JP6062439B2 - Evaluation markers for early kidney injury and methods for measuring them - Google Patents
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Description
本発明は、糖尿病に起因する腎機能疾患（特に糖尿病性腎症）の初期症状を的確に評価するための評価マーカーとその測定方法に関するものである。 The present invention relates to an evaluation marker for accurately evaluating an initial symptom of a renal function disease (particularly diabetic nephropathy) caused by diabetes and a method for measuring the same.
糖尿病は、持続した高血糖による血管合併症を招き、細小血管症（網膜症、腎症、神経障害）と大血管症（脳心血管系疾患）を合併する。この中でも細小血管症に区分される糖尿病性腎症は、１９９８年に慢性糸球体腎炎を抜いて日本の透析療法導入疾患の第１位となり、２００９年には、全透析療法導入者数のなかで４４．５％（１万６５４９人）を占め、医学的・医療経済的に大きな問題となっている。さらに、透析療法導入後の５年生存率は約５０％であり、今なお生命予後不良である。そのため、糖尿病性腎症の発症を早期に発見し、早期に治療する必要があるが、糖尿病性腎症は末期になるまで臨床症状に乏しく、積極的に検査を行わない場合見逃されやすい。したがって、糖尿病発症の早期から積極的に検査を行い、発症の診断や病期の評価を続けることが不可欠である。
糖尿病性腎症の診断には、腎生検による組織学的診断が必要である。しかし、すべての糖尿病患者に腎生検を施行することは困難であり、一般的には、臨床経過や網膜症などの合併症の有無に加え、尿検査・腎機能検査所見などを総合的に判断して診断することが多い。そのため現在では、糖尿病性腎症の早期診断は、微量アルブミン尿の出現により行われている。
日本では２００５年の日本糖尿病学会と日本腎臓学会の糖尿病性腎症合同委員会において、「糖尿病腎症の早期診断基準」が改訂された。この基準によると、尿中アルブミン測定対象者は、通常の試験紙法で尿蛋白陰性か、１＋程度の陽性を示す糖尿病患者である。尿は、なるべく午前中の随時尿を測定し、免疫測定法で尿中アルブミン値を定量し（同時に尿中クレアチニン値（Ｃｒ）も測定する）、３回測定中２回以上の尿中アルブミン値が３０〜２９９ｍｇ／ｇＣｒであれば、微量アルブミン尿とし、糖尿病性早期腎症と診断する。微量アルブミン尿だけでは糖尿病性腎症以外の諸病態も否定できないため、尿中ＩＶ型コラーゲン値の上昇や、腎肥大などの糖尿病性腎病変の存在を示唆する所見が診断の参考になる。
一方、初期の慢性腎臓病（ＣＫＤ：ｃｈｒｏｎｉｃ ｋｉｄｎｅｙ ｄｉｓｅａｓｅ）は、一般に自覚症状に乏しく、尿異常から始まり、徐々に腎機能が低下して末期腎不全へ進行する。したがって、採尿による尿異常の発見は、ＣＫＤ早期検出の唯一の手がかりといえる。中でも蛋白尿（微量アルブミン尿を含む）は、ＣＫＤの進展と密接に関わっており、最も重要な所見と言える。
最近、糖尿病患者において、“ＧＦＲ ｄｅｃｌｉｎｅｒ”（糸球体濾過量：ＧＦＲ：Ｇｌｏｍｅｒｕｌａｒ Ｆｉｌｔｒａｔｉｏｎ Ｒａｔｅ） あるいは“ｅａｒｌｙ ｒｅｎａｌ ｆｕｎｃｔｉｏｎ ｄｅｃｌｉｎｅ”と呼ばれる進行性の早期腎機能低下が報告されている。これについての明確な定義はないが、持続的蛋白尿を合併しない糖尿病患者で、腎機能が正常範囲内（６０ｍＬ／ｍｉｎ／１．７３ｍ２以上）にもかかわらず進行性に腎機能が低下し、アルブミン尿の増減に関係なく腎不全期へ至る症例が確認されるようになった。アルブミン尿の増加を認めないＧＦＲ ｄｅｃｌｉｎｅｒの症例では治療の開始が遅れてしまい、腎不全期への進行を十分に阻止することができていない可能性が考えられる。そのため、定期的な腎機能の評価により、早期にＧＦＲ ｄｅｃｌｉｎｅｒを同定することができれば、アルブミン尿の程度に関係なく、腎機能がまだ正常な時期において腎保護のための治療を開始することが可能となり、その後の腎機能の低下を抑制あるいは改善させることができるのではないかと期待される。これらのことから、微量アルブミン尿のみで糖尿病性腎症の早期診断を行うには限界があり、ＧＦＲ ｄｅｃｌｉｎｅｒの予測因子としてより鋭敏な糖尿病性腎症の早期臨床的指標の開発が求められている。
しかしながら、アルブミン尿の程度に関係なく、腎機能がまだ正常な時期において腎保護のための治療を開始することが可能となるマーカーや測定方法については、まだ充分な検討がなされていない状況である。なお、血中の特定のタンパク質に着目し、糖尿病性疾患の進行度を検査することが行なわれている。例えば、血中のアディポネクチンを測定することにより、早期に糖尿病の進展度を判定する方法が報告されている（非特許文献１）。Diabetes causes vascular complications due to persistent hyperglycemia, and is associated with microangiopathy (retinopathy, nephropathy, neuropathy) and macrovascular disease (cerebral cardiovascular disease). Among them, diabetic nephropathy, which is classified as microangiopathy, overtook chronic glomerulonephritis in 1998 and became the first place in Japan for introduction of dialysis therapy. Accounting for 44.5% (16,549), which is a major medical and medical economic problem. Furthermore, the 5-year survival rate after introduction of dialysis therapy is about 50%, and the life prognosis is still poor. Therefore, it is necessary to detect the onset of diabetic nephropathy at an early stage and treat it early, but diabetic nephropathy has few clinical symptoms until the end stage and is easily overlooked when it is not actively tested. Therefore, it is indispensable to actively test from the early stage of the onset of diabetes to continue diagnosis of the onset and evaluation of the stage.
Diagnosis of diabetic nephropathy requires histological diagnosis by renal biopsy. However, it is difficult to perform renal biopsy in all diabetic patients. In general, in addition to the clinical course and the presence or absence of complications such as retinopathy, urinalysis and renal function test findings are comprehensive. Judgment is often made based on judgment. Therefore, at present, early diagnosis of diabetic nephropathy is performed by the appearance of microalbuminuria.
