KR102179032B1 - Biomarkers for Distinguishing GCI and TBI - Google Patents

Biomarkers for Distinguishing GCI and TBI Download PDF

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KR102179032B1
KR102179032B1 KR1020190116174A KR20190116174A KR102179032B1 KR 102179032 B1 KR102179032 B1 KR 102179032B1 KR 1020190116174 A KR1020190116174 A KR 1020190116174A KR 20190116174 A KR20190116174 A KR 20190116174A KR 102179032 B1 KR102179032 B1 KR 102179032B1
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송동근
서상원
정전섭
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한림대학교 산학협력단
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Abstract

The present invention uses LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 as biomarkers to distinguish between global cerebral ischemia and traumatic brain injury. The present invention provides the biomarkers for distinguishing between global cerebral ischemia and traumatic brain injury, so that major acute central nervous system diseases can be appropriately diagnosed and treated.

Description

급성허혈성뇌손상 및 급성외상성뇌손상을 구별하기 위한 바이오마커 및 이의 이용{Biomarkers for Distinguishing GCI and TBI}Biomarkers for Distinguishing Acute Ischemic Brain Injury and Acute Traumatic Brain Injury and Their Use{Biomarkers for Distinguishing GCI and TBI}

본 발명은 급성허혈성뇌손상 및 급성외상성뇌손상을 구별하기 위한 LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59을 바이오마커로 이용하는 것이다. 그리고 본 발명은 급성허혈성뇌손상 및 급성외상성뇌손상을 구별하기 위한 바이오마커를 제공함으로써, 주요 급성 중추신경계 질환들이 적절히 진단되고 치료할 수 있도록 활용될 수 있다.The present invention uses LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 as biomarkers to distinguish between acute ischemic brain injury and acute traumatic brain injury. In addition, the present invention provides a biomarker for distinguishing between acute ischemic brain injury and acute traumatic brain injury, so that major acute central nervous system diseases can be properly diagnosed and treated.

뇌 세포들의 파괴 또는 퇴화를 유발하는 뇌손상(brain injury)은 비교적 빈번하고 광범위하게 일어나므로 의학적으로는 매우 다양한 종류의 진단방법이 요구된다. 뇌손상은 광범위한 내적 및 외적 요인들에 의해 일어나는데, 가장 많은 수의 손상을 포함하는 흔한 유형은 외부 원인으로부터의 물리적인 외상 또는 두부 손상에 따른 외상성 뇌손상(TBI)이며, 뇌혈류가 차단되어 발생하는 허혈성 뇌손상 등이 있다.Brain injury that causes destruction or degeneration of brain cells is relatively frequent and widespread, so medically, a wide variety of diagnostic methods are required. Brain injury is caused by a wide range of internal and external factors, and the most common type, including the greatest number of injuries, is physical trauma from external causes or traumatic brain injury (TBI) resulting from head injury, which is caused by blocked blood flow to the brain. Such as ischemic brain injury.

