Long and new coronary myocardial injury real-time monitoring method based on biosensing system

Abstract

The invention relates to a long and new coronary myocardial injury real-time monitoring method based on a biosensing system. According to the method, the electrophysiological characteristics of myocardial cells infected by new coronavirus are monitored in real time mainly through a multi-electrode array module, and the dynamic change of calcium ions and the cell survival rate and apoptosis condition are detected in combination with an optical signal living cell imaging technology. By means of the method, researchers can simulate the infection process of the long new coronavirus on myocardial cells in an in-vitro system, and myocardial cell electrophysiological abnormality, calcium ion internal loss regulation and cell damage mechanisms caused by the virus can be deeply analyzed. The core technology of the invention comprises MEA-based electrophysiological signal acquisition and optical signal-based calcium ion flow monitoring. Research results show that the model method can effectively reveal the cytological mechanism of the new coronary myocardial injury, a powerful tool is provided for basic research of new coronary related cardiovascular diseases, and a solid foundation is laid for subsequent drug screening and development of treatment schemes.

Description

Technical Field

[0001] The present invention relates to the field of biomedical engineering, and in particular to a real-time monitoring method for long-term coronary myocardial injury based on a biosensor system. Background Art

[0002] Post-acute sequelae (PASC) of SARS-CoV-2 (COVID-19) infection manifest as symptoms that persist beyond the initial phase of COVID-19 infection, affecting an estimated 10-30% of survivors worldwide. These symptoms significantly impact cardiovascular health, leading to myocardial damage and arrhythmias, greatly reducing patients’ quality of life. Existing diagnostic techniques, such as echocardiography and cardiac magnetic resonance imaging, are unable to reveal the effects of the disease at a high-resolution cellular level, so the underlying mechanisms of PASC-induced cardiac dysfunction remain unclear, hindering the development of corresponding effective treatments. Therefore, the establishment of an efficient and practical in vitro platform for studying PASC-induced myocardial damage is critical to advance the development of precise diagnostic and therapeutic strategies.

[0003] Pseudoviruses are engineered viruses that mimic SARS-CoV-2 infection by expressing spike proteins but lack the ability to replicate and cause disease. Pseudoviruses help to safely explore the effects of multiple antibodies or sera on viral entry mechanisms, thereby accelerating the development and evaluation of COVID-19 vaccines. In addition, pseudoviruses are essential for elucidating the dynamic process of viral entry into host cells and studying the molecular interactions between viral spike proteins and host receptors such as ACE2. These insights are fundamental for developing precise interventions to slow the spread of infection. Pseudoviruses also play a vital role in the field of drug discovery. As an important tool for high-throughput screening of antiviral agents, their ability to promote the discovery of compounds with potential to inhibit viral entry, thereby accelerating the advancement of treatment options to clinical trials. With their nanoscale size, pseudoviruses provide a versatile platform at the intersection of nanotechnology and virology to advance the study of viral pathogenic mechanisms and prepare for future epidemics.

[0004] To elucidate the impact of COVID-19 on the cardiovascular system, current cutting-edge clinical and experimental methods play a key role. Clinically, myocardial function and damage in infected patients can be rigorously assessed through imaging and specific biomarker detection technologies. However, these traditional tools are insufficient to capture the persistent pathophysiological dysfunction that occurs during the infection process. To address these clinical questions, complex in vitro systems using primary cardiomyocytes and cardiomyocyte cell lines are essential for studying the direct cardiac effects of SARS-CoV-2 infection. These systems reveal the mechanisms of virus-induced inflammation and myocardial damage, but often fail to reproduce the complex specific responses and chronic manifestations of PASC. Long-term dynamic microelectrode array (MEA) recording of cardiomyocyte electrophysiological activity can make up for the data missing from traditional detection methods. By providing persistent, high-resolution insights into action potentials and arrhythmia patterns, MEA technology has significantly promoted a comprehensive understanding of the long-term adaptation of cardiomyocytes to viral effects and therapeutic interventions. This approach is essential for studying persistent cardiac pathology and designing targeted treatment options. Summary of the invention

[0005] The purpose of the present invention is to provide a real-time monitoring method for long-term new coronary myocardial injury based on a biosensor system to address the deficiencies of the in vitro monitoring methods for virus-induced myocardial injury in the prior art in terms of long-term dynamic monitoring and multi-dimensional data acquisition. This method can monitor the electrophysiological characteristics of myocardial cells and the dynamic changes of calcium ions over a long period of time, thereby achieving a comprehensive assessment of myocardial injury after long-term new coronary infection, and can be used for the effectiveness study of antiarrhythmic drugs.