In Japan, the “early diagnostic criteria for diabetic nephropathy” was revised in 2005 by the Joint Committee on Diabetic Nephropathy of the Japanese Diabetes Society and the Japanese Nephrology Society. According to this standard, the urinary albumin measurement subject is a diabetic patient who is urinary protein negative or positive about 1+ by the usual test paper method. For urine, measure urine as often as possible in the morning and quantify urinary albumin level by immunoassay (simultaneously measure urinary creatinine level (Cr)). Is 30-299 mg / gCr, microalbuminuria is assumed and diabetic early nephropathy is diagnosed. Since various conditions other than diabetic nephropathy cannot be ruled out by microalbuminuria alone, findings suggesting the presence of diabetic renal lesions such as increased urinary type IV collagen levels and renal hypertrophy are helpful.
On the other hand, chronic chronic kidney disease (CKD) is generally poor in subjective symptoms, starts with abnormal urine, gradually decreases in renal function, and progresses to end-stage renal failure. Therefore, the discovery of abnormal urine by collecting urine can be said to be the only clue for early detection of CKD. Among them, proteinuria (including microalbuminuria) is closely related to the progress of CKD and can be said to be the most important finding.
Recently, in diabetic patients, a progressive early decrease in renal function called “GFR decliner” (glomerular filtration rate) or “early real function declin” has been reported. Although there is no clear definition of this, in diabetic patients who do not have persistent proteinuria, the renal function is progressively decreased even though the renal function is within the normal range (60 mL / min / 1.73 m2 or more). Regardless of the increase or decrease in albuminuria, cases that lead to renal failure have been confirmed. In the case of GFR decliner that does not show an increase in albuminuria, the start of treatment is delayed, and it is possible that the progression to the renal failure stage could not be sufficiently prevented. Therefore, if GFR decliner can be identified early by regular evaluation of renal function, it is possible to start treatment for renal protection at a time when renal function is still normal, regardless of the degree of albuminuria Therefore, it is expected that the subsequent decline in renal function can be suppressed or improved. For these reasons, there is a limit to early diagnosis of diabetic nephropathy using only microalbuminuria, and there is a demand for the development of more sensitive early clinical indicators of diabetic nephropathy as a predictor of GFR decliner. .
However, regardless of the level of albuminuria, there has not yet been a sufficient study of markers and measurement methods that can start treatment for renal protection when renal function is still normal. . In addition, paying attention to a specific protein in the blood, examination of the progress of diabetic disease has been performed. For example, a method has been reported in which the progress of diabetes is determined at an early stage by measuring adiponectin in blood (Non-patent Document 1).
本発明の目的は、腎障害を早期に検出し、適切な生活指導を行い、腎障害を寛解・退縮させるための、腎障害リスクのマーカーとその測定評価方法を提供することにある。 An object of the present invention is to provide a renal injury risk marker and a method for measuring and evaluating the same for detecting renal injury early, providing appropriate life guidance, and ameliorating / reducing renal injury.
本発明者らは、非特許文献１の免疫複合体転移測定方法（ＩＣＴ−ＥＩＡ法）による高分子アディポネクチン測定方法を応用し、新たな腎変異の指標として尿中高分子アディポネクチンの可能性について検討して来た（ＰＣＴ／ＪＰ２０１２／０５５７３６）。その中で、本発明者らは更に検討を進め、具体的な患者のデータを評価検討した。その結果、患者の糸球体濾過能に違いにより、血中と尿中のアディポネクチン濃度に相違が見られた。即ち、糸球体濾過能が悪くなれば、尿中アディポネクチン濃度が高くなることから、血中のアディポネクチン濃度に対して尿中に濾出されるアディポネクチン比を求めたところ、糸球体濾過能が悪い患者では、健常人と比較して数十倍も高い値を示すことが明らかとなった。このことから、本発明者らは、尿中／血中アディポネクチン比が、アルブミン尿の傾向とは異なった、新たな腎障害の早期指標になり得ることを見出した。
更に、本発明者らは、健常者の尿中アディポネクチン値には、単分子アディポネクチンの占める割合が高いことから、単分子アディポネクチンを測定しないアディポネクチン測定方法により、尿中／血中アディポネクチン比の差が、より明白となることを見出した。即ち、ＨＭＷアディポネクチンをより特異的に測定し、尿中／血中アディポネクチン比を測定評価することにより、この尿中／血中アディポネクチン比をマーカーとして、腎障害の早期指標として使用できることを見出した。本発明者は、以上の知見に基づき、本発明を完成した。
即ち、本発明の要旨は以下の通りである。
（１）免疫複合体転移測定方法（ＩＣＴ−ＥＩＡ法）で測定された尿中のアディポネクチン濃度の測定値を血中のアディポネクチン濃度で除した数値であることを特徴とする、腎機能の評価マーカー。
（２）上記アディポネクチン濃度が、高分子量（ＨＭＷ）アディポネクチン濃度である、上記（１）に記載の腎機能の評価マーカー。
（３）上記（１）または（２）の数値に１０４を乗じた数値であることを特徴とする、上記（１）または（２）の腎機能の評価マーカー。
（４）上記（１）〜（３）に記載の評価マーカーを使用することを特徴とする、腎機能疾患の進行度評価方法。
（５）腎機能疾患が糖尿病性腎症であることを特徴とする、上記（４）に記載の腎機能疾患の進行度評価方法。
（６）上記（１）〜（３）の評価マーカーを用いて、腎機能疾患患者及び／又はその予備群に対して、介入療法を行うことを特徴とする、腎機能疾患の予防または治療方法。
（７）腎機能疾患が糖尿病性腎症であることを特徴とする、上記（６）に記載の腎機能疾患の予防または治療方法。
（８）介入療法が、食事療法、運動療法または薬物療法である、上記（６）または（７）に記載の糖尿病性疾患の予防または治療方法。The present inventors examined the possibility of high-molecular-weight adiponectin in urine as a new indicator of renal mutation by applying the high-molecular-weight adiponectin measuring method by the immune complex metastasis measuring method (ICT-EIA method) of Non-Patent
Furthermore, since the ratio of monomolecular adiponectin is high in the urinary adiponectin level of healthy individuals, the present inventors have found that the difference in the urine / blood adiponectin ratio is determined by the adiponectin measurement method that does not measure monomolecular adiponectin. And found that it becomes more obvious. That is, it was found that by measuring HMW adiponectin more specifically and measuring and evaluating the urine / blood adiponectin ratio, this urine / blood adiponectin ratio can be used as a marker as an early indicator of renal damage. The present inventor completed the present invention based on the above findings.
That is, the gist of the present invention is as follows.
(1) A renal function evaluation marker characterized by being a numerical value obtained by dividing a measured value of urine adiponectin concentration measured by an immune complex metastasis measuring method (ICT-EIA method) by adiponectin concentration in blood .
