CN104737020B - The evaluation index of nephropathy and its assay method in early days - Google Patents
The evaluation index of nephropathy and its assay method in early days Download PDFInfo
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Abstract
The present invention is to provide the early diagnosis index of initial symptoms of renal function disease and its evaluation method as problem.Particularly, to provide the evaluation index of diagnosis of the initial symptoms that can definitely carry out nephrosis and its evaluation method as problem.The present inventor, in order to solve above-mentioned problem, uses immune complex transferase linked immunoassay method (ICT EIA method), is used for evaluation index by neutralize adiponectin concentration in blood with urine, finds more precisely to carry out the evaluation of renal function.As a result, become also can definitely grasp the handicapped effect carrying out situation or interventional therapy of kidney.
Description
[technical field]
The present invention relates to for definite renal function disease (the particularly glycosuria evaluated caused by diabetes
Characteristic of disease ephrosis) the evaluation index of initial symptoms and its assay method.
[background technology]
Diabetes cause, by the vascular complication caused by lasting hyperglycaemia, merging minute blood vessel disease
(nethike embrane disease, kidney disease, neurological disorder) and big blood vessel disease (brain disease of cardiovascular system).?
The most also the nephrosis of minute blood vessel disease it is divided at number chronic glomerulus in 1998
Ephritis is the 1st of the dialysis therapy importing disease of Japan, in 2009, at full dialysis therapy
Importing person's number accounts for 44.5% (10,000 6549 people), medical science-medical economics becomes big
Problem.Furthermore, 5 years survival rates after dialysis therapy imports are about 50%, currently still life
Prognosis mala.Therefore, have early detection nephrosis sends out disease, necessity of early treatment
Property, but nephrosis lacks clinical symptoms latter stage to becoming, easy when the most actively checking
Leak through.Thus, actively check from the early stage of diabetes complications, persistently send out disease diagnosis or
The evaluation of stadium is indispensable.
In the diagnosis of nephrosis, the histodiagnosis of renal biopsy it is necessary.
But, whole diabetics is implemented renal biopsy is difficult, it is however generally that, remove
Clinical through or nethike embrane disease etc. complication presence or absence outside, comprehensive descision uroscopy-kidney merit
The situation that can check finding etc. and diagnose is many.Therefore present, the early diagnosis of nephrosis
Carried out by the appearance of microalbuminuria.
In Japan, the Japan Diabetes association of 2005 and the diabetic keratopathy kidney of Japan's nephrology meeting
In sick Committee on Contracts, " the early diagnosis benchmark of diabetogenous nephrosis disease " is changed.According to this benchmark,
In urine, albumin measuring object is to show Urine proteins feminine gender or 1+ degree in common ph test paper ph method
Positive diabetic.Urine measures the urine at any time in the morning as far as possible, legal with immunoassays
Albumin value (simultaneously the most also measuring creatinine values (Cr) in urine) in amount urine, measure for 3 times in 2
In secondary above urine, albumin value is 30～299mg/gCr, then as microalbuminuria, examine
Break as diabetic keratopathy kidney disease in early days.In only microalbuminuria, owing to also cannot negating glycosuria
All morbid state beyond characteristic of disease ephrosis, in urine, IV Collagen Type VI value raises or points out hypernephrotrophy etc.
The finding of the existence of diabetic nephropathy becomes the reference of diagnosis.
On the other hand, the CKD (CKD:chronic kidney disease) at initial stage
Generally speaking lack subjective symptoms, start from urine is abnormal, lentamente renal function reduce and to latter stage
Renal failure is in progress.Thus, the discovery that the urine adopting urine is abnormal become CKD detection in early days
Unique means.The most also it is albuminuria (including microalbuminuria) and CKD
It is in progress closely related, the most most important finding.
