Using An In Vitro Tissue Perfusion System to Detect the Functional Activities of Isolated Intestinal Tubes in Real Time

Summary

We present a method for isolating rat intestinal tubes and assessing the impact of drugs on their tension, frequency, and amplitude in vitro. This method offers a valuable approach for researchers investigating intestinal tubes.

Abstract

Gastrointestinal diseases, which have a high incidence, pose considerable challenges for humans. The small intestine is integral to food and drug digestion and absorption and plays a crucial role in treating these diseases. The intestinal tube movement experiment, a common and essential in vitro method, is utilized to study gastrointestinal dynamics. This includes the preparation of the isolated intestinal tube, as well as the suspension of the prepared intestinal tube in the bath and its connection to a signal detector. This is followed by the recording and analysis of a series of parameters, such as tension, which can be used to assess intestinal motor function, as well as considerations for keeping the intestinal tube active in vitro. The standardized program from sampling to data collection greatly improves the repeatability of the experimental data and ensures the authenticity of the recording of intestinal tension after physiological, pathological, and drug intervention. Here we present the key problems in experimental operation and a valuable reference experimental protocol for studying drugs that regulate gastrointestinal motility.

Introduction

Gastrointestinal diseases, a prevalent condition, gravely impact human life and health¹. Gastrointestinal motility disorder is an important part of functional gastrointestinal diseases, manifesting primarily in debilitating symptoms, delayed gastric emptying, and severe gastric issues². It can disrupt gastrointestinal coordination, hinder gastric emptying, impact intestinal food intolerance, and even cause functional obstruction in the small or large intestine³. For patients undergoing gastrointestinal surgery, this disorder can directly lead to intestinal failure. Moreover, intestinal disorder is....

Protocol

This protocol is derived from previously published literature^14,15,16,17. Male Sprague Dawley (SD) rats (260-300 g, 8-10 weeks old) were used for the present study. The animal protocol was reviewed and approved by the Management Committee from Chengdu University of Traditional Chinese Medicine (Record No. 2023017). Prior to the experiment, the rats were instructed to fast for 24 h. During the e.......

Discussion

Gastrointestinal motility is accomplished by a series of precisely coordinated smooth muscle contractions and relaxations. This process involves rhythmic contraction of one group of muscle groups, coordinated contraction of multiple groups, and special propulsive contraction^20,21. The occurrence of gastrointestinal motility disorders may be associated with dysfunctions at different levels, such as the central nervous system, autonomic nervous system, enteric nerv.......

Materials

Name	Company	Catalog Number	Comments
Acetylcholine	Sigma, USA	A6625
atropine	Sangon Biotech Co., Ltd., Shanghai, China	IA06501
Barium chloride	Macklin Biochemical Co.,Ltd.,Shanghai, China	B861682
CaCl2	Sangon Biotech Co., Ltd., Shanghai, China	A501330
D-glucose	Sangon Biotech Co., Ltd., Shanghai, China	A610219
drawing software	GraphPad Software, San Diego, California, USA	—
Epinephrine	Sigma, USA	E4642
HEPES	Xiya Reagent Co., Ltd., Shandong, China	S3872
In vitro tissue perfusion system	PowerLab, ADInstruments, Australia	ML0146
KCl	Sangon Biotech Co., Ltd., Shanghai, China	A100395
KH2PO4	Sangon Biotech Co., Ltd., Shanghai, China	A100781
LabChart Professional version 8.3	ADInstruments, Australia	—
MgCl2·6H2O	Sangon Biotech Co., Ltd., Shanghai, China	A100288
NaCl	Sangon Biotech Co., Ltd., Shanghai, China	A100241
NaHCO3	Sangon Biotech Co., Ltd., Shanghai, China	A100865
nifedipine	Macklin Biochemical Co.,Ltd.,Shanghai, China	N5087
statistical analysis software	GraphPad Software, San Diego, California, USA	—
Surgical sutures	Johnson, USA	—