뇌손상은 그 발병 원인이 아직까지 명확히 밝혀지지 않았으나, 장기적 저산소증, 알콜을 포함한 기형 발생물질들에 의한 중독, 감염 및 신경학적 질병 등이 원인이 되는 것으로 알려져 있다. 이 외에도 뇌손상의 흔한 원인으로는 외상적 뇌손상, 뇌졸중, 동맥류, 수술, 기타 신경학적 장애, 및 중금속 중독 등과 같은 생리적 외상이 있을 수 있다. 허혈성 손상이 발생하면 뇌는 급성 염증과정과 만성 염증과정으로 반응하며 이는 주로 소교세포의 활성화, 염증 매개물질들의 생성, 호중구, T 세포, 단핵구/대식세포(monocyte/macrophage) 등의 염증성 세포들이 침윤한다. 일차적으로 수 시간 이내에 말초 혈액으로부터 백혈구(leukocytes)가 뇌 실질 부위로 침입하게되며 이때 일차적으로 침윤하는 세포가 호중구 (neutrophil)이다 (Jin et al. J Leukoc Biol. 87(5):779-789,2010). 이러한 말초 호중구는 수 시간 이내에 뇌 조직으로 침윤되고 혈관손상의 유도로 인한 BBB(blood-brainbarrier) 파괴, 기타 염증 세포의 이차 침입 및 뇌 조직의 염증 반응 개시에 중요한 역할을 수행하는 것으로 보고되었다 (Kitz R et al., J Endotoxin Res. 12(6):367-374, 2006; Scholz M et al. Med Res Rev. 27(3):401-416, 2007; Jin et al. J Leukoc Biol. 87(5):779-789, 2010).The cause of brain injury has not yet been clearly identified, but it is known to be caused by prolonged hypoxia, poisoning by teratogenic substances including alcohol, infection and neurological diseases. Other common causes of brain injury may include traumatic brain injury, stroke, aneurysm, surgery, other neurological disorders, and physiological trauma such as heavy metal poisoning. When ischemic injury occurs, the brain reacts with an acute and chronic inflammatory process, which mainly activates microglial cells, produces inflammatory mediators, and infiltrates inflammatory cells such as neutrophils, T cells, and monocytes/macrophages. do. Primarily, leukocytes from peripheral blood enter the brain parenchyma within a few hours, and the primary infiltrating cells are neutrophils (Jin et al. J Leukoc Biol. 87(5):779-789, 2010). These peripheral neutrophils infiltrate into brain tissue within a few hours and have been reported to play an important role in the destruction of blood-brainbarrier (BBB) due to induction of blood vessel damage, secondary invasion of other inflammatory cells, and initiation of inflammatory responses in brain tissue (Kitz R et al., J Endotoxin Res. 12(6):367-374, 2006; Scholz M et al. Med Res Rev. 27(3):401-416, 2007; Jin et al. J Leukoc Biol. 87( 5):779-789, 2010).

한편, Specialized pro-resolving mediators (SPMs)는 주로 염증으로 부터 조직을 회복시키는 작용을 하는 생물활성 지질로서, arachidonic acid 으로부터 합성되는 lipoxin A4 (LXA4), eicosapentaenoic acid (EPA) 으로부터 합성되는 resolvin Es (RvEs), docosahexaenoic acid (DHA) 으로부터 합성되는 resolvin Ds (RvDs) 등을 포함한다. 이들 SPMs은 경동맥 질환 (Bazan et al., 2017), 폐렴 (Dalli et al., 2015), 2형 당뇨 (Barden et al., 2018), 만성 특발성 두드러기 (Dilek et al., 2018), 및 전자간증 (Perucci et al., 2016) 환자의 혈액내에서 그 농도가 증가해 있음이 보고되어있다. 그러나, 급성허혈성뇌손상 및 급성외상성뇌손상과 같은 급성 뇌손상시에 그 혈액내 SPMs의 변화는 보고되어 있지 않다. On the other hand, Specialized pro-resolving mediators (SPMs) are bioactive lipids that mainly act to restore tissue from inflammation, and resolvin Es (RvEs) synthesized from lipoxin A4 (LXA4) synthesized from arachidonic acid and eicosapentaenoic acid (EPA). ), resolvin Ds (RvDs) synthesized from docosahexaenoic acid (DHA), and the like. These SPMs are carotid artery disease (Bazan et al., 2017), pneumonia (Dalli et al., 2015), type 2 diabetes (Barden et al., 2018), chronic idiopathic urticaria (Dilek et al., 2018), and the former. It has been reported that the concentration in the blood of patients with testimony (Perucci et al., 2016) is increased. However, changes in blood SPMs during acute brain injury such as acute ischemic brain injury and acute traumatic brain injury have not been reported.

한편, CD59은 보체 조절 단백 (complement regulatory protein)의 하나로서 (Kimberley et al., 2006), 급성 심근 경색 환자 (acute myocardial infarction patients) 에서 그 혈장내 농도가 증가해 있음이 보고된바 있다(Vakeva et al., 2000). 그러나, 급성허혈성뇌손상 및 급성외상성뇌손상에서는 혈액내 CD59의 농도변화가 보고된 바 없다. 따라서, 본 발명자는 급성허혈성뇌손상 및 급성외상성뇌손상 모델 동물에서 혈액내 CD59 농도를 측정하여 각 뇌손상을 구별할 수 있는 바이오마커로 활용할 수 있는지 연구하였다. Meanwhile, CD59, as one of the complement regulatory proteins (Kimberley et al., 2006), has been reported to have an increased plasma concentration in acute myocardial infarction patients (Vakeva). et al., 2000). However, no change in blood CD59 concentration has been reported in acute ischemic brain injury and acute traumatic brain injury. Therefore, the present inventors investigated whether the CD59 concentration in the blood in acute ischemic brain injury and acute traumatic brain injury model animals can be measured and used as a biomarker that can distinguish each brain injury.