[0006] The objective of the present invention is achieved through the following technical solutions:

[0007] A real-time monitoring method for myocardial injury caused by long-term coronary pneumonia based on a biosensor system is implemented by the following steps:

[0008] (1) In vitro culture of cardiomyocytes on multi-electrode array MEA: cardiomyocytes are seeded on the MEA chip and cultured until the cells adhere to the wall and begin to beat spontaneously;

[0009] (2) SARS-CoV-2 pseudovirus infection: Add a pseudovirus expressing the SARS-CoV-2 spike protein to the cardiomyocyte culture medium to simulate the viral infection process;

[0010] (3) Electrophysiological signal monitoring: The electrophysiological signals of cardiomyocytes are recorded in real time through the MEA module, including changes in action potential and electric field potential;

[0011] (4) Calcium ion flow monitoring: Using optical signal live cell imaging technology and calcium ion fluorescent probes, the dynamic changes of calcium ion concentration in cardiomyocytes can be monitored in real time;

[0012] (5) Dynamic monitoring: Dynamically monitor the changes in electrophysiological signals and calcium ion flow of cardiomyocytes after viral infection for N days, with N>5, to capture the dynamic process of chronic myocardial damage caused by long-term COVID-19 and evaluate cardiomyocyte activity.

[0013] Specifically, the cardiomyocytes in step (1) are primary cardiomyocytes or cardiomyocytes differentiated from induced pluripotent stem cells (iPSCs).

[0014] Specifically, the pseudovirus in step (2) includes a lentivirus expressing the SARS-CoV-2 spike protein and its variants, which is used to simulate the infection of the new coronavirus and can be used to screen and evaluate the cardiotoxicity of anti-new coronavirus drugs.

[0015] Specifically, the pseudovirus in step (2) can be adjusted to simulate different degrees of viral infection, and its concentration range is 10 4 Up to 10 9 IU / ml.

[0016] Specifically, the MEA module in step (3) comprises 8 to 5000 electrodes, the electrode diameter is 10 to 30 μm, and the distribution density is 100 to 1000 electrodes / cm 2 , used to monitor the electrical activity of several cardiomyocytes simultaneously.

[0017] Specifically, the optical signal live cell imaging technology uses a calcium ion fluorescent probe, which emits a fluorescent signal by binding to calcium ions, and then monitors the dynamic changes of calcium ion release; the dynamic changes of calcium ion influx are monitored using a high-sensitivity fluorescence microscope, and the imaging speed reaches more than 10 frames per second to capture the dynamic changes of calcium ions in cells; the calcium ion fluorescent probes include but are not limited to the following four: Fura-2 AM, Fluo-4 AM, Rhod-2 AM and Calcium Green-1 AM.

[0018] Furthermore, the cardiomyocyte activity assessment includes labeling apoptotic cells and viable cells with Annexin-V and DAPI fluorescent dyes, and analyzing them using a flow cytometer.

[0019] Specifically, the pseudovirus 0-5×10 7 The dose-response relationship of virus damage to cardiomyocytes was evaluated within the IU dose range.

[0020] Specifically, the monitoring process in step (5) is carried out under standard culture conditions, maintaining a temperature of 37° C. and a CO 2 concentration of 5%.

[0021] Furthermore, on the basis of virus infection of cardiomyocytes, the effect of antiarrhythmic drugs on electrophysiological abnormalities of cardiomyocytes can be used to evaluate their cardioprotective effect, and the changes in the electrical activity and calcium ion dynamics of cardiomyocytes before and after drug treatment can be analyzed by MEA and calcium ion flow monitoring to quantify the protective effect of antiarrhythmic drugs; the antiarrhythmic drugs include but are not limited to the following four: Nifedipine, Amiodarone, Propranolol and Lidocaine.