Recently, report in diabetic and be referred to as " GFRdecliner " (glomerular filtration
Amount: GFR:Glomerular Filtration Rate) or " Renal function in early period reduce (early renal
Function decline) " progressive Renal function in early period reduce.Without the clear and definite definition to it,
But in the most persistently merging albuminuretic diabetic, although renal function is in normal range (NR)
(60mL/min/1.73m2Above), but the reduction of Progressive symmetric erythrokeratodermia renal function, with the increasing of albuminuria
Subtract irrelevant, confirm the case to renal failure stage.Confirming the increase less than albuminuria
GFRdecliner case in, it is contemplated that the starting for the treatment of is delayed by, it is impossible to fully ground resistance
Only to the possibility of the carrying out of renal failure stage.Therefore, by the evaluation of regularly renal function,
Can identify GFRdecliner in early days, then irrelevant with the degree of albuminuria, renal function still exists
Normal epoch starts to be possibly realized for the treatment of kidney protection, expects the kidney merit that whether can make thereafter
The reduction suppression of energy or improvement.By these, only carry out nephrosis by microalbuminuria
Early diagnosis have limitation, it is desirable to as the sharper sugar of the predictive factor of GFRdecliner
The exploitation of the index of the early clinic of urine characteristic of disease ephrosis.
But, irrelevant for the degree with albuminuria, renal function is still opened when normal epoch
Begin, for the index that is possibly realized for the treatment of of kidney protection or assay method, to be still in and do not carry out fully
The situation of discussion.Further, the specific protein being concerned about in blood, carry out inspection glycosuria
The degree of carrying out of characteristic of disease disease.Such as, report has by measuring the adiponectin in blood, carly fruit drop
The method (non-patent literature 1) of the progress degree of diabetes.
[prior art literature]
[non-patent literature]
Non-patent literature 1: bridge field is the most first-class, " exploitation of high sensitivity adiponectin determination method and to
The application of its motion-nutrition guide " Tokushima unity and coherence in writing university research summary No. 78 page 7～20
(2009)
[summary of the invention]
[inventing technical task to be solved]
It is an object of the invention to provide for detect in early days nephropathy, carry out suitable Life Guidance,
Make index and its evaluation of measuring method of the nephropathy risk that nephropathy alleviates-shrink back.
[solving the technical scheme of problem]
The immune complex transfer assay method of the present inventor's applications exploiting non-patent literature 1
The macromolecule adiponectin assay method of (ICT-EIA method), as the index pair of new kidney variation
In urine, the possibility of macromolecule adiponectin has carried out inquiring into (PCT/JP2012/055736).Its
In, the present inventor inquires into again, evaluates the data inquiring into concrete patient.As a result, by suffering from
The difference of the glomerular filtration ability of person, the adiponectin concentration in blood and in urine sees difference.
That is, glomerular filtration is less able, then in urine, adiponectin concentration uprises, so relative in blood
Adiponectin concentration obtain the adiponectin that leaches in urine than time, learn in glomerular filtration ability
Patient in, compare the value showing that decades of times is high with Healthy People.Therefore, the inventors discovered that in urine/
In blood, adiponectin referred to than the early stage that can become new nephropathy different from the tendency of albuminuria
Mark.
Furthermore, the inventors discovered that, unimolecule adiponectin in adiponectin value in the urine of healthy person
Shared ratio is high, so the adiponectin assay method by accident order molecule adiponectin is brighter
In albiduria/blood in the difference of adiponectin ratio.I.e. find, joined by more specific the HMW fat that measures
Element, evaluation of measuring urine in/blood in adiponectin ratio, in urinating with this/blood in adiponectin be used for index,
Can use as the early stage index of nephropathy.The present inventor completes the present invention based on above opinion.
That is, the main points of the present invention are as follows.
(1) evaluation index of renal function, it is characterised in that be to use immune complex transfer assay
The measured value of the adiponectin concentration in the urine that method (ICT-EIA method) measures is divided by the fat in blood
The numerical value of connection element concentration.
(2) evaluation index of the renal function described in above-mentioned (1), the most above-mentioned adiponectin is dense
Degree is HMW (HMW) adiponectin concentration.
(3) above-mentioned (1) or the evaluation index of renal function of (2), it is characterised in that on being
The numerical value stating (1) or (2) is multiplied by 104Numerical value.
(4) degree of the carrying out evaluation method of renal function disease, it is characterised in that use above-mentioned
(1) evaluation index～described in (3).
(5) degree of the carrying out evaluation method of the renal function disease described in above-mentioned (4), its feature
Being, renal function disease is nephrosis.