(2) The evaluation marker for renal function according to (1), wherein the adiponectin concentration is a high molecular weight (HMW) adiponectin concentration.
(3) The evaluation marker for renal function according to (1) or (2) above, which is a numerical value obtained by multiplying the numerical value of (1) or (2) by 10 4 .
(4) A method for evaluating the degree of progression of renal function disease, comprising using the evaluation marker according to (1) to (3) above.
(5) The method for evaluating the degree of progression of renal function disease according to (4) above, wherein the renal function disease is diabetic nephropathy.
(6) A method for preventing or treating renal function disease, comprising performing interventional therapy on a renal function disease patient and / or a reserve group thereof using the evaluation markers of (1) to (3) above .
(7) The method for preventing or treating a renal function disease according to the above (6), wherein the renal function disease is diabetic nephropathy.
(8) The method for preventing or treating diabetic disease according to (6) or (7) above, wherein the intervention therapy is diet therapy, exercise therapy or drug therapy.
本発明のＩＣＴ−ＥＩＡ法で得られた尿中と血中のアディポネクチン濃度の比を取り、これを腎機能の評価マーカーとすることにより、アルブミン尿の程度に関係なく、腎機能がまだ正常な時期において腎保護のための治療を開始することが可能となった。その結果、本発明の腎機能の評価マーカーの評価方法を用いることにより、糖尿病性疾患の診断として、特に問題の多い糖尿病性腎症の早期発見や、糖尿病予備群の腎機能障害の判定や介入療法後の効果判定にも有用である。 By taking the ratio of adiponectin concentration in urine and blood obtained by the ICT-EIA method of the present invention and using this as a marker for evaluating renal function, renal function is still normal regardless of the degree of albuminuria It became possible to start treatment for renal protection at certain times. As a result, by using the evaluation method of the renal function evaluation marker of the present invention, as a diagnosis of diabetic disease, the early detection of particularly problematic diabetic nephropathy, the determination or intervention of renal dysfunction in the pre-diabetes group It is also useful for determining effects after therapy.
図１は、本発明で使用した免疫複合体転移酵素免疫測定法（ＩＣＴ−ＥＩＡ法）の概略を表した図である。
図２は、被験者Ａ〜Ｄの腎機能に関して、ＣＫＤステージ分類を行なった図である。被験者ＡとＢは健常人であり、被験者ＣとＤでは腎機能の低下が予想されている。
図３は、被験者Ａ〜Ｄの血清サンプルと尿サンプルを用いて、Ｓｕｐｅｒｄｅｘカラムにより分子量別に分画し、血中及び尿中アディポネクチン分子存在様式を検討した図である。●：血中のアディポネクチン濃度（左軸）、◇：透析尿中のアディポネクチン濃度（右軸）を表わしている。フラクションＮｏ．４６前後のピークは、ＨＭＷアディポネクチンであり、フラクションＮｏ．５１前後のピークは、ＭＭＷアディポネクチンであり、フラクションＮｏ．５８前後のピークは、ＬＭＷアディポネクチンである。また、低分子領域のフラクションＮｏ．７８前後のピークは、単分子アディポネクチンに該当する。
図４は、被験者５０名の尿中アディポネクチン濃度（ｎｇ／ｍｇＣｒ）と腎機能指標のｅＧＦＲ（ｍＬ／ｍｉｎ／１．７３ｍ２）を測定して、その相関関係の有無を評価した図である。尿中のアディポネクチン濃度は、ｅＧＦＲと負の相関関係を示すことが明らかとなった。
図５は、図４で得られた評価データを集約し、ｅＧＦＲの各ステージ（Ｇ１、Ｇ２、Ｇ３ａ）に整理して、各ステージの中での尿中アディポネクチン濃度の平均とバラツキを表わした図である。尿中アディポネクチン濃度は、ステージＧ２においてステージＧ１よりも有意に高くなった。
図６は、図５と同様に図４で得られた評価データを集約し、ｅＧＦＲの各ステージ（Ｇ１、Ｇ２、Ｇ３ａ）に整理して、各ステージの中での尿中アルブミン濃度の平均とバラツキを表わした図である。尿中アルブミン濃度は、ステージＧ３ａにおいてステージＧ２よりも有意に高くなった。
図７は、上記のｅＧＦＲのステージＧ１における、尿中アディポネクチン濃度と尿中アルブミン濃度の各平均を１として、ステージＧ２とＧ３ａの各被験者の尿中アディポネクチン濃度と尿中アルブミン濃度の相対値を算定した。その数値をｅＧＦＲの各ステージ毎に表わした図である。FIG. 1 is a diagram showing an outline of an immunocomplex transferase immunoassay method (ICT-EIA method) used in the present invention.
FIG. 2 is a diagram in which CKD stage classification is performed on the renal functions of subjects A to D. FIG. Subjects A and B are healthy people, and subjects C and D are expected to have decreased renal function.
FIG. 3 is a diagram in which the serum and urine samples of subjects A to D were used for fractionation by molecular weight using a Superdex column, and the presence of adiponectin molecules in blood and urine was examined. ●: Adiponectin concentration in blood (left axis), ◇: Adiponectin concentration in dialyzed urine (right axis). Fraction No. The peak around 46 is HMW adiponectin. The peak around 51 is MMW adiponectin. The peak around 58 is LMW adiponectin. In addition, the low molecular weight fraction No. The peak around 78 corresponds to unimolecular adiponectin.
FIG. 4 is a diagram in which the urinary adiponectin concentration (ng / mgCr) and renal function index eGFR (mL / min / 1.73 m 2 ) of 50 subjects were measured, and the presence or absence of the correlation was evaluated. It was revealed that the adiponectin concentration in urine shows a negative correlation with eGFR.
FIG. 5 summarizes the evaluation data obtained in FIG. 4 and organizes it into each stage (G1, G2, G3a) of eGFR, and shows the average and variation of the urinary adiponectin concentration in each stage. It is. The urine adiponectin concentration was significantly higher in stage G2 than in stage G1.
FIG. 6 summarizes the evaluation data obtained in FIG. 4 in the same manner as in FIG. 5 and organizes the data into each stage (G1, G2, G3a) of eGFR, and the average urinary albumin concentration in each stage. It is a figure showing variation. The urinary albumin concentration was significantly higher in stage G3a than in stage G2.
FIG. 7 calculates the relative values of the urinary adiponectin concentration and the urinary albumin concentration of each subject of stage G2 and G3a, with the average of the urinary adiponectin concentration and the urinary albumin concentration at stage G1 of the above eGFR as 1. did. It is the figure which represented the numerical value for every stage of eGFR.
