JP2018124219A - Method of aiding determination of risk of cardiovascular and other diseases - Google Patents
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 11
- 230000002526 effect on cardiovascular system Effects 0.000 title 1
- 208000006011 Stroke Diseases 0.000 claims abstract description 33
- 210000004369 blood Anatomy 0.000 claims abstract description 25
- 239000008280 blood Substances 0.000 claims abstract description 25
- 208000029078 coronary artery disease Diseases 0.000 claims abstract description 25
- 208000024172 Cardiovascular disease Diseases 0.000 claims abstract description 13
- 238000003745 diagnosis Methods 0.000 claims description 7
- 108010007622 LDL Lipoproteins Proteins 0.000 abstract description 19
- 102000007330 LDL Lipoproteins Human genes 0.000 abstract description 19
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract 1
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- 102000004895 Lipoproteins Human genes 0.000 description 6
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- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 6
- 108010010234 HDL Lipoproteins Proteins 0.000 description 4
- 102000015779 HDL Lipoproteins Human genes 0.000 description 4
- 108010028554 LDL Cholesterol Proteins 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 235000012000 cholesterol Nutrition 0.000 description 3
- 208000019423 liver disease Diseases 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 206010003210 Arteriosclerosis Diseases 0.000 description 2
- 108010004103 Chylomicrons Proteins 0.000 description 2
- 108010046315 IDL Lipoproteins Proteins 0.000 description 2
- 108010062497 VLDL Lipoproteins Proteins 0.000 description 2
- 208000011775 arteriosclerosis disease Diseases 0.000 description 2
- 208000037998 chronic venous disease Diseases 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 208000031225 myocardial ischemia Diseases 0.000 description 2
- 108010071584 oxidized low density lipoprotein Proteins 0.000 description 2
- 102000006410 Apoproteins Human genes 0.000 description 1
- 108010083590 Apoproteins Proteins 0.000 description 1
- 238000008214 LDL Cholesterol Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
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- -1 triglyceride peroxide Chemical class 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
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Abstract
Description
本発明は、循環器疾患、冠動脈心疾患又は脳卒中のリスクの判断を補助する方法、及び脳卒中の診断を補助する方法に関する。 The present invention relates to a method for assisting in the determination of the risk of cardiovascular disease, coronary heart disease or stroke, and a method for assisting in the diagnosis of stroke.
血清・血漿中の脂質の主な成分は、コレステロール、トリグリセリド、リン脂質等であり、これら血中脂質はアポ蛋白と結合し、リポ蛋白として血液中を循環する。これらリポ蛋白は密度の差により、カイロミクロン(CM)、超低密度リポ蛋白（VLDL）、中間密度リポ蛋白（IDL）、低密度リポ蛋白（以下、LDLと称することがある）、高密度リポ蛋白（以下、HDLと称することがある）等に分類される。これらリポ蛋白のうち、LDLは肝臓から各組織へのコレステロールの主たる運搬体であり、LDLコレステロール（以下、LDL-Cと称することがある）の増加は動脈硬化発生と密接な関係があるといわれている。このことから、LDL-Cは、動脈硬化症、虚血性心疾患（冠動脈疾患）等の危険因子と考えられ、LDL-Cの含有量を知ることは、これら疾患の診断・治療および予防において重要な指標とされている。その一方で、血液中のLDL-Cが正常範囲であっても虚血性心疾患等を発症する例も数多く認められ、最近ではLDL粒子の質の変化にも注目するようになってきた。 The main components of lipids in serum and plasma are cholesterol, triglycerides, phospholipids, etc., and these blood lipids bind to apoproteins and circulate in the blood as lipoproteins. These lipoproteins are different in density from chylomicron (CM), very low density lipoprotein (VLDL), intermediate density lipoprotein (IDL), low density lipoprotein (hereinafter sometimes referred to as LDL), high density lipoprotein. It is classified into protein (hereinafter sometimes referred to as HDL). Among these lipoproteins, LDL is the main transporter of cholesterol from the liver to each tissue, and the increase in LDL cholesterol (hereinafter sometimes referred to as LDL-C) is said to be closely related to the development of arteriosclerosis. ing. Therefore, LDL-C is considered to be a risk factor for arteriosclerosis, ischemic heart disease (coronary artery disease), etc. Knowing the content of LDL-C is important in the diagnosis, treatment and prevention of these diseases It is considered as an index. On the other hand, there are many cases in which ischemic heart disease or the like develops even when LDL-C in the blood is in the normal range, and recently, attention has been paid to changes in the quality of LDL particles.