(6) prevention of renal function disease or methods for the treatment of, it is characterised in that use above-mentioned
(1)～the evaluation index of (3), for renal function disease patient and/or its suffer from danger person and enter
Row interventional therapy.
(7) prevention of the renal function disease described in above-mentioned (6) or methods for the treatment of, its feature
Being, renal function disease is nephrosis.
(8) above-mentioned (6) or (7) described in the prevention of diabetic condition or methods for the treatment of,
Wherein interventional therapy is have dinner therapy, kinesiatrics or medicinal treatment.
[invention effect]
The obtained adiponectin concentration urinated in neutralization blood is taken by the ICT-EIA method of the present invention
Ratio, by the evaluation index as renal function, thus irrelevant with the degree of albuminuria,
Renal function still starts when normal epoch to become possibility for the treatment of kidney protection.As a result, pass through
Use the evaluation method of the evaluation index of the renal function of the present invention, examining as diabetic condition
Disconnected, danger is suffered from early detection or diabetes for the many nephrosis of particularly problem
Effect after the judgement of the renal dysfunction of person or interventional therapy judges the most useful.
[accompanying drawing explanation]
Fig. 1 is to represent the immune complex transfer enzymoimmunoassay used in the present invention
The figure of the outline of (ICT-EIA method).
Fig. 2 is the figure that the renal function about experimenter A～D carries out CKD Stage Classification.It is subject to
Examination person A and B is Healthy People, the reduction of anticipated renal function in experimenter C and D.
Fig. 3 is blood serum sample and the urine samples using experimenter A～D, Superdex post depend on
Molecular weight carries out classification separation, and in inquiring in blood and urinating, adiponectin molecule exists the figure of pattern.●:
Represent the adiponectin concentration (left axle) in blood, ◇: represent that the adiponectin concentration in dialysis urine is (right
Axle).Peak before and after fraction No.46 is HMW adiponectin, and the peak before and after fraction No.51 is
MMW adiponectin, the peak before and after fraction No.58 is LMW adiponectin.It addition, low molecule
Peak before and after fraction No.78 in region corresponds to unimolecule adiponectin.
Fig. 4 is adiponectin concentration (ng/mgCr) and renal function in the urine measuring experimenter 50
EGFR (the mL/min/1.73m of index2), evaluate and scheme with presence or absence of its dependency relation.Display
Adiponectin concentration in urine becomes, with eGFR, the dependency relation born.
Fig. 5 is to represent, the evaluating data obtained in intensive Fig. 4, arranges each rank into eGFR
Section (G1, G2, G3a), in the urine among each stage, the mean value of adiponectin concentration is with inclined
The figure of difference.In urine, adiponectin concentration is compared stage G1 at stage G2 and is significantly more uprised.
Fig. 6 is to represent, the evaluating data obtained in intensive Fig. 4 in the same manner as Fig. 5, arranges and is
Each stage (G1, G2, G3a) of eGFR, albumin concentration in the urine among each stage
The average and figure of deviation.In urine, to compare stage G2 at stage G3a more notable for albumin concentration
Uprise.
Fig. 7 is with white in adiponectin concentration in the urine in the stage G1 of above-mentioned eGFR and urine
Protein concentration each average as 1, calculates fat in the urine of each experimenter determining stage G2 and G3a
The relative value of albumin concentration in connection element concentration and urine, by this numerical value in every each stage of eGFR
The figure represented.
[embodiment]
[embodiment of the first of the present invention]
The embodiment of the first of the present invention relates to the invention of the evaluation index of renal function, and it is special
Levy and be, in the blood that use immune complex transfer assay method (ICT-EIA method) is evaluated
With the numerical value of the adiponectin concentration in urine, take it than (the fat connection in the adiponectin concentration/blood in urine
Element concentration).
" immune complex transferase linked immunoassay method (ICT-EIA method) " of the present invention is
The changing of high-sensitivity about non-competing combined techniques (sandwich) enzyme immunoassay (EIA)
The method (with reference to non-patent literature 1) of good recipe method.As it is shown in figure 1, can by ICT-EIA method
Reduce the non-specific adsorption (background) of the antibody used, so multiple macromolecule physiologically active thing
The mensuration of the amol (zmol) below horizontal of matter may become (with reference to Hashida S, et al.,
Biotechnology Annual Review Vol.1, (1995) pp 403-451, Elsevier
Science Publishers B.V., Amsterdam).
