KR100991548B1 - system for measuring neurological changes in the ischemia of rat model - Google Patents

Abstract

The present invention relates to a real-time nervous system measurement system according to the Iskemia of the animal model, in particular, meets the optimal conditions of the animal (temperature and temperature of the brain) in the Ischemia state of the animal, and the induction of the Ischemia is made normally The present invention relates to a real-time nervous system measurement system according to Ischemia of an animal model that measures the amount of change in neurotransmitters in the brain while checking whether it is lost.

The constituent means constituting the real-time nervous system measurement system according to the present invention iskemia of the animal model, the estimia induced confirmation device for measuring the biological parameters of the animal in order to confirm the induction of ischemia of the animal, the es Ischemia maintenance optimal device for measuring and maintaining the biometric parameters of the animal in order to maintain the survival of the animal and optimize the real-time measurement of nerve changes in accordance with Chemia, neurotransmitters released in the ischemia state of the animal Neurotransmitter measuring device for measuring the amount of change, characterized in that it comprises a data analysis device for receiving and storing data from the devices.

Animal Models, Ischemia, Neurotransmitters, Glutamate

Description

System for measuring neurological changes in the ischemia of rat model

The present invention relates to a real-time nervous system measurement system according to the Iskemia of the animal model, in particular, meets the optimal conditions of the animal (temperature and temperature of the brain) in the Ischemia state of the animal, and the induction of the Ischemia is made normally The present invention relates to a real-time nervous system measurement system according to Ischemia of an animal model that measures the amount of change in neurotransmitters in the brain while checking whether it is lost.

Currently, various medical experiments are being conducted using animals as models, and in order to determine and use the data of the experiment using the animal model as valid data for medical use, the condition of the animals must be maintained during the experiment. That is, there is a need to maintain a constant normal temperature as well as to maintain a survival state.

In addition, in order to measure the amount of real-time change of the release of neurotransmitters in the escapia state, it is necessary to measure in real time whether the animal's escapia state is normally reached.

On the other hand, it is also necessary to measure how the changes in the neurotransmitters with excitatory toxicities released from the brain are changed among the neurological changes that occur when esketemia is induced.

In summary, in order to measure the glutamate level in the interstitial space of the animal in real time, it is possible to implement forebrain ischemia as completely as possible, and the animal is in a healthy state for several minutes. In addition to ischemia, it is also necessary to verify that it can withstand reperfusion.

As described above, in order to measure the neural change in the escapia state in real time with respect to the animal model, it is necessary to measure and maintain a comprehensive biometric parameter, and in this state, the amount of change of the neurotransmitter must be measured. The amount of change in glutamate, a neurotransmitter, cannot be measured under normal measurement and maintenance conditions.

The present invention was devised to solve the problems of the prior art as described above, and meets the optimal conditions of the animal (temperature and temperature of the brain) in the escapia state of the animal, and whether the induction of escapia is normally performed. The purpose of the present invention is to provide a real-time nervous system measurement system according to the escapia of animal models that measure the amount of change of neurotransmitters in the brain while checking.

The constituent means constituting the real-time nervous system measurement system according to the present invention iskemia in order to solve the above problems, measuring the biological parameters of the animal in order to confirm the induction of the animal (ischemia) Ischemia-induced confirmation device, Ischemia maintenance optimization device for measuring and maintaining the biological parameters of the animal in order to maintain the survival of the animal and optimize the real-time measurement of nerve changes in accordance with the Neurotransmitter measuring device for measuring the amount of change in neurotransmitter released in the escape state, characterized in that it comprises a data analysis device for receiving and storing data from the devices.

In addition, the apparatus for confirming the induction of Eschiaia includes an electroencephalogram measuring means for measuring the change in the brain wave of the animal, a brain blood flow measuring means for measuring the blood flow change in the animal brain, and a blood pressure measuring means for measuring the blood pressure change in the animal. Characterized in that made.

In addition, the measured electroencephalogram, brain blood flow and blood pressure values are converted into digital values through an A / D converter, and then stored in the data analysis device.