トリグリセリドを多く含むLDL（以下、TG-rich LDLと称することがある）は、コレステロールの含有量が高い正常なＬＤＬとは性状が異なるリポタンパク質で、肝疾患患者血液中に多く見られ、肝疾患の進行に伴い血液中濃度は増加し、肝疾患の末期では血液中に存在するリポタンパク質の大部分を占めるとの報告がある。また、TG-rich LDLはマクロファージを泡沫化するが、このTG-rich LDLによるマクロファージ泡沫化率と酸化LDLの一種であるマロンジアルデヒド修飾LDLの血清中濃度とが正比例すること、健常者血液中には過酸化トリグリセリドがほとんど検出されないのに対し、肝疾患患者血液中には過酸化トリグリセリドの著しい増加が見られること、などの報告により、LDL中のトリグリセリドは酸化LDLとの関連も深いと考えられている。冠動脈心疾患（Coronary Heart Disease：CHD）を起こしている患者と健常人のLDL中のトリグリセリド（以下、LDL-TGと称することがある）を比較した場合、冠動脈心疾患患者では血中のLDL-TGが増加することが報告されている（非特許文献１）。しかし現在冠動脈疾患を発症していない健常人の中でもLDL-TGが高い人が将来冠動脈心疾患発症のリスクが高くなることを示すエビデンスは存在しない。 LDL containing a large amount of triglycerides (hereinafter sometimes referred to as TG-rich LDL) is a lipoprotein that is different from normal LDL with a high cholesterol content and is often found in the blood of patients with liver disease. It has been reported that the blood concentration increases with the progression of the disease and accounts for the majority of lipoproteins present in the blood at the end of liver disease. In addition, TG-rich LDL foams macrophages, but the macrophage foaming rate by TG-rich LDL is directly proportional to the serum concentration of malondialdehyde-modified LDL, which is a kind of oxidized LDL. According to reports that triglyceride in LDL is closely related to oxidized LDL, there is a significant increase in triglyceride peroxide in the blood of patients with liver disease. It has been. When comparing triglycerides in LDL (hereinafter sometimes referred to as LDL-TG) in patients with coronary heart disease (CHD) and healthy individuals, LDL- in the blood in patients with coronary heart disease It has been reported that TG increases (Non-patent Document 1). However, there is no evidence that healthy individuals who currently do not develop coronary artery disease have high LDL-TG risk of developing future coronary heart disease.
一方、脳卒中に関してはLDL-TGとの関連性を示すエビデンスが存在せず、脳卒中を起こしている患者の診断に使用できるかどうか、またはLDL-TG高値の被験者が将来脳卒中を発症するリスクが高まるかどうかは明らかとなっていない。 On the other hand, there is no evidence of an association with LDL-TG for stroke, whether it can be used to diagnose patients with stroke, or subjects with high LDL-TG are at increased risk of developing a stroke in the future Whether it is not clear.
LDL-TGの定量方法としては、分画分離とトリグリセリド定量の２つの段階操作を組み合わせて求める方法がある。分画分離操作は、超遠心法、電気泳動法、高速液体クロマトグラフィーを利用した方法等があり、定量法としては、たとえば臨床検査の場で用いられている自動分析装置を用いてトリグリセリド測定用試薬により定量を行う方法等がある。この両者を組み合わせて、LDL-TGの定量を行うことができるが、これらはLDLとLDL以外のリポ蛋白とを完全に分離する前処理工程と、測定を行う工程の二段階で行われるため、操作が煩雑で時間がかかる。また分離方法によっては、分離した試料そのものの回収が困難であったり、定量的に回収することが困難であったり、定量的に回収することができる方法でも、作業に熟練が必要であったり、特別な機器が必要であったりする。これらは費用も高額となり、簡便性や経済性において普及しにくい方法である。 As a method for quantifying LDL-TG, there is a method of determining by combining two step operations of fractional separation and triglyceride quantification. The fraction separation operation includes methods such as ultracentrifugation, electrophoresis, and high-performance liquid chromatography. The quantitative method is, for example, for triglyceride measurement using an automatic analyzer used in the clinical laboratory. There is a method of quantifying with a reagent. By combining these two, LDL-TG can be quantified, but since these are performed in two steps, a pretreatment step that completely separates LDL and lipoproteins other than LDL, and a measurement step, The operation is complicated and takes time. Depending on the separation method, it is difficult to collect the separated sample itself, it is difficult to collect quantitatively, even in a method that can be quantitatively collected, skill is required for work, Special equipment is necessary. These methods are expensive, and are difficult to spread in terms of simplicity and economy.