" adiponectin " of the present invention refers to one of Adipocyte Factor from adipocyte secretion, is
The protein being made up of the amino acid of 244, be reported in blood with 3 aggressiveness (LMW:
Low-molecular-weight) as basic structure, exist 6 aggressiveness (MMW: middle-molecular-weihydroxyethyl), 18 gather
Several the polymer such as body (HMW: HMW).Its blood level with up to 5～
The concentration of 10 μ g/mL exists, and is had anti arteriosclerosis effect or insulin resistance to improve by report
The physiologically active of effect etc..Furthermore in recent years, these physiological action and the effect of multimeric structure
Being concerned, macromolecule polymer level or macromolecule polymer/total amount ratio are, in interior fat type
Obesity or diabetes, sugar tolerance, insulin resistance, coronary artery disease, metabolic syndrome
Furthermore Deng various morbid state by reduced training of have dinner therapy or operation etc., also show
Compare adiponectin total amount more relevant by force, show the importance that the molecule of adiponectin is the most quantitative.Furthermore,
By in the insulin resistance caused by obesity, also find out that the adiponectin of HMW is especially relevant.
It addition, according to the report inquired into for adiponectin and nephropathy, in adiponectin defect mouse,
The fusion that the podocytic process of the sertoli cell seeing kidney rises, confirms albuminuria.Also it is reported that, to
Supplement adiponectin at this, then see the normalization of the pathological tissue of kidney and the improvement of albuminuria,
Prompting adiponectin have effect for nephropathy protective possibility (Sharma Ketal:
Adiponectin regulates albumin uria and podocyte function in mice.J
Clin Invest 118 (5): 1645-1656,2008.).
Have again and know, ICT-EIA method measures main HMW adiponectin, measure a part
MMW adiponectin and unimolecule.But, although it is very weak reaction, but known also measures
LMW adiponectin.
" renal function " of the present invention refers to filtration and the re-absorption function of the glomerulus of kidney.
" evaluation index " of the present invention refers to that the adiponectin concentration in urine is dense divided by the adiponectin in blood
The numerical value of degree, such as, in the case of Healthy People, as seen in experimenter A and B, be 0.3 × 10-4
Below.On the other hand, it is renal dysfunction (microalbuminuria) in early days being prompted
In experimenter D, become 4.0 × 10-4.So, the evaluation index of the present invention becomes the number of more than 10 times
The difference of value, is the index of easily difference.This numerical value can be made to be multiplied by 10 again4, as integer, also
Evaluation transfiguration can be made easy.
[embodiment of the second of the present invention]
The embodiment of the second of the present invention relates to the prevention of renal function disease or methods for the treatment of
Invention, it is characterised in that be the degree of carrying out using above-mentioned evaluation index to carry out renal function disease
The method of evaluation method, also has, for there being the patient of renal dysfunction to carry out interventional therapy.
" degree of the carrying out evaluation method " of the present invention refers to the adiponectin concentration/blood in being urinated by mensuration
In the variation of ratio of adiponectin definitely evaluate experimenter renal function disease carry out situation
Method.Thus evaluation method, becomes definitely to carry out renal function disease for suppressing experimenter
Using or the interventional therapy of Life Guidance etc. of the therapeutic agent carried out and improve.
" interventional therapy " of the present invention refers to habits and customs based on flow process (-body movement of having dinner
Center) improve or known remedy for hyperlipemia, the taking of diabetes conditions therapeutic agent.
That is, instruct and will have dinner therapy or kinesiatrics is incorporated among habits and customs, carry out necessary heat and disappear
Take the control of heat, alleviate psychogenic stress, carry out deleting and alleviate sending out of diabetic keratopathy kidney diaseases
Relate in disease will be because of.Further, for the renal function disease patient of initial symptoms, as intervention
Therapy is containing the medicinal treatment of the numerical value of the evaluation index carried out for reducing the present invention.Use above-mentioned
Degree of the carrying out evaluation method of renal function disease, by starting interventional therapy in early days, suppression,
Avoid the migration to renal dysfunction of the severe degree of such as nephrosis etc., improve symptom,
Return healthy state.