In addition, the Ischemia maintenance optimum device is to maximize the body temperature measurement and maintenance means and stroke volume to maintain the body temperature of the animal at a predetermined temperature in order to prevent the rapid temperature decrease of the animal in the escape state Characterized in that it comprises a means for measuring and maintaining the temperature of the brain to maintain the temperature of the brain at a predetermined temperature.

In addition, the neurotransmitter measuring device is characterized in that for measuring the amount of release of glutamate (glutamate) using a microdialysis (microdialysis) biosensor or CNT probe.

According to the real-time nervous system measurement system according to the present invention iskemia of the animal model having the above object and the solution means, meets the optimal condition conditions (temperature and temperature of the brain) of the animal in the escape state of the animal, Since the change in the amount of neurotransmitters in the brain is measured while checking whether the chemia induction has been performed normally, there is an advantage in that it is possible to measure the change in the amount of changes in the neurotransmitter glutamate in real time in an optimal state.

Hereinafter, with reference to the accompanying drawings will be described in detail a preferred embodiment of the real-time nervous system measurement system according to the Esmemia of the animal model of the present invention having the above problems, solving means and effects.

The present invention is based on the 11VO model as an animal model that allows minimal modifications in physiological conditions of animals while maintaining hemodynamically stable breathing and self-sustaining life, and at the same time inducing glutamate release, a neurotransmitter. It is done.

The real-time measurement system of neural changes in accordance with the Ischemia using the animal model according to the present invention, the 11 vessel occlusion (VO) rat model using a variety of neurological changes (neurological changes) accompanying the induction of ischemia It is a system that measures in real time simultaneously with biological variables.

In the 11VO stroke model applied to the present invention, a cerebral ischemia is induced for 10 minutes while tracking a CCA and an ECA snare prepared by clamping by using a rubber band having a cylindrical cross section. do. After 10 minutes of induction of ischemia, the rings that blocked the blood vessels are released and reperfused again. Ischemia induction proceeds with the EEG signal disappearing completely flat and confirming a marked decrease in CBF.

1 is an overall block diagram of a system for real-time measurement of nerve changes according to Ischemia using an animal model according to the present invention.

As shown in Figure 1, the real-time measurement system of the nerve changes according to the Iskemia using the animal model according to the present invention is an apparatus for confirming the induction of Ischemia to measure the bio-parameters for confirming the induction of the Ischemia in 11VO rat model (30), Ischemia maintenance optimizer (40), which maintains a constant temperature-related bioparameter for optimization of ischemia-induced experiments in the 11VO rat model (40), a representative neuron caused by ischemia induction in the 11VO rat model The neurotransmitter measuring device 50 for measuring the amount of change in glutamate as a delivery material, and the data analysis device 60 for receiving and storing data measured from the devices 30, 40, and 50, and analyzing the data using the same. It is made to include.

The animal model according to the invention is a 11VO rat (10) model. In order to measure the glutamate level in the interstitial space of the rat in real time, it is possible to realize the forebrain ischemia as perfectly as possible. Along with ischemia, a new rat model is needed that can withstand reperfusion.

To meet this, the 11VO model is applied as an animal model that allows minimal modifications in the physiological conditions of animals and is hemodynamically stable while maintaining self-breathing life and at the same time inducing the release of glutamate, a neurotransmitter. Through this 11VO model it is possible to measure in real time changes in EEG, EEG blood pressure, blood pressure, temperature and neurotransmitters.

The apparatus for confirming the induction of escapia 30 is a device for measuring the biological parameters of the animal for estimating the cause of estimia of the animal 10 (rat) used as a model. The reason for measuring the bio-parameters as described above is to check in real time whether the animal used as the experimental model is normally in the state of Ischemia.

The escapia induction confirmation device 30 includes a means for measuring an electroencephalogram (EEG) of an animal, a means for measuring a cerebral blood flow (CBF) change of an animal, and an animal blood pressure (BP: blood). pressure means).

The electroencephalogram measuring means 31 for measuring the electroencephalogram change of the animal performs a function of measuring the electroencephalogram change of the animal which is an experimental model. Referring to the method for measuring the changes in the EEG of the animal of the experimental model as follows.