−本発明の第一の態様−
本発明の第一の態様は、免疫複合体転移測定方法（ＩＣＴ−ＥＩＡ法）で評価された血中と尿中のアディポネクチン濃度の数値を用いて、その比（尿中のアディポネクチン濃度／血中のアディポネクチン濃度）を取ることを特徴とする、腎機能の評価マーカーに関する発明である。
本発明の「免疫複合体転移酵素免疫測定方法（ＩＣＴ−ＥＩＡ法）」とは、非競合法（サンドイッチ）エンザイムイムノアッセイ法（ＥＩＡ）の高感度化した改良方法に関するものである（非特許文献１参照）。図１に示すように、ＩＣＴ−ＥＩＡ法では、使用する抗体の非特異吸着（バックグランド）を下げることができたので、多くの高分子生理活性物質のａｍｏｌレベル以下（ｚｍｏｌ）の測定が可能となっている（Ｈａｓｈｉｄａ Ｓ，ｅｔ ａｌ．，Ｂｉｏｔｅｃｈｎｏｌｏｇｙ Ａｎｎｕａｌ Ｒｅｖｉｅｗ Ｖｏｌ．１，（１９９５）ｐｐ４０３−４５１，Ｅｌｓｅｖｉｅｒ Ｓｃｉｅｎｃｅ Ｐｕｂｌｉｓｈｅｒｓ Ｂ．Ｖ．，Ａｍｓｔｅｒｄａｍ参照）。
本発明の「アディポネクチン」とは、脂肪細胞から分泌されるアディポサイトカインの１つで、２４４個のアミノ酸からなる蛋白質であり、血中には３量体（ＬＭＷ：Ｌｏｗ ｍｏｌｅｃｕｌａｒ ｗｅｉｇｈｔ）を基本構造として、６量体（ＭＭＷ：Ｍｉｄｄｌｅ ｍｏｌｅｃｕｌａｒ ｗｅｉｇｈｔ）、１８量体（ＨＭＷ：Ｈｉｇｈ ｍｏｌｅｃｕｌａｒ ｗｅｉｇｈｔ）など数種の多量体が存在することが報告されている。その血中濃度は、５〜１０μｇ／ｍＬという高濃度で存在しており、抗動脈硬化作用やインスリン抵抗性改善作用などの生理活性を有することが報告されている。さらに近年、これらの生理作用と多量体構造の役割が注目されており、高分子多量体レベルあるいは高分子多量体／総量比が、内臓脂肪型肥満や糖尿病、耐糖能、インスリン抵抗性、冠状動脈疾患、メタボリックシンドロームなどの様々な病態、さらに食事療法や手術による減量効果などにおいて、アディポネクチン総量よりもより相関が強いことも示され、アディポネクチンの分子別定量の重要性が示されている。さらに、肥満によるインスリン抵抗性においては、高分子量のアディポネクチンが特に関与することも見い出されている。また、アディポネクチンと腎障害について検討された報告によると、アディポネクチン欠損マウスでは、腎臓のポドサイトの足突起の融合が見られ、アルブミン尿が認められた。そこにアディポネクチンを補充すると、腎臓の病理組織の正常化及びアルブミン尿の改善が見られたという報告があることからも、アディポネクチンには腎障害に対して保護的な作用を有する可能性があることが示唆されている（Ｓｈａｒｍａ Ｋ ｅｔ ａｌ：Ａｄｉｐｏｎｅｃｔｉｎ ｒｅｇｕｌａｔｅｓ ａｌｂｕｍｉｎｕｒｉａ ａｎｄ ｐｏｄｏｃｙｔｅ ｆｕｎｃｔｉｏｎ ｉｎ ｍｉｃｅ．Ｊ Ｃｌｉｎ Ｉｎｖｅｓｔ１１８（５）：１６４５−１６５６，２００８．）。
なお、ＩＣＴ−ＥＩＡ法では、主にＨＭＷアディポネクチンを測定し、一部ＭＭＷアディポネクチンと単分子を測定していることが分かった。しかし、大変弱い反応であるが、ＬＭＷアディポネクチンも測定していることが分かった。
本発明の「腎機能」とは、腎臓の糸球体の濾過および再吸収機能のことを言う。
本発明の「評価マーカー」とは、尿中のアディポネクチン濃度を血中のアディポネクチン濃度で割った数値であり、例えば健常人の場合には、被験者ＡとＢに見られるように、０．３×１０−４以下である。一方、早期の腎機能障害（微量アルブミン尿）が示唆される被験者Ｄでは、４．０×１０−４となっている。このように、本発明の評価マーカーは、１０倍以上の数値の相違となっており、区別し易い指標である。更に、この数値に、１０４を乗じて、整数として、評価を行い易くすることもできる。
−本発明の第二の態様−
本発明の第二の態様は、上記の評価マーカーを使用し、腎機能疾患の進行度評価方法を行なう方法であり、更には、腎機能障害を有する患者に対して、介入療法を行うことを特徴とする、腎機能疾患の予防または治療方法に関する発明である。
本発明の「進行度評価方法」とは、尿中のアディポネクチン濃度／血中のアディポネクチンの比の変動を測定することにより、被験者の腎機能疾患の進行状況を的確に評価する方法である。この評価方法により、被験者の腎機能疾患の進行を抑制し改善するための治療剤の投与や生活指導等の介入療法が的確に行なえるようになる。
本発明の「介入療法」とは、プロトコールに基づく生活習慣（食事・身体活動中心）の改善または公知の高脂血症治療剤、糖尿病疾患治療剤の服用を言う。即ち、食事療法や運動療法を生活習慣の中に取り入れ、必要カロリーと消費カロリーのコントロールを行なうものであり、精神的なストレスを軽減するよう指導して、糖尿病性腎疾患の発症にかかわる要因を削除軽減することを行なう。更には、初期症状の腎機能疾患患者に対しては、本発明の評価マーカーの数値を下げるための薬物療法を行なうことを介入療法として含むものである。上記の腎機能疾患の進行度評価方法を用いて、早期の介入療法を開始することにより、例えば糖尿病性腎症などの重症度の腎機能障害への移行を抑制、回避し、症状を改善し、健康な状態に戻すことを言う。
なお、本発明の第二の態様で使用される用語で、第一の態様と共通する用語は同じ意味を表わすものである。そのため、特に言及することをしていない。-First embodiment of the present invention-
The first aspect of the present invention uses a numerical value of adiponectin concentration in blood and urine evaluated by an immune complex metastasis measurement method (ICT-EIA method), and the ratio (adiponectin concentration in urine / blood It is an invention relating to a marker for evaluating renal function.