Further, in the embodiment of the second of the present invention use term in, with first reality
The common term of mode of executing represents identical connotation.Therefore, mention the most especially.
[embodiment]
Next put off until some time later the bright present invention for embodiment, but the invention is not restricted to these.
[embodiment 1: by the mensuration of the adiponectin concentration in the blood of ICT-EIA method and in urine]
[(1) reagent]
[(a) antigen]
Standard recombinant-human adiponectin is from Bio Vendor Lab Med Inc company
(Palackecho, Czech Republic) buys.
[(b) antibody]
Monoclonal anti-human-adiponectin antibody (BAF1065 and MAB10651) is from R&D
Systems Inc. (MN, USA) buys.
[(c) buffer solution]
Buffer A: containing 0.1M NaCl, 0.01%BSA, 1mM MgCl2And 0.1%NaN3
0.01M sodium phosphate buffer (pH7.0),
Buffer B: 0.1M sodium phosphate buffer (pH6.0) containing 5mM EDTA,
Buffer solution C: containing 0.1M NaCl, 0.1%BSA, 1mM MgCl2And 0.1%NaN3
0.01M sodium phosphate buffer (pH7.0),
Buffer solution D: containing 0.4M NaCl, 0.1%BSA, 1mM MgCl2And 0.1%NaN3
0.05M sodium phosphate buffer (pH7.0),
Buffer solution E: the 0.01M sodium phosphate buffer (pH7.0) containing 0.1M NaCl
[(d) blood sample]
The blood serum sample of experimenter is taken a blood sample on an empty stomach 12 hours levees the most after the meal
(NIPRO, 22G seat is paid, Osaka), stand after 30 minutes in room temperature, with centrifuge (day
Vertical high speed centrifugal machine for minim CF16RXII, Hitachi's work machine, Tokyo) it is centrifuged 10 with 3000rpm
Minute, obtain serum.Serum is in negative 30 DEG C of freezen protective.
[(e) urine samples]
Urina sanguinis (10mL) to experimenter adds 5%BSA and 5%NaN of 1/50 capacity3,
Preserve to dialysis in 4 DEG C.Dialyse within 24 hours gather urine.
Dialysis uses dialysis cellulose tube (dialysis membrane UC8-32-25, the three pure medicines of light, Tokyo),
1.5ml is carried out for buffer solution (the 0.01M sodium phosphate buffer pH7.0 containing 0.1M NaCl)
The dialysis of above-mentioned urine samples.After dialysis, the weight of weighing dialysis urine, carry out by dilution ratio
Correction.Dialysis urinates immediately that dispensing is in micro-pipe (Safe-Lock Tubes 1.5ml), in-20 DEG C
Carry out freezen protective.Further, Concentrations is by (kreatinin-the test of commercial reagent box in Niao
WAKO and the pure medicine of light, Osaka) measure.
[(2) method]
Assay method uses method based on the ICT-EIA method described in non-patent literature 1.That is,
Prepare standard high score Ziren subsidiary for commercial ELISA Assay kit-adiponectin titer with slow
Rush 100 μ L of liquid D dilution or urine and serum buffer solution D are diluted 40,000 times
Enzyme is labeled antibody by 100 μ L, Xiang Qijia and capture is dissolved in buffer solution D with labeled antibody
Mixed liquor 100 μ L, in 4 DEG C of incubations 16 hours, make by enzyme be labeled antibody-adiponectin-
The capture immune complex of 3 compositions of labeled antibody is formed.It follows that react to this
Liquid adds the insoluble polystyrene bead of anti-DNP-IgG 1, makes reaction 0.5 hour, catches on pearl
Obtain immune complex.After this pearl is cleaned 2 times by buffer A (2mL), it is dissolved in buffering
The 2mMDNP-Lys (150 μ L) of liquid C, and make reaction 0.5 hour, make immunity multiple from pearl
Compound dissolution.After removing the insoluble polystyrene bead of anti-DNP-IgG, add strepto-to dissolution fluid
The insoluble polystyrene bead of Avidin 1, then make reaction 0.5 hour, make immune complex turn
Move on on the 2nd pearl.Reaction with pearl is all carried out under the vibration of 210 beats/min in 25 DEG C.Again
After pearl is cleaned 3 times with buffer solution C (2mL), use the β-D-pyrans half transferring on pearl
Lactoside (fluorogenic substrate；4MUG) (400 μ L), in 30 DEG C of incubations, adds 0.1M glycine
Sodium buffer solution (pH10.5) (2mL) and stop reaction after, use sepectrophotofluorometer
(F-2500, Hitachi) is measured.Further, use excitation wavelength 360nm wavelength of fluorescence
450nm, fluorescence intensity is with 10-8M4MU carries out (the Hashida Sand that converts as 100
Ishikawa E:Detection of one milliattomole of ferritin by novel and
Ultrasensitive enzyme immunoassay.J Biochem 108:960-964.1990.).