これらの問題点を解消する方法で、分画操作を行うことなく自動分析装置等で測定可能な方法としては、第１工程でLDL以外の全てのリポ蛋白中のトリグリセリドを除去した後、第２工程で残ったLDL中のトリグリセリドを測定する方法（特許文献１、２）や、第１工程で（遊離型グリセロールおよび）HDL中のトリグリセリドを除去した後、第２工程でLDL中のトリグリセリドのみを測定する方法（特許文献３）が知られている。 As a method for solving these problems, which can be measured by an automatic analyzer without performing a fractionation operation, after removing triglycerides in all lipoproteins other than LDL in the first step, Method of measuring triglycerides in LDL remaining in the process (Patent Documents 1 and 2) or removing triglycerides in HDL (free glycerol and HDL) in the first process, then only triglycerides in LDL in the second process A measuring method (Patent Document 3) is known.
本発明の目的は、循環器疾患、冠動脈心疾患又は脳卒中のリスクの判断を補助する方法を提供することである。さらに本発明の目的は、脳卒中の診断を補助する方法を提供することである。 It is an object of the present invention to provide a method that assists in determining the risk of cardiovascular disease, coronary heart disease or stroke. It is a further object of the present invention to provide a method that assists in the diagnosis of stroke.
本願発明者らは鋭意研究の結果、循環器疾患、冠動脈心疾患又は脳卒中を発症していない健常者において、血中LDL-TG値が高い場合には、将来的に循環器疾患、冠動脈心疾患又は脳卒中を発症する可能性が高くなることを見出し、また、脳卒中を発症している患者において、血中LDL-TG値が高いことを見出し、本発明を完成した。 As a result of diligent research, the inventors of the present application have found that, in healthy subjects who have not developed cardiovascular disease, coronary heart disease or stroke, if blood LDL-TG level is high, cardiovascular disease, coronary heart disease in the future Alternatively, the present inventors have found that the possibility of developing a stroke is high, and found that the blood LDL-TG level is high in patients who have developed a stroke, thereby completing the present invention.
すなわち、本発明は、以下のものを提供する。
(1) 生体から分離された血液中のLDL-TG値を測定することを含み、測定されたLDL-TG値が高いことが、循環器疾患、冠動脈心疾患又は脳卒中のリスクが高いことを示す、循環器疾患、冠動脈心疾患又は脳卒中のリスクの判断を補助する方法。
(2) 生体から分離された血液中のLDL-TG値を測定することを含み、測定されたLDL-TG値が高いことが、脳卒中を発症している可能性が高いことを示す、脳卒中の診断を補助する方法。
That is, the present invention provides the following.
(1) Including the measurement of LDL-TG level in blood separated from the living body, a high measured LDL-TG level indicates a high risk of cardiovascular disease, coronary heart disease or stroke A method to assist in the determination of the risk of cardiovascular disease, coronary heart disease or stroke.
(2) It includes measuring the LDL-TG level in blood separated from the living body, and a high measured LDL-TG value indicates a high possibility of developing a stroke. A method to aid diagnosis.
本発明により、循環器疾患、冠動脈心疾患又は脳卒中を発症していない段階において、将来的にこれらの疾患を発症するリスクの高低を的確に判断する方法が初めて提供された。リスクが高いと判断された場合には、生活習慣を改めたり、薬を服用したりすることにより、リスクの低減に努めることが可能となり、これらの疾患の発症を予防することが可能になる。 The present invention provides for the first time a method for accurately determining the risk of developing these diseases in the future in the stage where no cardiovascular disease, coronary heart disease or stroke has developed. When it is determined that the risk is high, it is possible to try to reduce the risk by changing lifestyle habits or taking medicine, and it is possible to prevent the onset of these diseases.
また、本発明により、脳卒中を迅速、的確に診断することを補助することが可能になった。したがって、脳卒中発症の初期において、迅速な処置を行うことが可能となり、脳卒中が原因の死亡を防ぐことができる。 In addition, the present invention can assist in quickly and accurately diagnosing a stroke. Therefore, rapid treatment can be performed in the early stage of stroke onset, and death due to stroke can be prevented.