Figure 2 is a schematic diagram showing the respective craniotomy 20 of the rat model for the real-time measurement used in the present invention and the respective positions for the measurement of the biological variables.

Coronal suture (“a” in FIG. 1) of the craniotomy of the rat model makes two grooves 23 ′ at 4 mm bidirectionally from the midline and from these grooves 23 ′. Two grooves 23 "are made 4 mm backwards. Then, a screw screw is inserted into each of the four grooves 23 'and 23" in total.

The ground was made by fixing the electrode cream on both ears of the experimental model rat, and the two electrodes and the four screws were connected by wires, respectively.

The EEG values measured and configured as described above are input through the four channels (four screws), and the input analog signals are converted into digital values through a D / A converter and output. The output digital value is stored in the data analysis device 60.

The electroencephalogram values measured as described above will be drastically reduced compared to the previous time when the escapia is induced. In other words, it can be seen that the escapia is generated by confirming a rapid decrease in the measured EEG value.

The biological parameter measured along with the EEG value is the brain blood flow. This is measured by the brain blood flow measuring means 33. The brain blood flow measurement method will also be described with reference to FIG. 2.

A pair of 1.5 mm diameter grooves 21 are formed at a position 3 mm posterior from the coronal suture (“a” in FIG. 1) and 4 mm from the midline of the craniotomy of the rat model. The blood flow measurement probe is inserted into the blood flow measurement. This cerebral blood flow also tends to decrease sharply as it enters the Eskimia state. Therefore, it can be seen that the escapia is occurring by confirming a rapid decrease in the measured brain blood flow. When the animal model is an anesthetized animal, it is essential to measure the brain blood flow in order to confirm the induction of escapia.

Along with the changes in EEG and changes in brain blood flow, the biometric parameters measured for estimating the induction of eismia are the blood pressure of the animal. The animal blood pressure is made by the blood pressure measuring means 35.

For measurement of blood pressure and pulse rate using the arterial line, the right femoral artery was detached and then inserted into the artery using a 0.2 mm diameter tube and fixed using 4.0 black silk. Blood pressure is measured by connecting an A-line attached to an animal model with a pressure transducer (Harvard apparatus).

It is known that blood pressure rises temporarily when an intense ischemia stroke is triggered. Therefore, it is necessary to continuously measure the change in blood pressure to confirm the increase in blood pressure when ischemia is induced at 11VO.

The values measured by the EEG measuring unit 31, the brain blood flow measuring unit 33, and the blood pressure measuring unit 35 are converted into digital values through an A / D converter, and then stored in the data analyzing apparatus 60. It is analyzed as needed.

The above-mentioned escapia induction confirmation device 30 is a device for confirming whether the animal's escapia is well made, and in addition to maintaining the survival of the animal and optimizing the real-time measurement of nerve changes according to the escapia. In order to measure and maintain the biological parameters of the animal should also be performed. This is done in the Ischemia maintenance optimization device 40.

The escapia maintenance optimum device 40 includes body temperature measurement and maintenance means 41 and stroke volume for maintaining the body temperature of the animal at a predetermined temperature in order to prevent a sudden drop in body temperature of the animal in the escapia state. It consists of a brain temperature measurement and maintenance means 43 for maintaining the temperature of the brain at a predetermined temperature to maximize the temperature.

The body temperature measuring and maintaining means 41 adjusts the body temperature of the animal to be maintained at a predetermined temperature (about 37.5 ° C.) while measuring the temperature of the animal model in real time using a Homeothermic Blanket Control Unit (Harvard apparatus). . The reason for maintaining the body temperature of the animal in this way is to maintain the survival of the experimental animal model by preventing a rapid decrease in body temperature due to damage such as Eskemia.

In addition, the brain temperature measuring and maintaining means 43 prevents the protection effect as the temperature decreases during cerebral ischemia, thereby maximizing the stroke volume. This maximizes glutamate release and maintains the optimum temperature of the enzyme used for glutamate detection to maximize the reaction.