공개특허 제10-2018-0136702호Publication Patent No. 10-2018-0136702 등록특허 제10-1211280호Registered Patent No. 10-1211280 등록특허 제10-1476818호Registered Patent No. 10-1476818

본 발명은 상기한 바와 같은 종래 기술의 문제점을 해결하기 위한 것으로서,본 발명의 목적은 급성허혈성뇌손상 및 급성외상성뇌손상을 바이오마커를 이용하여 정확히 구별 진단함으로써, 주요 급성 중추신경계 질환을 적절히 치료받을 수 있도록 하기 위한 바이오마커 및 이의 이용방법을 제공하는 것이다.The present invention is to solve the problems of the prior art as described above, and an object of the present invention is to properly treat major acute central nervous system diseases by accurately distinguishing and diagnosing acute ischemic brain injury and acute traumatic brain injury using biomarkers. It is to provide a biomarker and a method of use thereof to be able to receive it.

본 발명의 해결하고자 하는 과제는 이상에서 언급된 것들에 한정되지 않으며, 언급되지 아니한 다른 해결과제들은 아래의 기재로부터 당업자에게 명확하게 이해되어 질 수 있을 것이다.The problem to be solved of the present invention is not limited to those mentioned above, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

상기한 목적을 달성하기 위하여 본 발명에 따른 급성허혈성뇌손상 및 급성외상성뇌손상을 구별하기 위한 바이오마커는 혈액에 있는 LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59 중 적어도 하나를 포함하는 것을 특징으로 한다.In order to achieve the above object, the biomarker for distinguishing acute ischemic brain injury and acute traumatic brain injury according to the present invention comprises at least one of LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 in blood. do.

또한, 본 발명에 따른 급성허혈성뇌손상 및 급성외상성뇌손상을 구별하는데 필요한 정보를 제공하는 방법은 급성허혈성뇌손상 및 급성외상성뇌손상을 구별하는 데 필요한 정보를 제공하기 위하여 혈액에 있는 LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59 중 적어도 하나를 포함하는 바이오마커의 농도를 측정하는 단계를 통하여 이루어지는 것을 특징으로 한다.In addition, the method for providing information necessary to distinguish acute ischemic brain injury and acute traumatic brain injury according to the present invention is LXA4, RvE1 in blood to provide information necessary to distinguish acute ischemic brain injury and acute traumatic brain injury. , RvE2, RvD1, RvD2 and CD59 characterized in that made through the step of measuring the concentration of a biomarker containing at least one.

보다 구체적으로, 급성허혈성뇌손상 및 급성외상성뇌손상의 발병 여부를 진단하고자 하는 대상으로부터 혈액을 얻는 단계와, 상기 시료로부터 LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59 중 적어도 하나를 포함하는 바이오마커의 농도를 각각 측정하는 단계와, 측정된 바이오마커의 농도를 비교하여 발병 여부를 결정하는 단계로 구성될 수 있다.More specifically, obtaining blood from a subject to be diagnosed with acute ischemic brain injury and acute traumatic brain injury, and a biomarker comprising at least one of LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 from the sample It may consist of a step of measuring each concentration of and determining whether or not an onset occurs by comparing the measured concentration of the biomarker.

본 발명에 따른 급성허혈성뇌손상 및 급성외상성뇌손상을 구별하기 위한 바이오마커 및 이의 이용에 따르면, LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59 중 적어도 하나를 포함하는 바이오마커의 농도를 측정함으로써 급성허혈성뇌손상 및 급성외상성뇌손상을 정확하게 구별할 수 있는 효과가 있다.According to the biomarker for distinguishing acute ischemic brain injury and acute traumatic brain injury according to the present invention and its use, acute by measuring the concentration of a biomarker comprising at least one of LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 It has the effect of accurately distinguishing between ischemic brain injury and acute traumatic brain injury.

본 발명의 효과는 이상에서 언급된 것들에 한정되지 않으며, 언급되지 아니한 다른 효과들은 아래의 기재로부터 당업자에게 명확하게 이해되어 질 수 있을 것이다.The effects of the present invention are not limited to those mentioned above, and other effects that are not mentioned will be clearly understood by those skilled in the art from the following description.