本発明の方法では、血液中のLDL-TG値を測定する。血液試料としては、血清又は血漿を用いることが、測定を行いやすいので好ましい。 In the method of the present invention, the LDL-TG level in blood is measured. As the blood sample, it is preferable to use serum or plasma because measurement is easy.
血液中のLDL-TG値を測定する方法自体は周知（例えば上記特許文献１〜３）であり、そのための試薬も市販されているので、市販の試薬を用いて容易に血中LDL-TGを定量することができる。 The method itself for measuring the LDL-TG value in blood is well known (for example, Patent Documents 1 to 3 above), and reagents for that purpose are also commercially available. Therefore, LDL-TG in blood can be easily determined using commercially available reagents. It can be quantified.
循環器疾患、冠動脈心疾患又は脳卒中を発症していない健常者において、血中LDL-TG値が高いほど、将来的にこれらの疾患を発症する可能性が高くなる。したがって、測定された血中LDL-TG値に基づいて将来的にこれらの疾患を発症するリスクの判断を補助することができる。血中LDL-TG値が高いほど、将来的にこれらの疾患を発症する可能性が高くなるので、具体的な判定基準は、適宜設定することが可能である。例えば、下記実施例では、血中LDL-TG値に基づき被験者をほぼ４等分し、各群における、15.6年以内の発症率を調査した結果、LDL-TG値が最も低い第１四分位における発症率を基準とすると、第２四分位、第３四分位及び第４四分位の各群における発症率は、高い統計学的有意差をもって統計学的に有意に増大していた。この際の第２四分位の下限値が17.1mg/dLであるので、例えば、閾値を17mg/dLに設定し、17mg/dLを超える場合に高リスクと判定することができる。あるいは、より高いリスクを高リスクとする場合には、例えば、中間値である22.7mg/dL以上を高リスクと判定することもできる。このように、閾値は、検出したいリスクの高さとの兼ね合いで任意に設定することが可能である。例えば、17mg/dL〜40mg/dLの範囲の任意の値、とりわけ25mg/dL〜35mg/dLの範囲の任意の値を閾値として設定することができる。なお、LDL-TG値は、民族等の集団により異なる可能性があるので、その集団内における中間値や平均値、第１四分位と第２四分位の境界の値等を閾値としたり、これらの近傍の値（例えば±20%以内の任意の値等）を閾値として判断することができる。さらに、適宜設定された閾値に比べて高ければ高いほどリスクが高くなるので、具体的な測定値に基づきリスクを判定することもできるし、測定値とイベントが発生する頻度を関連付けることにより、確率を数値で示すことも可能である。 In healthy individuals who have not developed cardiovascular disease, coronary heart disease or stroke, the higher the blood LDL-TG level, the higher the possibility of developing these diseases in the future. Therefore, it is possible to assist in the determination of the risk of developing these diseases in the future based on the measured blood LDL-TG value. The higher the blood LDL-TG level, the higher the possibility of developing these diseases in the future. Therefore, specific criteria can be set as appropriate. For example, in the following examples, subjects were divided into approximately 4 equal parts based on the blood LDL-TG level, and the incidence rate within 15.6 years in each group was investigated. As a result, the first quartile with the lowest LDL-TG level was obtained. Based on the incidence in Japan, the incidence in each of the second, third, and fourth quartiles was statistically significantly increased with high statistical significance. . Since the lower limit of the second quartile at this time is 17.1 mg / dL, for example, when the threshold is set to 17 mg / dL and exceeds 17 mg / dL, it can be determined that the risk is high. Alternatively, when a higher risk is set as a high risk, for example, an intermediate value of 22.7 mg / dL or more can be determined as a high risk. As described above, the threshold value can be arbitrarily set in consideration of the risk level to be detected. For example, an arbitrary value in the range of 17 mg / dL to 40 mg / dL, particularly an arbitrary value in the range of 25 mg / dL to 35 mg / dL can be set as the threshold value. Since the LDL-TG value may vary depending on the group of ethnic groups, etc., the intermediate value or average value in the group, the value of the boundary between the first quartile and the second quartile, etc. may be used as a threshold value. These neighboring values (for example, arbitrary values within ± 20%) can be determined as threshold values. Furthermore, the higher the threshold value is set, the higher the risk, so the risk can be determined based on the specific measurement value, and the probability can be determined by associating the measurement value with the frequency of occurrence of the event. Can also be indicated numerically.