[(3) measurement result]
[(a) experimenter]
In order to evaluate as adiponectin in the urine of renal function index, screening has different sex, year
Age, the object of renal function, carry out in blood and the discussion of adiponectin in urine.Strong with having of youth
20～29 years old women A of the renal function of health, unsoundness renal function 40～49 years old women B,
With the age together as the eGFR (mL/min/1.73m of renal function presumption index2) see that reduction is inclined
To 50～59 years old male sex C, eGFR do not become C, but be prompted to be in microalbuminuria
Tendency, 60～69 years old obese males D of the reduction of the filter capacity of glomerulus are as object.
[(b) evaluation result]
First, Concentrations and age and property from the result, blood of general biochemical analysis
Do not calculate eGFR.Result is shown in table 1.Experimenter A is 97.4mL/min/1.73m2, tested
Person B is 107.2mL/min/1.73m2, experimenter C be 48.2mL/min/1.73m2, tested
Person D is 50.0mL/min/1.73m2。
[table 1]
| Experimenter | A | B | C | D |
| Sex | Female | Female | Man | Man |
| Age (year) | 22 | 40 | 58 | 64 |
| BMI(kg/m2) | 17.2 | 19.2 | 22.3 | 29.6 |
| Interior fat sectional area (cm2) | 34.8 | 56.6 | 50.4 | 118.7 |
| HbA1c (%) | 4.7 | 5.2 | 5.4 | 6.0 |
| Albumin (μ g/mg Cr) in urine | 3.88 | 1.29 | 4.79 | 33.6 |
| Serum creatinine (mg/dL) | 0.48 | 0.44 | 1.23 | 1.16 |
| eGFR(mL/min/1.73m2) | 97.4 | 107.2 | 48.2 | 50.0 |
| Serum adiponectin (μ g/mL) | 21.3 | 20.7 | 2.58 | 14.8 |
| Adiponectin (ng/mg Cr) in urine | 0.56 | 0.30 | 2.16 | 5.93 |
| In urine/blood in adiponectin ratio | 0.3×10-4 | 0.1×10-4 | 8.4×10-4 | 4.0×10-4 |
From the eGFR value of above-mentioned table 1, the renal function of experimenter C and D, as in figure 2 it is shown,
According to CKD Stage Classification, then corresponding to stage 3 (GFR30～50mL/min/1.73m2)
GFR moderate reduce.It addition, by albumin concentration in the urine of ICT-EIA method be, tested
Person A be 3.88 μ g/Cr, experimenter B be 1.29 μ g/Cr, experimenter C be 4.79 μ g/Cr,
Experimenter D is 33.6 μ g/Cr.Cannot be with single diagnosis, but the value of experimenter D is corresponding to micro-
The scope of amount albuminuria.
By the up to 21.3 μ g/mL in experimenter A of adiponectin concentration in the blood of ICT-EIA method,
Also 20.7 μ g/mL it are up in the same manner as A in experimenter B.But it is in experimenter C, low
In 2.58 μ g/mL and 4 μ g/mL, therefore show Hypoadiponectinemia.Experimenter D is
14.8 μ g/mL, have sex or age, underlying diseases, although without exercise habit, but also show height
Value.