[0022] The beneficial effects of the present invention are as follows: the method of the present invention is easy to operate and can be used for long-term dynamic monitoring of physiological changes of myocardial cells under SARS-CoV-2 pseudovirus infection, which is conducive to a comprehensive assessment of the impact of the virus on myocardial cells. The present invention provides a real-time monitoring model method of long-term new coronary myocardial injury based on a multimodal biosensor system, which can obtain high-quality electrophysiological signals of myocardial cells and dynamic changes of calcium ions, respectively, and can extend the monitoring time, thereby achieving accurate and long-term monitoring of myocardial cell function, and providing an important experimental platform and technical support for the research and drug development of long-term new coronary heart diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The present invention is further described below in conjunction with the accompanying drawings and embodiments;

[0024] Figure 1 This is a diagram of in vitro culture of myocardial cells on a biosensor system of the present invention;

[0025] Figure 2 This is a diagram of the SARS-CoV-2 pseudovirus infecting primary cardiomyocytes of the present invention;

[0026] Figure 3 This is a graph of electrical signals and calcium ion flow detection of primary myocardial cells infected with a pseudo-SARS-CoV-2 long-term disease of the present invention;

[0027] Figure 4 This is an electrical signal and calcium ion flow detection diagram of nifedipine against SARS-CoV-2 pseudo-disease infection of the present invention. DETAILED DESCRIPTION

[0028] The present invention provides a real-time monitoring model method of long-term new coronary myocardial injury based on a multimodal biosensor system, including: inoculating cardiomyocytes on a multi-electrode array (MEA) chip for cultivation; using a pseudovirus expressing the SARS-CoV-2 spike protein to infect cardiomyocytes; monitoring the electrophysiological signals of cardiomyocytes and the dynamic changes of calcium ion flow respectively; through the application of antiarrhythmic drugs, evaluating their protective effects on abnormal cardiomyocyte function (cardiomyocyte activity evaluation includes labeling apoptotic cells and surviving cells with Annexin-V and DAPI fluorescent dyes, and analyzing them using a flow cytometer; obtaining cell apoptosis rate and survival rate). The model method is simple to operate, can be used for long-term dynamic monitoring of physiological changes of cardiomyocytes under viral infection, and can systematically evaluate the effectiveness of antiarrhythmic drugs. The present invention is further described below in conjunction with the accompanying drawings and specific embodiments.

[0029] A real-time monitoring model method of long-term myocardial injury caused by COVID-19 based on a multimodal biosensor system, comprising the following steps:

[0030] (1) In vitro culture of cardiomyocytes on MEA:

[0031] The cardiomyocytes were seeded on the MEA chip and cultured until the cells adhered to the wall and began to beat spontaneously, ensuring that the cells were in a stable state.

[0032] Furthermore, the cardiomyocytes may be primary cardiomyocytes or cardiomyocytes differentiated from induced pluripotent stem cells (iPSCs).

[0033] (2) SARS-CoV-2 pseudovirus infection:

[0034] Pseudovirus expressing SARS-CoV-2 spike protein was added to the culture medium of cardiomyocytes to simulate the viral infection process.

[0035] Furthermore, the concentration of the pseudovirus can be adjusted to simulate different degrees of viral infection, and the commonly used concentration range is 10 4 Up to 10 9 IU / ml.

[0036] (3) Electrophysiological signal monitoring:

[0037] The electrophysiological signals of cardiomyocytes, including changes in action potential and electric field potential, are recorded in real time through the MEA module.

[0038] Furthermore, the MEA module comprises 8 to 5000 electrodes, the electrode diameter is 10 to 30 μm, and the distribution density is 100 to 1000 / cm 2 , used to monitor the electrical activity of multiple cardiomyocytes simultaneously.

[0039] (4) Calcium ion flow monitoring:

[0040] Using optical signal live cell imaging technology and calcium ion fluorescent probes, the dynamic changes of calcium ion concentration in cardiomyocytes can be monitored in real time.

[0041] The calcium ion fluorescent dyes used include but are not limited to:

[0042] Fura-2 AM: A widely used ratiometric calcium dye that can accurately measure intracellular calcium concentrations through UV excitation.

[0043] Fluo-4 AM: A highly sensitive calcium dye that is excited by visible light and is suitable for high-throughput screening and real-time calcium imaging.

[0044] Rhod-2 AM: A red fluorescent calcium ion dye with high photostability, suitable for detecting changes in calcium ions in mitochondria.

[0045] Calcium Green-1 AM: A highly sensitive and photostable calcium dye suitable for long-term calcium imaging experiments.

[0046] Furthermore, calcium ion flow was monitored using a high-sensitivity fluorescence microscope with an imaging speed of more than 10 frames per second to capture the rapid dynamic changes of calcium ions within the cell.

