Method Article

Standardized Rat Coronary Ring Preparation and Real-Time Recording of Dynamic Tension Changes Along Vessel Diameter

DOI:

10.3791/64121

⸱

June 16th, 2022

 ,  ,  ,  ,  , 
1Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Basic Medicine, Chengdu University of Traditional Chinese Medicine, 3Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine

In This Article

Summary

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The present protocol describes the wire myograph technique for measuring vascular reactivity of the rat coronary artery.

Abstract

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As a key event of cardiovascular system diseases, coronary artery disease (CAD) has been widely regarded as the main culprit of atherosclerosis, myocardial infarction, and angina pectoris, which seriously threaten the life and health of people all over the world. However, how to record the dynamic biomechanical characteristics of isolated blood vessels has long puzzled people. Meanwhile, precise positioning and isolation of coronary arteries to measure in vitro dynamic vascular tension changes have become a trend in CAD drug development. The present protocol describes the macroscopic identification and microscopic separation of rat coronary arteries. The contraction and dilation function of the coronary artery ring along the vessel diameter was monitored using the established multi myograph system. The standardized and programmed protocols of coronary ring tension measurement, from sampling to data acquisition, tremendously improve the repeatability of the experimental data, which ensures the authenticity of vascular tension records after physiological, pathological, and drug intervention.

Introduction

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Coronary artery disease (CAD) has been widely recognized and concerned as a typical and representative cardiovascular disease, being the leading cause of death in both developed and developing countries1,2. As a blood and oxygen supply route for normal cardiac physiological function, circulating blood enters and nourishes the heart through two main coronary arteries and a blood vascular network on the surface of the myocardium3,4. Cholesterol and fat deposits in the coronary arteries cut off the heart's blood supply and the violent inflammatory res....

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Protocol

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The animal protocol was reviewed and approved by the Management Committee from Chengdu University of Traditional Chinese Medicine (Record No. 2021-11). Male Sprague Dawley (SD) rats (260-300 g, 8-10 weeks old) were used for the present study. The rats were kept in an animal chamber and were free to drink and eat during the experiment.

1. Solution preparation

  1. Prepare physiological salt solution (PSS) by dissolving 118 mM of NaCl, 4.7 mM of K+, 2.5 mM of CaCl2, 1.2 mM of KH2PO4, 1.2 mM of MgCl2∙6H2O, 25 mM of NaHCO3, 11 mM of D-gluc....

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Results

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Anatomically positioned, rat coronary arteries distributed and hidden deep in myocardial tissue were not easily recognized. By comparing the coronary arteries of humans (Figure 1A) and rats (Figure 1B), rapid and accurate separation of rat coronary arteries was conducted according to the sampling process in Figure 2. After precisely locating the right auricle, pulmonary artery, and apex from the front under an optical microscope, th.......

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Discussion

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The disturbance of coronary microcirculation, which involves a wide range of patients with CAD, has been gradually recognized and concerned the basis for adequate myocardium perfusion. Considering the serious complications of sudden coronary heart disease and cardiovascular disease, timely drug prevention and treatment are extremely important for a clinical individual with CAD17. Inevitably, the secrecy of coronary artery anatomy and the complexity of its physiological structure have severely rest.......

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Disclosures

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The authors have nothing to disclose.

Acknowledgements

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This work was supported by the Key R&D project of Sichuan Provincial Science and Technology Plan (2022YFS0438), the National Natural Science Foundation of China (82104533), the China Postdoctoral Science Foundation (2020M683273), and the Science & Technology Department of Sichuan Province (2021YJ0175).

....

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
ApigeninSangon Biotech Co., Ltd., Shanghai, China150731
CaCl2Sangon Biotech Co., Ltd., Shanghai, ChinaA501330
D-glucoseSangon Biotech Co., Ltd., Shanghai, ChinaA610219
HEPESXiya Reagent Co., Ltd., Shandong, ChinaS3872
KClSangon Biotech Co., Ltd., Shanghai, ChinaA100395
KH2PO4Sangon Biotech Co., Ltd., Shanghai, ChinaA100781
LabChart Professional version 8.3 ADInstruments, Australia—
MgCl2·6H2OSangon Biotech Co., Ltd., Shanghai, ChinaA100288
Multi myograph system Danish Myo Technology, Aarhus, Denmark620M
NaClSangon Biotech Co., Ltd., Shanghai, ChinaA100241
NaHCO3Sangon Biotech Co., Ltd., Shanghai, ChinaA100865
Steel wiresDanish Myo Technology, Aarhus, Denmark400447
U46619Sigma, USAD8174

References

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  1. Malakar, A. K., et al. A review on coronary artery disease, its risk factors, and therapeutics. Journal of Cellular Physiology. 234 (10), 16812-16823 (2019).
  2. Murray, C., et al.

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Tags

Standardized Rat Coronary RingDynamic Tension ChangesCardiovascular DiseasesVessel DiameterAnatomical LocationThoracic ArteryMesenteric ArteryOptical Anatomic MicroscopeLeft Coronary ArteryRight Coronary ArteryPhysiological Salt SolutionArterial Tension RecordingDissection TechniqueSurgical Scissors