도 1은 쥐에서 GCI 및 TBI 후 혈장 LXA4의 변화를 실험한 결과이다.(*p<0.05, **p<0.01; compared to the sham group)
도 2a 내지 도 2d는 쥐에서 GCI 및 TBI 후 혈장 resolvins E1, E2, D1, and D2의 변화를 각각 실험한 결과이다.(*p<0.05, **p<0.01, ***p<0.001; compared to the sham group)
도 3은 쥐에서 GCI 및 TBI 후 혈장 CD59의 변화를 실험한 결과이다.(*p<0.05, ***p<0.001; compared to the sham group)Figure 1 shows the results of experimentation of changes in plasma LXA4 after GCI and TBI in mice (*p<0.05, **p<0.01; compared to the sham group).
2A to 2D are results of experiments on changes in plasma resolvins E1, E2, D1, and D2 after GCI and TBI in mice, respectively. (*p<0.05, **p<0.01, ***p<0.001; compared to the sham group)
Figure 3 shows the results of testing the changes in plasma CD59 after GCI and TBI in mice (*p<0.05, ***p<0.001; compared to the sham group).

이하 본 발명의 바람직한 실시예에 대하여 구체적으로 설명한다.Hereinafter, a preferred embodiment of the present invention will be described in detail.

본 발명을 설명함에 있어서, 관련된 공지기능 혹은 구성에 대한 구체적인 설명이 본 발명의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우 그 상세한 설명은 생략한다. 또한, 후술되는 용어들은 본 발명에서의 기능을 고려하여 정의된 용어들로서 이는 사용자, 운용자의 의도 또는 판례 등에 따라 달라질 수 있다. 그러므로 그 정의는 본 명세서 전반에 걸친 내용을 토대로 내려져야 할 것이다.In describing the present invention, when it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the subject matter of the present invention, a detailed description thereof will be omitted. In addition, terms to be described later are terms defined in consideration of functions in the present invention, which may vary according to intentions or precedents of users or operators. Therefore, the definition should be made based on the contents throughout this specification.

본 발명자들은 급성허혈성뇌손상 및 급성외상성뇌손상의 혈액내 biomarker의 개발을 위하여, 쥐에서 급성허혈성뇌손상(global cerebral ischemia, GCI) 및 급성외상성뇌손상(traumatic brain injury), TBI)을 유발한 다음, 경시적으로 혈액을 채취하여, LXA4, resolvin E1(RvE1), resolvin E2(RvE2), resolvin D1(RvD1), resolvin D2(RvD2), CD59의 농도의 변화를 관찰하였다.The present inventors induce acute ischemic brain injury (global cerebral ischemia, GCI) and acute traumatic brain injury (traumatic brain injury, TBI) in rats for the development of biomarkers in blood for acute ischemic brain injury and acute traumatic brain injury. Next, blood was collected over time, and changes in the concentration of LXA4, resolvin E1 (RvE1), resolvin E2 (RvE2), resolvin D1 (RvD1), resolvin D2 (RvD2), and CD59 were observed.

[실험예 1][Experimental Example 1]

급성허혈성뇌손상(global cerebral ischemia, GCI) 유도Induction of acute ischemic brain injury (global cerebral ischemia, GCI)

GCI는 bilateral common carotid artery occlusion 으로 유도하였다(Suh et al., 2008; Hong et al., 2018). GCI was induced by bilateral common carotid artery occlusion (Suh et al., 2008; Hong et al., 2018).