The “immunoconjugate transferase immunoassay method (ICT-EIA method)” of the present invention relates to an improved method for improving the sensitivity of the non-competitive method (sandwich) enzyme immunoassay method (EIA) (Non-patent Document 1). reference). As shown in FIG. 1, in the ICT-EIA method, the nonspecific adsorption (background) of the antibody to be used could be lowered, so that it is possible to measure below amol level (zmol) of many macromolecular bioactive substances. (See, Hashida S, et al., Biotechnology Annual Review Vol. 1, (1995) pp 403-451, Elsevier Science Publishers BV, Amsterdam).
The “adiponectin” of the present invention is one of adipocytokines secreted from adipocytes and is a protein consisting of 244 amino acids, and a trimer (LMW: Low molecular weight) is used as a basic structure in the blood. It has been reported that several types of multimers such as hexamer (MMW: Middle molecular weight) and 18-mer (HMW: High molecular weight) exist. Its blood concentration exists at a high concentration of 5 to 10 μg / mL, and it has been reported that it has physiological activities such as an anti-arteriosclerosis effect and an insulin resistance improving effect. In recent years, these physiological actions and the role of multimeric structures have attracted attention, and the level of polymer multimer or polymer multimer / total ratio is determined by visceral fat type obesity, diabetes, glucose tolerance, insulin resistance, coronary artery It is also shown that there is a stronger correlation than the total amount of adiponectin in various diseases such as diseases, metabolic syndrome, etc., as well as the effect of weight loss due to diet and surgery, indicating the importance of quantification of adiponectin by molecule. Furthermore, it has also been found that high molecular weight adiponectin is particularly involved in insulin resistance due to obesity. According to a report on adiponectin and kidney damage, adiponectin-deficient mice showed a fusion of kidney podocyte foot processes and albuminuria. There is a report that supplementation with adiponectin has shown normalization of the pathological tissue of the kidney and improvement of albuminuria. Therefore, adiponectin may have a protective action against kidney damage. (Sharma K et al: Adipolectin regulations albuminum and posito function function in mice. J Clin Invest 118 (5): 1645-1656, 2008.).
In the ICT-EIA method, it was found that HMW adiponectin was mainly measured and a part of MMW adiponectin and a single molecule were measured. However, although it was a very weak reaction, it was found that LMW adiponectin was also measured.
The “renal function” of the present invention refers to the filtration and reabsorption function of the glomeruli of the kidney.
The “evaluation marker” of the present invention is a numerical value obtained by dividing the adiponectin concentration in urine by the adiponectin concentration in blood. For example, in the case of a healthy person, as seen in subjects A and B, 0.3 × 10 −4 or less. On the other hand, in the subject D in whom early renal dysfunction (microalbuminuria) is suggested, it is 4.0 × 10 −4 . Thus, the evaluation marker of the present invention has a numerical difference of 10 times or more, and is an easily distinguishable index. Furthermore, this numerical value can be multiplied by 10 4 to make it easy to perform evaluation as an integer.
-Second aspect of the present invention-
A second aspect of the present invention is a method for performing a method for evaluating the degree of progression of renal function disease using the above-described evaluation marker, and further comprising performing interventional therapy on a patient having renal dysfunction. The invention relates to a method for preventing or treating a renal function disease.