[0047] (5) Long-term dynamic monitoring:

[0048] This method captures the dynamic process of chronic myocardial damage caused by long-term (5 days or more) dynamic monitoring of changes in electrophysiological signals and calcium ion flow in cardiomyocytes after viral infection.

[0049] Furthermore, the monitoring process can be carried out under standard culture conditions, maintaining a temperature of 37° C. and a CO 2 concentration of 5%.

[0050] Antiarrhythmic Drug Assessment:

[0051] Based on the virus infection of cardiomyocytes, the effects of commonly used antiarrhythmic drugs on electrophysiological abnormalities of cardiomyocytes were studied to evaluate their cardioprotective effects. The antiarrhythmic drugs studied include but are not limited to:

[0052] Nifedipine: A calcium channel blocker widely used in the treatment of hypertension and angina pectoris, which alleviates electrophysiological abnormalities of myocardial cells by inhibiting calcium ion influx.

[0053] Amiodarone: A broad-spectrum antiarrhythmic drug that prevents and treats arrhythmias by prolonging the duration of the action potential and reducing myocardial excitability.

[0054] Propranolol: A non-selective beta-blocker that prevents arrhythmias by lowering heart rate and reducing myocardial oxygen consumption.

[0055] Lidocaine: A sodium channel blocker that inhibits the influx of sodium ions and reduces the conduction of abnormal electrical signals. It is often used to treat acute arrhythmias.

[0056] Furthermore, MEA and calcium ion flow monitoring were used to analyze the changes in myocardial cell electrical activity and calcium ion dynamics before and after drug treatment to quantify the protective effect of the drug.

[0057] Embodiment 1:

[0058] A real-time monitoring model method of long-term myocardial injury caused by COVID-19 based on a multimodal biosensor system, comprising the following steps:

[0059] (1) In vitro culture of cardiomyocytes on MEA:

[0060] like Figure 1 As shown in Figure 1, MEA devices are mainly manufactured by photolithography. The substrate of the device is made of four-inch quartz glass. First, a 2.5μm thick positive photoresist is sprayed on the glass substrate at a speed of 3000rpm. After soft baking at 120℃ for two minutes, it is developed with RZX3038 developer for 35 seconds and then passed through a mask aligner at 300mJ / cm 2 The conductive design pattern was formed by exposing with an intensity of 100 nm to a light source. A layer of 10 nm titanium / 100 nm gold was deposited on the substrate using magnetron sputtering technology, and then the photoresist was stripped with acetone. Next, a 5 μm thick layer of SU-8 2005 photoresist was spin-coated at a speed of 3000 rpm and soft-baked at 95 °C for two minutes. A mask aligner was then used to apply 160 mJ / cm 2 The SU-8 layer was exposed to an intensity of 1000 nm and post-exposure baked at 95°C for three minutes. After one minute of development, the layer was rinsed with propylene glycol methyl ether acetate (PGMEA), cleaned with isopropyl alcohol, dried with a nitrogen spray gun, and baked at 150°C for 30 minutes. Finally, the substrate was cut into 25 pieces of 2×2 cm 2sheets, each containing 32 microelectrodes. The microelectrode array chip was then attached to a custom-designed printed circuit board (PCB) adapter using polydimethylsiloxane (PDMS). Conductive silver glue was used to electrically connect the electrode pads on the chip to the corresponding pads on the PCB. A glass ring, which served as a cell culture pool, was fixed to the chip using PDMS. The PCB was equipped with pin connectors to interface with the self-developed biosensing system. In a biological safety cabinet, the device was sterilized by ultraviolet irradiation for 2 hours after rinsing in 75% ethanol. Before cell inoculation, a 10μg / mL fibronectin solution was coated on the device and incubated overnight in a 4°C refrigerator.

[0061] Primary neonatal cardiomyocytes were obtained from 1-day-old Wistar rats. After disinfection with 75% ethanol, the heart tissue was quickly excised and placed in Hanks balanced salt solution (HBSS), and then rinsed with ice-cold Dulbecco's modified Eagle's medium (DMEM). The ventricular tissue was then cut into approximately 1 mm 3 Small pieces were placed in HBSS containing 0.05% type II collagenase and 0.07% trypsin for digestion. The digestion process was repeated ten to twelve times in a 37°C, 5% CO2 incubator. After each digestion, the supernatant was collected and centrifuged at 1000rpm for 5 minutes. The cardiomyocytes were purified by differential adhesion method, and the purification process was repeated twice to increase the purity. The isolated cardiomyocytes were then resuspended in DMEM supplemented with 10% fetal bovine serum, 100U / mL penicillin and 100μg / mL streptomycin and subsequently inoculated onto the MEA device. To maintain optimal growth conditions, the culture medium was replaced every 48 hours.