즉, 쥐는 isoflurane vaporizer(VetEquip)를 사용하여 isoflurane(1~2% for maintenance; 3% for induction) in a 70:30 mixture of nitrous oxide/oxygen 으로 깊게 마취하였고, 정온 담요 조절기(homeothermic blanket control unit, Harvard apparatus)를 이용하여 심부 체온(core temperature)을 36.5~37.5℃로 유지하였다. 동맥압 측정과 혈액을 채취하기 위하여 heparin을 채운 catheter를 femoral artery에 삽입하였다. 목 정중앙 절개(midline neck incision)로 양쪽 총경동맥 (common carotid artery)를 노출시킨 다음, 패색(occlusion)의 기시부에 4/O silk suture로 두른다. 뇌파기록을 위한 probe(electroencephalographic (EEG) probes)를 bilateral burr holes에 위치시킨다.That is, mice were deeply anesthetized with isoflurane (1~2% for maintenance; 3% for induction) in a 70:30 mixture of nitrous oxide/oxygen using isoflurane vaporizer (VetEquip), and a homeothermic blanket control unit, The core temperature was maintained at 36.5~37.5℃ using a Harvard apparatus. In order to measure arterial pressure and collect blood, a catheter filled with heparin was inserted into the femoral artery. Expose both common carotid arteries with a midline neck incision, then wrap with 4/O silk suture at the beginning of the occlusion. Probes for EEG recording (electroencephalographic (EEG) probes) are placed in the bilateral burr holes.

전신평균 동맥압 (mean arterial pressure, MAP)을 대퇴동맥(femoral artery)로 부터 혈액(7~10cc)을 뽑아냄으로써, 40±10 mmHg 로 감소시킨 다음, 노출된 양쪽 총경동맥을 clamp 한다.The mean arterial pressure (MAP) is reduced to 40±10 mmHg by drawing blood (7-10 cc) from the femoral artery, and then clamping both exposed common carotid arteries.

성공적인 GCI 유도는 EEG monitor 상의 isoelectricity로 확인한다. 7분간의 isoelectric EEG signal 후에, clamp를 제거하여 혈류를 회복시켜준다. Successful GCI induction is confirmed by isoelectricity on the EEG monitor. After 7 minutes of isoelectric EEG signal, remove the clamp to restore blood flow.

피부 절개부위의 suture 후에, 마취를 중단한다. 쥐가 자발 호흡을 하게되면, 37℃로 유지되는 incubator에 넣는다. 대조군은 동일한 isoflurane 마취하에서 동일한 목부위 피부절개를 받지만, 총경동맥 패색을 시행하지 않았다.After suture of the skin incision, stop anesthesia. When the rat spontaneously breathes, it is placed in an incubator maintained at 37°C. The control group received the same neck skin incision under the same isoflurane anesthesia, but did not perform common carotid artery occlusion.

[실험예 2][Experimental Example 2]

급성외상성뇌손상(traumatic brain injury, TBI) 유도Induction of acute traumatic brain injury (TBI)

실험적 TBI를 위한 controlled cortical impact (CCI) model은 아래와 같다(Choi et al., 2012, 2019). The controlled cortical impact (CCI) model for experimental TBI is as follows (Choi et al., 2012, 2019).

쥐는 isoflurane vaporizer (VetEquip)를 사용하여 isoflurane (마취유지시 1~2%; 마취유도시 3%) in a 70:30 mixture of nitrous oxide/oxygen 으로 깊게 마취하였고, stereotaxic apparatus (David Kopf Instruments, Tujunga, CA, USA) 에 위치시켰다. 휴대용 드릴 (정중앙선으로부터 옆으로 2.8 mm, 그리고 bregma 뒤쪽으로 3 mm)을 이용하여 좌반구위 약 5 mm정도로 craniotomy를 시행하였다.The rats were deeply anesthetized with isoflurane (1-2% during anesthesia maintenance; 3% when anesthesia was induced) in a 70:30 mixture of nitrous oxide/oxygen using isoflurane vaporizer (VetEquip), and a stereotaxic apparatus (David Kopf Instruments, Tujunga, CA, USA). A craniotomy was performed about 5 mm above the left hemisphere using a hand-held drill (2.8 mm laterally from the midline and 3 mm behind the bregma).

controlled cortical impact device (Leica Impact One; Leica Biosystems, Nussloch, Germany)를 이용하여, 3mm flat-tip impactor가 5m/sec의 속도로 가속되어 3mm 깊이로 충격이 되도록 하였다.Using a controlled cortical impact device (Leica Impact One; Leica Biosystems, Nussloch, Germany), a 3mm flat-tip impactor was accelerated at a speed of 5m/sec to make an impact to a depth of 3mm.

모든 쥐는 수술 도중 및 수술 후 일어나서 움직일 때까지 정온담요 조절기 (homeothermic blanket control unit, Harvard Bioscience, Holliston, MA, USA) 를 이용하여 심부 체온 36~37.5℃로 유지되도록 한다. 대조군은 개두술 (craniotomy)만 받았다.During and after surgery, all mice are kept at a core temperature of 36-37.5°C using a homeothermic blanket control unit (Harvard Bioscience, Holliston, MA, USA) until they get up and move. The control group received only craniotomy.

