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Using An In Vitro Tissue Perfusion System to Detect the Functional Activities of Isolated Intestinal Tubes in Real Time
In This Article
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 literature14,15,16,17. 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.......
Representative Results
The first part of the study focuses on the process of separating isolated intestinal tubes from the body and converting them into 2 cm tubes in vitro. This process is illustrated in detail in Figure 1. The second part involves the suspension and standardization of the isolated intestinal tube ring. The success of this process is demonstrated in Figure 2, which shows the automatic rhythmic contraction of a normal tube. Lastly, the study examines the effe.......
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 contraction20,21. 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.......
Acknowledgements
This work was supported by the Special Talent Program of Chengdu University of Traditional Chinese Medicine for "Xinglin Scholars and Discipline Talents Research Promotion Plan" (33002324).
....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 | — |
References
- Jiang, Z., et al. Therapeutic role of wuda granule in gastrointestinal motility disorder through promoting gastrointestinal motility and decreasing inflammatory level. Front Pharmacol. 14, 1-15 (2023).
- Talley, N. J.
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