The “progression evaluation method” of the present invention is a method for accurately evaluating the progress of a renal functional disease in a subject by measuring a change in the urine adiponectin concentration / blood adiponectin ratio. By this evaluation method, intervention therapy such as administration of therapeutic agents and lifestyle guidance for suppressing and improving the progression of renal function disease in the subject can be performed accurately.
The “intervention therapy” of the present invention means improvement of lifestyle habits (mainly on diet and physical activity) based on a protocol, or taking a known therapeutic agent for hyperlipidemia and therapeutic agent for diabetes. In other words, dietary therapy and exercise therapy are incorporated into lifestyle habits, and the necessary calories and calories consumed are controlled. Instructions are given to reduce mental stress and factors related to the onset of diabetic kidney disease Deletion is reduced. Furthermore, for patients with early-onset renal function disease, an interventional therapy includes a drug therapy for lowering the value of the evaluation marker of the present invention. By using the above-mentioned method for evaluating the degree of progression of renal function disease, early intervention therapy is started, for example, the transition to severe renal dysfunction such as diabetic nephropathy is suppressed and avoided, and the symptoms are improved. Say, to return to a healthy state.
Note that terms used in the second aspect of the present invention and common to the first aspect represent the same meaning. Therefore, no particular mention is made.
次に実施例を挙げて本発明を更に説明するが、本発明はこれらに限定されるものではない。
（実施例１）ＩＣＴ−ＥＩＡ法による血中と尿中のアディポネクチン濃度の測定
（１）試薬
ａ）抗原：
標準リコンビナント・ヒト・アディポネクチンはＢｉｏＶｅｎｄｏｒ Ｌａｂ Ｍｅｄ Ｉｎｃ社（Ｐａｌａｃｋｅｃｈｏ，Ｃｚｅｃｈ Ｒｅｐｕｂｌｉｃ）より購入した。
ｂ）抗体：
モノクローナル抗ヒト・アディポネクチン抗体（ＢＡＦ１０６５及びＭＡＢ１０６５１）はＲ＆Ｄ Ｓｙｓｔｅｍｓ Ｉｎｃ．（ＭＮ，ＵＳＡ）より購入した。
ｃ）緩衝液：
緩衝液Ａ：０．１Ｍ ＮａＣｌ，０．０１％ ＢＳＡ，１ｍＭ ＭｇＣｌ２及び０．１％ ＮａＮ３を含む０．０１Ｍリン酸ナトリウム緩衝液（ｐＨ７．０）、
緩衝液Ｂ：５ｍＭ ＥＤＴＡを含む０．１Ｍリン酸ナトリウム緩衝液（ｐＨ６．０）、
緩衝液Ｃ：０．１Ｍ ＮａＣｌ，０．１％ ＢＳＡ，１ｍＭ ＭｇＣｌ２及び０．１％ ＮａＮ３を含む０．０１Ｍリン酸ナトリウム緩衝液（ｐＨ７．０）、
緩衝液Ｄ：０．４Ｍ ＮａＣｌ，０．１％ ＢＳＡ，１ｍＭ ＭｇＣｌ２及び０．１％ ＮａＮ３を含む０．０５Ｍリン酸ナトリウム緩衝液（ｐＨ７．０）、
緩衝液Ｅ：０．１Ｍ ＮａＣｌを含む０．０１Ｍリン酸ナトリウム緩衝液（ｐＨ７．０）
ｄ）血液サンプル：
被験者の血清サンプルを、１２時間絶食後の早朝空腹時に採血し（ＮＩＰＲＯ，２２Ｇホルダー付，大阪）、室温３０分静置後、遠心分離機（日立微量高速遠心機ＣＦ１６ＲＸＩＩ，日立工機，東京）で３０００ｒｐｍ、１０分遠心し、血清を得た。血清は、マイナス３０℃で凍結保存した。
ｅ）尿サンプル：
被験者の早朝第一尿（１０ｍＬ）に１／５０容量の５％ＢＳＡおよび５％ＮａＮ３を加え、透析まで４℃で保存した。２４時間以内に採取した尿を透析した。
透析は、透析用セルロースチューブ（透析膜ＵＣ８−３２−２５、三光純薬、東京）を用い、緩衝液（０．１ＭＮａＣｌを含む０．０１Ｍリン酸ナトリウム緩衝液ｐＨ７．０）に対して１．５ｍｌの上記尿サンプルの透析を行なった。透析後、透析尿の重量を秤量し、希釈倍率による補正を行なった。透析尿は直ちにマイクロチューブ（Ｓａｆｅ−Ｌｏｃｋ Ｔｕｂｅｓ１．５ｍｌ）に分注し、−２０℃で凍結保存を行なった。尚、尿中クレアチニン濃度は市販キット（クレアチニン−テストワコー、和光純薬、大阪）により測定した。
（２）方法
測定方法は、非特許文献１に記載のＩＣＴ−ＥＩＡ法を準用した。即ち、市販ＥＬＩＳＡ Ｋｉｔに添付されている標準高分子ヒト・アディポネクチン標準液を緩衝液Ｄで希釈したものを１００μＬ、または尿および血清を緩衝液Ｄで４０，０００倍希釈したもの１００μＬを用意し、これに酵素標識抗体及び捕捉用標識抗体を緩衝液Ｄに溶解した混合液１００μＬを加え、４℃ １６時間インキュベーションし、酵素標識抗体・アディポネクチン・捕捉用標識抗体の３者からなる免疫複合体を形成させた。次いで、この反応液に抗ＤＮＰ−ＩｇＧ不溶化ポリスチレンビーズ１個を加え、０．５時間反応させビーズ上に免疫複合体を捕捉した。このビーズを緩衝液Ａ（２ｍＬ）で２回洗浄後、緩衝液Ｃに溶解した２ｍＭ ＤＮＰ−Ｌｙｓ（１５０μＬ）と０．５時間反応させ、ビーズから免疫複合体を溶出させた。抗ＤＮＰ−ＩｇＧ不溶化ポリスチレンビーズを除去した後、溶出液にストレプトアビジン不溶化ポリスチレンビーズ１個を加え、さらに０．５時間反応させ、第２のビーズ上に免疫複合体を転移させた。ビーズとの反応はすべて２５℃で２１０回／分の振盪下に行った。再びビーズを緩衝液Ｃ（２ｍＬ）で３回洗浄後、ビーズ上に転移されたβ−Ｄ−ｇａｌａｃｔｏｐｙｒａｎｏｓｉｄｅ（蛍光基質；４ＭＵＧ）（４００μＬ）を用いて３０℃インキュベーションし、０．１Ｍグリシンナトリウム緩衝液（ｐＨ １０．５）（２ｍＬ）を加え反応を停止後、蛍光分光光度計（Ｆ−２５００，日立）を用い測定した。なお、励起波長３６０ｎｍ蛍光波長４５０ｎｍを用い、蛍光強度は１０−８Ｍ ４ＭＵを１００として換算した（Ｈａｓｈｉｄａ Ｓ ａｎｄ Ｉｓｈｉｋａｗａ Ｅ ：Ｄｅｔｅｃｔｉｏｎ ｏｆ ｏｎｅ ｍｉｌｌｉａｔｔｏｍｏｌｅ ｏｆ ｆｅｒｒｉｔｉｎ ｂｙ ｎｏｖｅｌ ａｎｄ ｕｌｔｒａｓｅｎｓｉｔｉｖｅ ｅｎｚｙｍｅ ｉｍｍｕｎｏａｓｓａｙ．Ｊ Ｂｉｏｃｈｅｍ １０８：９６０−９６４．１９９０．）。
（３）測定結果
ａ）被験者：
腎機能指標としての尿中アディポネクチンを評価するために、異なる性、年齢、腎機能を有する対象者を選別し、血中及び尿中アディポネクチンの検討を行った。若い健常な腎機能を有する２０代女性Ａ、健常な腎機能を有する４０代女性Ｂ、年齢と共に腎機能推定指標であるｅＧＦＲ（ｍＬ／ｍｉｎ／１．７３ｍ２）が低下傾向を見せている５０代男性Ｃ、ｅＧＦＲは、Ｃと変わらないが、微量アルブミン尿傾向にあることから、腎糸球体の濾過能の低下が示唆される６０代肥満男性Ｄを対象者とした。