[실시예 1] : 혈액으로부터 LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59의 측정[Example 1]: Measurement of LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 from blood

마취 후, 안와후 정맥총(retro-orbital venous plexus)에서 혈액(1.5 ml)을 뇌손상후 6, 24, 72, and 168 h에 각각 대조준 및 실험군에서 채취한다. 각 동물의 수는 5마리로 하였다. LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59의 혈장 농도는 ELISA (MyBioSource) 키트로 측정하였으며, 정상치는 아무 처치도 하지 않은 3마리의 동물군으로부터 구하였고, 그 결과는 도 1 내지 도 3의 실험 결과와 같다.After anesthesia, blood (1.5 ml) was collected from the retro-orbital venous plexus at 6, 24, 72, and 168 h after brain injury from the control group and the experimental group, respectively. The number of each animal was set to five. Plasma concentrations of LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 were measured with an ELISA (MyBioSource) kit, and the normal value was obtained from a group of 3 animals without any treatment, and the results were obtained from the experiments of FIGS. 1 to 3 Same as the result.

도 1에 나타난 바와 같이, 혈액 내 LXA4의 농도는 GCI 후 24시간, 72시간에 유의한 증가를 보였으며, TBI 후 72시간에서는 유의한 감소를 보였다.As shown in FIG. 1, the concentration of LXA4 in the blood showed a significant increase at 24 hours and 72 hours after GCI, and a significant decrease at 72 hours after TBI.

도 2a 내지 도 2d 각각에 나타난 바와 같이, 혈액 내 RvE1, RvE2, RvD1, RvD2의 농도는 GCI 후 6시간에 모두 유의한 감소를 보였으며, 72시간에서는 RvE1, RvD1, RvD2의 농도가 유의한 감소를 보였다. 반면에, 혈액내 RvE1, RvE2, RvD1, RvD2의 농도는 TBI 후에서 유의한 변화를 보이지 않았다.2A to 2D, the concentrations of RvE1, RvE2, RvD1, and RvD2 in the blood all showed a significant decrease at 6 hours after GCI, and at 72 hours, the concentrations of RvE1, RvD1, and RvD2 were significantly decreased. Showed. On the other hand, the concentrations of RvE1, RvE2, RvD1, and RvD2 in the blood did not show any significant change after TBI.

도 3에 나타난 바와 같이, 혈액 내 CD59의 농도는 GCI 후 6시간 및 24시간에 유의한 증가를 보였으며, TBI 후 6시간에서는 유의한 감소를 보였다.As shown in FIG. 3, the concentration of CD59 in the blood showed a significant increase at 6 hours and 24 hours after GCI, and a significant decrease at 6 hours after TBI.

이상의 결과들을 통해 혈액 내 LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59들이 급성허혈성뇌손상(global cerebral ischemia, GCI)과 급성외상성뇌손상(traumatic brain injury)를 감별 진단을 하는데 유용한 바이오마커로 이용될 수 있음을 확인할 수 있었다.Based on the above results, LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 in the blood can be used as useful biomarkers for differential diagnosis of global cerebral ischemia (GCI) and acute traumatic brain injury. It could be confirmed that it can be.

이상에서 설명된 본 발명은 예시적인 것에 불과하며, 본 발명이 속한 기술분야의 통상의 지식을 가진 자라면 이로부터 다양한 변형 및 균등한 타 실시예가 가능하다는 점을 잘 알 수 있을 것이다. 그러므로 본 발명은 상기의 상세한 설명에서 언급되는 형태로만 한정되는 것은 아님을 잘 이해할 수 있을 것이다. 따라서 본 발명의 진정한 기술적 보호 범위는 첨부된 특허청구범위의 기술적 사상에 의해 정해져야 할 것이다. 또한, 본 발명은 첨부된 청구범위에 의해 정의되는 본 발명의 정신과 그 범위 내에 있는 모든 변형물과 균등물 및 대체물을 포함하는 것으로 이해되어야 한다.The present invention described above is merely exemplary, and those of ordinary skill in the art to which the present invention pertains will appreciate that various modifications and other equivalent embodiments are possible. Therefore, it will be appreciated that the present invention is not limited to the form mentioned in the detailed description above. Therefore, the true technical protection scope of the present invention should be determined by the technical spirit of the appended claims. In addition, the present invention is to be understood as including the spirit of the present invention as defined by the appended claims and all modifications, equivalents and substitutes within the scope thereof.