[0062] (2) SARS-CoV-2 pseudovirus infection:

[0063] Will Figure 1 Primary neonatal rat cardiomyocytes were cultured and SARS-CoV-2 pseudovirus expressing GFP (7.26×10 8 IU / ml, 5μL / well, 10μL / well and 20μL / well) were infected at the optimized MOI (multiplicity of infection). To promote viral entry into cells, 10μg / mL polybromide was added. Subsequently, the cells were incubated at 37°C and 5% CO2 for 48 hours, and GFP expression in infected cardiomyocytes was observed by fluorescence microscopy and image capture was performed. Figure 2 This is the GFP expression detection after SARS-CoV-2 pseudovirus infected primary cardiomyocytes. The figure shows that the pseudovirus can enter primary cardiomyocytes, and GFP is expressed in a dose-dependent manner 48 hours after infection.

[0064] (1) Long-term dynamic monitoring of electrical signals and calcium ion currents

[0065] The extracellular action potential of myocardial cells Figure 1 The custom-made biosensing system was used for recording, which integrates electroporation and signal recording functions. Figure 3 As shown. First, the primary amplifier module captures and amplifies the weak electrical signals in the device tenfold, and then transmits them through a signal conditioning circuit designed for precise electrophysiological recording. These signals are then further enhanced fifty times by the secondary amplifier and sampled at a frequency of 20kHz by the analog-to-digital converter (ADC) within the high-speed data acquisition (DAQ) card. To reduce noise, a bandpass filter of 1Hz to 7.5kHz is used. The detection range of the DAQ card is -5 to +5V, and the ADC provides 16-bit resolution with a sensitivity of 153μV. The operation of the entire system, including signal recording via USB communication, and subsequent data display, acquisition, storage, and processing, is controlled by a custom-developed LabVIEW program. All cell experiments were performed in a controlled environment at 37°C and 5% CO2 to ensure conditions suitable for long-term monitoring. Two days after the cells were inoculated into the device, they were infected with SARS-CoV-2 pseudovirus. Extracellular electrical signals were initially recorded without pseudovirus infection to establish a baseline, and then the signals were continuously recorded for five days during infection.

[0066] Calcium ion flow detection: The treated cardiomyocytes were cultured in 24-well plates. Rhod-4 AM dye was dissolved in DMSO to prepare a 5mM stock solution. The cells were washed with HBSS and incubated in HBSS containing Rhod-4 AM working solution (2μM) for 30 minutes at 37°C, protected from light. After incubation, the cells were washed with HBSS and allowed to stand for 30 minutes to stabilize. Fluorescence imaging was then performed using a fluorescence microscope. Changes in fluorescence intensity were recorded to monitor calcium ion flow in real time.

[0067] pass Figure 3 The electrical and optical signal monitoring shown in the figure establishes a long-term dynamic monitoring model of long-term coronary myocardial injury, laying the foundation for the systematic evaluation of the effectiveness of antiarrhythmic drugs.

[0068] Embodiment 2:

[0069] Antiarrhythmic drug evaluation. Use Figure 1The primary neonatal rat cardiomyocytes were first cultured on the MEA chip and stabilized to spontaneous beating, and then a 10 μL dose of SARS-CoV-2 pseudovirus was added to infect the cardiomyocytes to simulate chronic myocardial damage associated with long COVID-19. At the same time as the virus infection, 100 nM of nifedipine was added to the experimental group, and the electrophysiological activity and calcium ion flow of the cardiomyocytes were monitored in real time over the next five days. The MEA module records the action potential amplitude and discharge frequency of the cardiomyocytes, and uses Rhod-4 AM dye to label the calcium ions in the cardiomyocytes, and the dynamic changes of calcium ions are observed by fluorescence microscopy. In addition, morphological evaluation was performed to analyze the effect of nifedipine on the morphology and viability of cardiomyocytes. Based on the results of electrophysiological signals, calcium ion dynamics and morphological evaluation, nifedipine showed significant cardioprotective effects in inhibiting arrhythmias caused by SARS-CoV-2 pseudovirus and restoring calcium ion homeostasis, especially in the early and late stages after infection, showing strong therapeutic potential.