ｂ）評価結果：
まず、一般生化学検査の結果、血中クレアチニン濃度及び年齢と性からｅＧＦＲを算出した。その結果を表１に示す。被験者Ａは、９７．４ｍＬ／ｍｉｎ／１．７３ｍ２、被験者Ｂは、１０７．２ｍＬ／ｍｉｎ／１．７３ｍ２、被験者Ｃは、４８．２ｍＬ／ｍｉｎ／１．７３ｍ２、被験者Ｄは、５０．０ｍＬ／ｍｉｎ／１．７３ｍ２であった。
ＩＣＴ−ＥＩＡ法による血中アディポネクチン濃度は、被験者Ａにおいて、２１．３μｇ／ｍＬと高く、被験者ＢもＡと同様に２０．７μｇ／ｍＬと高い。しかし被験者Ｃでは、２．５８μｇ／ｍＬと４μｇ／ｍＬを下回っていることから、低アディポネクチン血症を示した。被験者Ｄは、１４．８μｇ／ｍＬと性別や年齢、基礎疾患があり、運動習慣がないにも関わらず高値を示していた。
血液と同様に測定した尿中アディポネクチン濃度は、被験者Ａにおいて、０．５６ｎｇ／ｍｇＣｒであり、被験者Ｂは、０．３０ｎｇ／ｍｇＣｒであった。また、被験者Ｃの尿中アディポネクチン濃度は、２．１６ｎｇ／ｍｇＣｒと被験者ＡやＢよりも高く、被験者Ｄも同様に５．９３ｎｇ／ｍｇＣｒと高値を示した。
次に血中のアディポネクチン濃度に対して尿中に濾出されるアディポネクチン濃度の比を求めたところ、被験者Ａは、０．３×１０−４、Ｂで０．１×１０−４に対し、被験者ＣとＤは、８．４×１０−４と４．０×１０−４であり、数十倍高い値を示した。このように、図２で示される被験者の腎機能のステージが、本発明の評価マーカー（尿中／血中アディポネクチン比）で明確に区別でき、被験者Ｄのように微量アルブミン尿の範囲に該当する対象者であっても、この評価マーカーを使用することで早期診断ができ、早期介入療法が開始できることになった。
（実施例２）被験者Ａ〜Ｄの血液及び尿におけるアディポネクチンの存在様式の検討
Ｓｕｐｅｒｄｅｘ２００を用いた分子ふるいカラムにより、希釈した被験者Ａ〜Ｄの血清及び尿を分子量別に分画し、その分画をＩＣＴ−ＥＩＡ法により測定を行った。その結果、血清中の各対象者のアディポネクチン存在様式は、これまでの血中様式と同様にＨＭＷアディポネクチンとＭＭＷアディポネクチンがほとんどであることが分かった。
ａ）尿中のアディポネクチンの存在様式：
尿中のアディポネクチンは、以下の表２に示すように、腎機能が正常な被験者Ａ、Ｂでは、絶対量としてはかなり少ないが、これまでと同様にＨＭＷ、ＭＭＷ、ＬＭＷ及び単分子アディポネクチンが検出された。
ｂ）血中のアディポネクチンの存在様式：
被験者Ａ〜Ｄの血清中のアディポネクチン存在様式は、図３（●）に示されるように、ＨＭＷアディポネクチンとＭＭＷアディポネクチンがほとんどであることが分かった。尿中のアディポネクチンの存在様式が顕著に異なることが示されている。従って、この尿中と血中のＨＭＷアディポネクチンの濃度を測定し、比（尿中アディポネクチン濃度／血中アディポネクチン濃度）を取れば、より顕著な指標として、腎機能障害の初期症状を評価できることになる。
（実施例３）腎機能の指標ｅＧＦＲに対する尿中のアディポネクチン値とアルブミン値の相関性の評価
ａ）ｅＧＦＲに対する相関性：
健康診断において被験者７０名から採取した尿中のアディポネクチン濃度（ｎｇ／ｍｇＣｒ）とアルブミン濃度（μｇ／ｍｇＣｒｅ）を実施例１に準じて測定した。また、同時に被験者の血中クレアチニン濃度及び年齢と性からｅＧＦＲを算出した。
その結果、尿中のアルブミン濃度とｅＧＦＲの間に有意な相関関係は認められなかったが、尿中のアディポネクチン濃度とｅＧＦＲの間には、図４に示されるように、有意な負の相関関係が認められた。なお、血中のアディポネクチン濃度（μｇ／ｍＬ）とｅＧＦＲの間には、有意な相関関係は見出せなかった。
ｂ）ｅＧＦＲの各ステージ（Ｇ１、Ｇ２、Ｇ３ａ）に対する相関性：
前項の被験者７０名のｅＧＦＲ値から、図２に示される各ステージに被験者を分類した。
ステージＧ１（９０以上）、ステージＧ２（６０〜８９）、ステージＧ３ａ（４５〜５９）に分類される対象者の尿中アディポネクチン濃度（ｎｇ／ｍｇＣｒ）と尿中アルブミン濃度（μｇ／ｍｇＣｒｅ）をまとめた。
その結果を図５と６に示した。上記各ステージ間における尿中アディポネクチン濃度および尿中アルブミン濃度を比較すると、尿中アディポネクチン濃度はＧ２期においてＧ１期より有意に高値を示した。一方、尿中アルブミン濃度はＧ３ａ期においてＧ２期より有意に高値を示した。
以上の結果は、尿中アディポネクチン濃度は尿中アルブミン濃度より腎障害予知マーカーとしての有用であることを示している。
ｃ）ｅＧＦＲの各ステージにおける尿中アディポネクチン濃度と尿中アルブミン濃度の相対値：
ｅＧＦＲのステージＧ１の被験者の尿中アディポネクチン濃度と尿中アルブミン濃度の中央値をそれぞれ１とする。次に、ステージＧ２とステージＧ３ａの被験者の尿中アディポネクチン濃度と尿中アルブミン濃度がそれぞれ何倍になっているかを算定した。
その結果を図７に示した。即ち、ステージＧ２において、尿中アディポネクチン相対値が尿中アルブミン相対値に比べ、有意に高値を示した。
以上の結果は、腎障害予知マーカーとして、尿中アディポネクチン相対値の有用性を示している。EXAMPLES Next, although an Example is given and this invention is further demonstrated, this invention is not limited to these.
(Example 1) Measurement of adiponectin concentration in blood and urine by ICT-EIA method (1) Reagent a) Antigen :
Standard recombinant human adiponectin was purchased from BioVendor Lab Med Inc (Parackecho, Czech Republic).
b) Antibody :
Monoclonal anti-human adiponectin antibodies (BAF1065 and MAB10651) are available from R & D Systems Inc. (MN, USA).
c) Buffer:
Buffer A: 0.01 M sodium phosphate buffer (pH 7.0) containing 0.1 M NaCl, 0.01% BSA, 1 mM MgCl 2 and 0.1% NaN 3
Buffer B: 0.1 M sodium phosphate buffer (pH 6.0) containing 5 mM EDTA,
Buffer C: 0.01 M sodium phosphate buffer (pH 7.0) containing 0.1 M NaCl, 0.1% BSA, 1 mM MgCl 2 and 0.1% NaN 3
Buffer D: 0.05 M sodium phosphate buffer (pH 7.0) containing 0.4 M NaCl, 0.1% BSA, 1 mM MgCl 2 and 0.1% NaN 3
Buffer E: 0.01 M sodium phosphate buffer (pH 7.0) containing 0.1 M NaCl
d) Blood sample :
Serum samples of subjects were collected on an early fasting after fasting for 12 hours (NIPRO, with 22G holder, Osaka), allowed to stand at room temperature for 30 minutes, and then centrifuged (Hitachi Micro High Speed Centrifuge CF16RXII, Hitachi Koki, Tokyo) And centrifuged at 3000 rpm for 10 minutes to obtain serum. Serum was stored frozen at minus 30 ° C.
e) Urine sample :
1/50 volumes of 5% BSA and 5% NaN 3 were added to the subject's early morning first urine (10 mL) and stored at 4 ° C. until dialysis. The urine collected within 24 hours was dialyzed.