As shown in FIG. 2, the brain temperature forms a groove 25 using a diamond drill 1 mm in front of the coronal suture and 4 mm from the midline. The thermocouple is contacted and a temperature controller is used to maintain the brain temperature at a defined temperature (about 37.5 ° C).

The biological parameters (eg, electroencephalogram, brain blood flow, blood pressure, body temperature, and brain temperature) are measured to measure the amount of change in neurotransmitter released from the animal's Essemia state while maintaining an optimal state. This is done through the neurotransmitter measuring device 50.

In the 11VO rat model to be applied to the present invention, a representative neuronal change caused by estimia induction is a change in glutamate, a neurotransmitter with excitatory toxicity. In order to measure this, the neurotransmitter measuring apparatus 50 measures the release amount of glutamate using a microdialysis biosensor or a CNT probe.

The method for measuring the glutamate is stabilized for at least 1 hour in a constant temperature cell containing a buffer solution that is pre-calibrated micro-dialasis biosensor is set to 37.5 ℃. Then, as shown in FIG. 2, the stabilized microdialysis biosensor was slowly added to the 11VO rat model in which a hole 27 was drilled using a diamond drill 1 mm in front of the coronal suture and 4 mm from the midline. Insert it. The data is collected by operating the biosensor and the measuring device through a cable. Alternatively, insert the enzyme-fixed CNT probe to collect data. Here, the CNT probe uses the sensor described in Patent Application 10-2007-0032581.

When ischemia is induced and blood supply to the brain stops, the excitatory secretion of glutamate is promoted, resulting in cell apotosis. Therefore, real-time observations of glutamate changes can be helpful in assessing brain injury that may be involved during or after surgery, as well as to determine the effects of multiple drugs in real time. Therefore, the 11VO animal model measures and observes the neurological changes called glutamate release in real time according to ischemia.

The data measured from the devices 30, 40, 50 are all converted into digital values via D / A converters. These digitally converted values are transferred to the data analysis device 60 and stored. Then, the data analysis device 60 may analyze the relationship between the biological parameters and the amount of change of the neurotransmitter related thereto using the data. In addition, the bio parameter values and the amount of change in the neurotransmitter can be monitored in real time.

1 is a block diagram of a system for real-time measurement of nerve changes according to ischemia using an animal model applied to the present invention.

FIG. 2 is a schematic diagram showing respective positions for craniiotomy and bioparametric measurements of a rat model for real time measurement used in the present invention.

Claims (5)

In the real-time nervous system measurement system according to the Iskemia of the animal model, Ischemia-induced identification device for measuring the biological parameters of the animal to confirm the induction of the animal (ischemia); An Ischemia maintenance optimum device for measuring and maintaining the biometric parameters of the animal to maintain survival of the animal according to the Ischemia; Neurotransmitter measuring device for measuring the amount of change in the neurotransmitter released in the escape state of the animal; Real-time nervous system measurement system according to Iskemia of the animal model, characterized in that comprises a data analysis device for receiving and storing and analyzing the data from the devices. The method according to claim 1, The apparatus for confirming the induction of escapia comprises an electroencephalogram measuring means for measuring changes in the brain wave of an animal, a blood flow measuring means for measuring blood flow changes in the animal brain, and a blood pressure measuring means for measuring blood pressure changes in the animal. Real-time nervous system measurement system according to Iskemia of the animal model. The method according to claim 2, The measured electroencephalogram, brain blood flow, and blood pressure values are converted into digital values through an A / D converter, and then stored in the data analysis device. The method according to claim 1, The apparatus for maintaining Eskemia is a device for measuring and maintaining body temperature at a predetermined temperature and a stroke volume to maximize stroke volume in order to prevent a sudden decrease in body temperature of the animal in an escape state. Real-time nervous system measurement system according to Eskemia of the animal model, characterized in that it comprises a means for measuring and maintaining the temperature of the brain to maintain the temperature at a predetermined temperature. The method according to claim 1, The apparatus for measuring neurotransmitter is a real-time nervous system measuring system according to Eskemia of an animal model, characterized in that for measuring the amount of release of glutamate using a microdialysis biosensor or a CNT probe.

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