[0070] Figure 4 The electrical signal diagram and calcium ion flow change curve of nifedipine in suppressing arrhythmia monitored in this example. Combined with morphological analysis, the effect of nifedipine on myocardial injury caused by long-term coronary pneumonia was comprehensively evaluated.

[0071] In summary, the above embodiments are only for illustrating the principles and effects of the present invention and do not constitute a limitation on the implementation methods. For those skilled in the art, various modifications and improvements can be made without departing from the spirit and scope of the present invention, and these modifications and improvements should be regarded as contents within the protection scope of the present invention.

Claims

1. A real-time monitoring method for myocardial injury caused by coronary pneumonia based on a biosensor system, characterized in that: The method is implemented by the following steps: (1) In vitro culture of cardiomyocytes on multi-electrode array MEA: cardiomyocytes are seeded on the MEA chip and cultured until the cells adhere to the wall and begin to beat spontaneously; (2) SARS-CoV-2 pseudovirus infection: Add a pseudovirus expressing the SARS-CoV-2 spike protein to the cardiomyocyte culture medium to simulate the viral infection process; (3) Electrophysiological signal monitoring: The electrophysiological signals of cardiomyocytes are recorded in real time through the MEA module, including changes in action potential and electric field potential; (4) Calcium ion flow monitoring: Using optical signal live cell imaging technology and calcium ion fluorescent probes, the dynamic changes of calcium ion concentration in cardiomyocytes can be monitored in real time; (5) Dynamic monitoring: Dynamically monitor the changes in electrophysiological signals and calcium ion flow of cardiomyocytes after viral infection for N days, with N>5, to capture the dynamic process of chronic myocardial damage caused by long-term COVID-19 and evaluate cardiomyocyte activity.

2. The method according to claim 1, characterized in that The cardiomyocytes in step (1) are primary cardiomyocytes or cardiomyocytes differentiated from induced pluripotent stem cells (iPSCs).

3. The method according to claim 1, characterized in that The pseudovirus in step (2) includes a lentivirus expressing the SARS-CoV-2 spike protein and its variants, which is used to simulate the infection of the new coronavirus and can be used to screen and evaluate the cardiotoxicity of anti-new coronavirus drugs.

4. The method according to claim 1, characterized in that: In step (2), the pseudovirus can be adjusted to simulate different degrees of viral infection, and its concentration range is 10 4 Up to 10 9 IU / ml.

5. The method according to claim 1, characterized in that The MEA module in step (3) comprises 8 to 5000 electrodes, the electrode diameter is 10 to 30 μm, and the distribution density is 100 to 1000 electrodes / cm 2 , used to monitor the electrical activity of several cardiomyocytes simultaneously.

6. The method according to claim 1, characterized in that The optical signal live cell imaging technology uses a calcium ion fluorescent probe to emit a fluorescent signal by binding to calcium ions, thereby monitoring the dynamic changes of calcium ion release; the dynamic changes of calcium ion influx are monitored using a high-sensitivity fluorescence microscope, with an imaging speed of more than 10 frames per second to capture the dynamic changes of calcium ions in cells; the calcium ion fluorescent probes include but are not limited to the following four: Fura-2AM, Fluo-4 AM, Rhod-2 AM and Calcium Green-1 AM.

7. The method according to claim 1, characterized in that The cardiomyocyte activity assessment includes labeling apoptotic cells and viable cells with Annexin-V and DAPI fluorescent dyes, and analyzing them using flow cytometry.

8. The method according to claim 1, characterized in that The pseudovirus 0-5×10 7 The dose-response relationship of virus damage to cardiomyocytes was evaluated within the IU dose range.

9. The method according to claim 1, characterized in that: The monitoring process in step (5) is carried out under standard culture conditions, maintaining a temperature of 37° C. and a CO 2 concentration of 5%.

10. The method according to claim 1, characterized in that On the basis of virus infection of cardiomyocytes, the effect of antiarrhythmic drugs on electrophysiological abnormalities of cardiomyocytes can be used to evaluate their cardioprotective effect, and the changes in the electrical activity and calcium ion dynamics of cardiomyocytes before and after drug treatment can be analyzed by MEA and calcium ion flow monitoring to quantify the protective effect of antiarrhythmic drugs; the antiarrhythmic drugs include but are not limited to the following four: Nifedipine, Amiodarone, Propranolol and Lidocaine.

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