For dialysis, a cellulose tube for dialysis (dialysis membrane UC8-32-25, Sanko Junyaku Co., Ltd., Tokyo) was used, and 1. against a buffer solution (0.01 M sodium phosphate buffer pH 7.0 containing 0.1 M NaCl). Dialysis of 5 ml of the urine sample was performed. After dialysis, the weight of dialysis urine was weighed and corrected by the dilution factor. The dialyzed urine was immediately dispensed into microtubes (Safe-Lock Tubes 1.5 ml) and stored frozen at -20 ° C. The urinary creatinine concentration was measured with a commercially available kit (creatinine-Test Wako, Wako Pure Chemicals, Osaka).
(2) Method The ICT-EIA method described in
(3) Measurement result a) Subject :
In order to evaluate urinary adiponectin as an indicator of renal function, subjects with different gender, age and renal function were selected and examined for blood and urine adiponectin. A woman in her 20s with healthy healthy kidney function, a woman in her 40s with healthy kidney function, and eGFR (mL / min / 1.73 m 2 ), which is a renal function estimation index, shows a decreasing trend with age 50 Teenage male C and eGFR were the same as C, but because they tend to have microalbuminuria, 60-year-old obese male D, whose reduction in renal glomerular filtration ability is suggested, was used as the subject.
b) Evaluation results :
First, as a result of a general biochemical test, eGFR was calculated from blood creatinine concentration, age and sex. The results are shown in Table 1. Subject A is 97.4 mL / min / 1.73 m 2 , Subject B is 107.2 mL / min / 1.73 m 2 , Subject C is 48.2 mL / min / 1.73 m 2 , Subject D is 50 It was 0.0 mL / min / 1.73 m 2 .
The blood adiponectin concentration by the ICT-EIA method is as high as 21.3 μg / mL in the subject A, and the subject B is as high as 20.7 μg / mL as in the case of A. However, subject C showed hypoadiponectinemia because it was below 2.58 μg / mL and 4 μg / mL. Subject D had 14.8 μg / mL, had sex, age, and underlying disease, and showed a high value despite having no exercise habits.
The urinary adiponectin concentration measured in the same manner as in blood was 0.56 ng / mgCr in subject A and 0.30 ng / mgCr in subject B. Further, the urinary adiponectin concentration of subject C was higher than those of subjects A and B, 2.16 ng / mgCr, and subject D similarly showed a high value of 5.93 ng / mgCr.
Next, when the ratio of the adiponectin concentration filtered out in the urine to the adiponectin concentration in the blood was determined, the subject A was 0.3 × 10 −4 and B was 0.1 × 10 −4 . C and D were 8.4 × 10 −4 and 4.0 × 10 −4 , which were several tens of times higher. As described above, the stage of the renal function of the subject shown in FIG. 2 can be clearly distinguished by the evaluation marker (urine / blood adiponectin ratio) of the present invention, and corresponds to the range of microalbuminuria as in the subject D. Even subjects were able to make early diagnosis and start early intervention therapy by using this evaluation marker.
(Example 2) Examination of the existence pattern of adiponectin in the blood and urine of subjects A to D The serum and urine of diluted subjects A to D were fractionated by molecular weight using a molecular sieve
a) Mode of existence of adiponectin in urine:
As shown in Table 2 below, urinary adiponectin was detected in HMW, MMW, LMW, and unimolecular adiponectin as before, although the absolute amount was considerably small in subjects A and B with normal renal function. It was done.
b) Presence of adiponectin in blood:
As shown in FIG. 3 (●), it was found that HMW adiponectin and MMW adiponectin are mostly present in the sera of subjects A to D. It has been shown that the presence of adiponectin in urine is significantly different. Therefore, if the concentration of HMW adiponectin in urine and blood is measured and the ratio (urine adiponectin concentration / blood adiponectin concentration) is taken, the initial symptoms of renal dysfunction can be evaluated as a more prominent indicator. .
(Example 3) Evaluation of correlation between urinary adiponectin level and albumin level with respect to kidney function index eGFR a) Correlation with eGFR:
The urine adiponectin concentration (ng / mgCr) and albumin concentration (μg / mgCre) collected from 70 subjects in the health check were measured according to Example 1. At the same time, eGFR was calculated from the blood creatinine concentration, age and sex of the subject.
As a result, no significant correlation was found between albumin concentration in urine and eGFR, but a significant negative correlation was observed between urine adiponectin concentration and eGFR, as shown in FIG. Was recognized. A significant correlation could not be found between the blood adiponectin concentration (μg / mL) and eGFR.
b) Correlation for each stage (G1, G2, G3a) of eGFR:
From the eGFR values of 70 subjects in the previous section, subjects were classified into each stage shown in FIG.
Summary of urinary adiponectin concentration (ng / mgCr) and urinary albumin concentration (μg / mgCre) of subjects classified into stage G1 (90 or more), stage G2 (60 to 89), and stage G3a (45 to 59) It was.
The results are shown in FIGS. Comparing the urinary adiponectin concentration and the urinary albumin concentration between the above stages, the urinary adiponectin concentration was significantly higher in the G2 phase than in the G1 phase. On the other hand, the urinary albumin concentration was significantly higher in the G3a phase than in the G2 phase.
The above results indicate that the urinary adiponectin concentration is more useful as a marker for predicting renal damage than the urinary albumin concentration.
c) Relative values of urinary adiponectin concentration and urinary albumin concentration at each stage of eGFR:
The median values of urinary adiponectin concentration and urinary albumin concentration of eGFR stage G1 subjects are set to 1, respectively. Next, it was calculated how many times the urinary adiponectin concentration and the urinary albumin concentration of the subjects of stage G2 and stage G3a were respectively increased.
The results are shown in FIG. That is, in stage G2, the urinary adiponectin relative value was significantly higher than the urinary albumin relative value.
The above results indicate the usefulness of the relative value of urinary adiponectin as a marker for predicting renal damage.
本発明の腎機能障害の評価マーカーとその評価方法は、採取された尿中に存在するアディポネクチン濃度および同時に採取された血液の中に存在するアディポネクチン濃度との比を評価マーカーの数値とするものであり、得られた評価マーカーの数値から、腎臓の初期機能障害の状況を早期に把握することが可能になった。また、本発明の評価方法を用いて、健康診断への応用と介入療法と組み合わせての医療システムの構築が可能となる。 The evaluation marker for renal dysfunction according to the present invention and the evaluation method use the ratio of the adiponectin concentration present in the collected urine and the adiponectin concentration present in the collected blood as the numerical value of the evaluation marker. Yes, it became possible to grasp early the state of the initial dysfunction of the kidney from the numerical value of the obtained evaluation marker. In addition, using the evaluation method of the present invention, it is possible to construct a medical system in combination with application to health checkup and intervention therapy.
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