Claims (2)

급성허혈성뇌손상 및 급성외상성뇌손상을 구별하는 데 필요한 정보를 제공하기 위하여 혈액에 있는 LXA4, RvE1, RvE2, RvD1, RvD2 및 CD59로 이루어지는 바이오마커들의 농도를 측정하는 단계를 통하여 급성허혈성뇌손상 및 급성외상성뇌손상을 구별하는데 필요한 정보를 제공하는 방법.
Acute ischemic brain injury and acute ischemic brain injury through the step of measuring the concentration of biomarkers consisting of LXA4, RvE1, RvE2, RvD1, RvD2 and CD59 in the blood in order to provide necessary information to distinguish between acute ischemic brain injury and acute traumatic brain injury. A method of providing the information necessary to distinguish acute traumatic brain injury.
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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060024749A1 (en) * 2001-06-27 2006-02-02 Dambinova Svetlana A Rapid multiple panel of biomarkers in laboratory blood tests for TIA/stroke
KR20110107981A (en) * 2010-03-26 2011-10-05 가톨릭대학교 산학협력단 Novel markers for diagnosing brain disease caused by brain injury and use thereof
KR20130043104A (en) * 2010-04-06 2013-04-29 카리스 라이프 사이언스 룩셈부르크 홀딩스 Circulating biomarkers for disease
KR101476818B1 (en) 2012-01-27 2014-12-29 의료법인 성광의료재단 Biomarker Indicative of Reduce or Alleviation of Symptom of Brain Injury
US20150018234A1 (en) * 2012-01-07 2015-01-15 The Regents Of The University Of California Biomarkers for diagnosing ischemia
KR20170105184A (en) * 2016-03-09 2017-09-19 아주대학교산학협력단 Biomarker composition for predicting prognosis of intrahepatic cholangiocarcinoma comprising 794 gene set
EP3336551A1 (en) * 2008-08-11 2018-06-20 Banyan Biomarkers, Inc. Biomarker detection process and assay of neurological condition
KR20180136702A (en) 2017-06-15 2018-12-26 가톨릭대학교 산학협력단 Use of the autofluorescence, as a marker for diagnosing brain damages

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060024749A1 (en) * 2001-06-27 2006-02-02 Dambinova Svetlana A Rapid multiple panel of biomarkers in laboratory blood tests for TIA/stroke
EP3336551A1 (en) * 2008-08-11 2018-06-20 Banyan Biomarkers, Inc. Biomarker detection process and assay of neurological condition
KR20110107981A (en) * 2010-03-26 2011-10-05 가톨릭대학교 산학협력단 Novel markers for diagnosing brain disease caused by brain injury and use thereof
KR101211280B1 (en) 2010-03-26 2012-12-11 가톨릭대학교 산학협력단 Novel markers for diagnosing brain disease caused by brain injury and use thereof
KR20130043104A (en) * 2010-04-06 2013-04-29 카리스 라이프 사이언스 룩셈부르크 홀딩스 Circulating biomarkers for disease
US20150018234A1 (en) * 2012-01-07 2015-01-15 The Regents Of The University Of California Biomarkers for diagnosing ischemia
KR101476818B1 (en) 2012-01-27 2014-12-29 의료법인 성광의료재단 Biomarker Indicative of Reduce or Alleviation of Symptom of Brain Injury
KR20170105184A (en) * 2016-03-09 2017-09-19 아주대학교산학협력단 Biomarker composition for predicting prognosis of intrahepatic cholangiocarcinoma comprising 794 gene set
KR20180136702A (en) 2017-06-15 2018-12-26 가톨릭대학교 산학협력단 Use of the autofluorescence, as a marker for diagnosing brain damages

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
Brain Research (2012) 1481:49-58 *
Frontiers in Neurology (2018) 9:617 *
Molecular Brain (2018) 11:9 *
Neural Regeneration Research (2016) 11(1):77-78 *
The Korean Journal of Physiology & Pharmacology (2020) 24(2):165-171 *

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