同様に、血中LDL-TG値に基づき、発症中の脳卒中の診断を補助することができ、さらにStrokeが重症か否かを評価・判定することができる。例えばLDL-TGの定量値が20〜40mg/dL、好ましくは30mg/dLを超える場合、将来CVDまたはCHDまたはStrokeを発症するリスクが高くなると判断される。ただし、FCHL診断の基準値については民族等により異なることが考えられるためこの限りではない。また、CHDまたはStrokeを発症するリスクが高いと診断されたヒトの中でもLDL-TG値が40mg/dL、50mg/dLと高くなるに従い、その発症リスクは高まると判定し得る。 Similarly, based on the blood LDL-TG value, it is possible to assist in the diagnosis of stroke during onset, and it is possible to evaluate and determine whether Stroke is severe. For example, when the quantitative value of LDL-TG exceeds 20 to 40 mg / dL, preferably 30 mg / dL, it is determined that the risk of developing CVD, CHD or Stroke in the future increases. However, the reference value for FCHL diagnosis is not limited to this because it may differ depending on the ethnic group. Moreover, it can be determined that the onset risk increases as the LDL-TG value increases to 40 mg / dL and 50 mg / dL among humans diagnosed as having a high risk of developing CHD or Stroke.
以下、本発明を実施例に基づき具体的に説明する。もっとも、本発明は下記実施例に限定されるものではない。 Hereinafter, the present invention will be specifically described based on examples. However, the present invention is not limited to the following examples.
実施例１
米国におけるコホート試験の保存検体（血清）を用いて、LDL-TG濃度を測定した。具体的には、ベースラインの採血時にCHDまたは脳卒中を発症していない9,334例を対象として、LDL-TG濃度を測定した。そして、コホート研究の被験者を15.6年間追跡した結果より、その間のCHD、脳卒中発症（およびこれらを合わせたCVD）を調査し、測定項目との関連性を検証した。
Example 1
LDL-TG concentration was measured using a stored sample (serum) of a cohort test in the United States. Specifically, LDL-TG concentrations were measured in 9,334 patients who did not develop CHD or stroke at the time of baseline blood collection. Based on the results of follow-up of subjects in the cohort study for 15.6 years, we investigated CHD and stroke onset (and combined CVD) during that period, and verified the relationship with the measurement items.
LDL-TGの測定は、LDL-TG“Seiken”（デンカ生研株式会社社製）を使用した。 For the measurement of LDL-TG, LDL-TG “Seiken” (manufactured by Denka Seken Co., Ltd.) was used.
LDL-TG値を四分位に分け、COX proportional regression model モデルを用いて第1四分位群に対するハザード比を比較した。 The LDL-TG values were divided into quartiles, and the hazard ratio for the first quartile group was compared using the COX proportional regression model model.
結果を表１に示す。 The results are shown in Table 1.
表１に示すように、LDL-TG値が第1四分位に対し第2四分位、第3四分位、第4四分位ではCHD、脳卒中、CVDのハザード比が上昇し、リスクの増加が認められた。以上の結果はLDL-TG値を求めることにより将来的なCHD、脳卒中（およびこれらを合わせたCVD）発症のリスクの程度を判断できることを示すものである。 As shown in Table 1, the LDL-TG value is higher than the first quartile in the second quartile, third quartile, and fourth quartile, and the hazard ratio of CHD, stroke, and CVD is increased and the risk is increased. Increased. The above results indicate that the degree of risk of developing future CHD, stroke (and combined CVD) can be determined by determining the LDL-TG value.
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| JP2020030148A (en) * | 2018-08-23 | 2020-02-27 | デンカ生研株式会社 | Method for assisting detection of nonalcoholic steatohepatitis |
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| CN113539365A (en) * | 2020-04-16 | 2021-10-22 | 南京腾辰生物科技有限公司 | Methylation marker for early diagnosis of cardiovascular and cerebrovascular diseases |
| CN113539365B (en) * | 2020-04-16 | 2024-03-01 | 腾辰生物科技(上海)有限公司 | Methylation markers for early diagnosis of cardiovascular and cerebrovascular diseases |
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