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

doi:10.12300/j.issn.1674-5817.2022.024

Abstract

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

CLC Number:

Q95-33

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.

Figures/Tables 5

References 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