁺c/(mmol·L^-1)	0.90±0.13	0.93±0.07
UREA c/(mmol·L^-1)	5.13±0.33	5.75±0.66^*
CRE c/(µmol·L^-1)	23.56±8.91	24.61±3.50
URIC c/(mmol·L^-1)	37.81±21.46	51.19±8.85
GLU c/(mmol·L^-1)	12.83±3.01	14.66±1.55
TG c/(mmol·L^-1)	3.63±1.02	1.57±0.47^**
CHO c/(mmol·L^-1)	1.69±0.16	1.62±0.20
HDL c/(mmol·L^-1)	1.16±0.13	1.15±0.15
TP ρ/(g·L^-1)	47.05±2.35	45.31±1.19
ALB ρ/(g·L^-1)	24.81±1.91	23.51±0.66
CO₂c/(mmol·L^-1)	22.50±2.45	27.25±1.09^**

指标 Indicator	传统技术 Traditional technology	新技术 Novel technology
动物状态 State of the animal	麻醉或束缚状态	清醒、自由活动
淋巴液收集频率 Frequency of lymph collection	仅术后单次收集	实时、连续多次收集
给药方式 Route of administration	受限制，部分给药需通过手术，如胃插管	不限制，结合自动化仪器，可实现自动化给药和体液采集
动物饮食 Animal feeding state	禁食	自由饮食
原肠淋巴管循环通路 Circulation pathway of mesenteric lymphatic duct	阻断	建立新的肠系膜淋巴管-左颈浅静脉管循环通路
淋巴液流速 Flow rate of lymph fluid	不稳定，不能反映真实淋巴液流速	稳定，真实反映淋巴液流速
补液方式 Rehydration method	通过十二指肠插管以恒定速率补充	通过颈静脉插管补充，并根据淋巴液流速实时调整，维持机体体液平衡
肠系膜淋巴液收集 Collection of intestinal lymphatic fluid	部分	全部
造模成功率 Modeling success rate	较低	高

大鼠肠系膜淋巴液持续引流新方法的初步建立

A Preliminary Method for Continuous Drainage of Mesenteric Lymph Fluid in Rats

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