In the urine measured in the same manner as blood, adiponectin concentration is in experimenter A
0.56ng/mgCr, is 0.30ng/mgCr in experimenter B.It addition, in the urine of experimenter C
Adiponectin concentration is 2.16ng/mgCr, compares experimenter A or B higher, and experimenter D is also same
Show the value of up to 5.93ng/mgCr sample.
It follows that obtain urinate relative to the adiponectin concentration in blood in the adiponectin concentration that leaches
Ratio time, experimenter A is 0.3 × 10-4, B be 0.1 × 10-4, in contrast to this, experimenter C
It is 8.4 × 10 with D-4With 4.0 × 10-4, show the value that decades of times is high.So, being subject to shown in Fig. 2
The stage of the renal function of examination person can with the evaluation index of the present invention (in urine/blood in adiponectin than) bright
Really distinguish, even corresponding to the object of the scope of microalbuminuria as experimenter D
Person, it is possible to by using this evaluation index to early diagnose, becomes to start interventional therapy in early days.
[embodiment 2: the existence pattern of the adiponectin in the blood of experimenter A～D and urine
Inquire into]
By the molecular sieve column of use Superde × 200, molecular weight differently classification separates dilution
The serum of experimenter A～D and urine, its classification separates and is measured by ICT-EIA method.Result
Knowing, it is several in the same manner as pattern in blood so far to there is pattern in the adiponectin of each object in serum
It is HMW adiponectin and MMW adiponectin.
[the existence pattern of the adiponectin in (a) urine]
Adiponectin in urine is as shown in table 2 below, at experimenter A, B of normal renal function
In, considerably less as absolute magnitude, but HMW, MMW, LMW detected in the same manner as so far
And unimolecule adiponectin.
[table 2]
As shown in Table 2 above, know in experimenter A, B especially, relative unimolecule adiponectin
Many.On the other hand know, as experimenter C, D equal LMW adiponectin of renal function reduction person
Peak account for most.It addition, can't detect HMW adiponectin in experimenter C.
[the existence pattern of the adiponectin in (b) blood]
There is pattern such as Fig. 3 (●) and advised in the adiponectin in the serum of experimenter A～D, several
It is HMW adiponectin and MMW adiponectin.The existence pattern of the adiponectin in urine shows significantly
Ground is different.Thus, measure this urine and neutralize the concentration of the HMW adiponectin in blood, take than (urine
Adiponectin concentration in middle adiponectin concentration/blood), then as more significant index, can become to evaluate kidney
Handicapped initial symptoms.
[embodiment 3: to the adiponectin value in the urine of index eGFR of renal function and albumin
The evaluation of the correlation of value]
[(a) correlation to eGFR]
In Gernral Check-up, measure the fat from the urine that experimenter 70 gathers based on embodiment 1
Connection element concentration (ng/mgCr) and albumin concentration (μ g/mgCre).It addition, simultaneously from being subject to
In the blood of examination person, Concentrations and age and sex calculate eGFR.
As a result, confirm between the albumin concentration in urine and eGFR to close less than the most relevant
System, but between adiponectin concentration and the eGFR in urine, as shown in Figure 4, confirm significantly
Negative dependency relation.Further, between adiponectin concentration (μ g/mL) and eGFR in blood,
Do not find significant dependency relation.
[(b) correlation to each stage (G1, G2, G3a) of eGFR]
From the eGFR value of the experimenter 70 of preceding paragraph, experimenter be categorized into shown in Fig. 2 is each
Stage.
To be categorized into stage G1 (more than 90), stage G2 (60～89), stage G3a (45～
59) in the urine of object adiponectin concentration (ng/mgCr) and urine in albumin concentration
(μ g/mgCre) is comprehensive.
Result is shown in Fig. 5 and 6.Relatively adiponectin concentration and urine in the urine between above-mentioned each stage
Middle albumin concentration, then in urine, adiponectin concentration is compared the G1 phase in the G2 phase and is shown the highest
Value.On the other hand, in urine, albumin concentration is interim at G3a, compares the G2 phase and shows significantly more
High value.
Above result shows that albumin concentration is more as nephropathy during adiponectin concentration compares urine in urine
Prediction index is useful.
[(c) be adiponectin concentration and albumin concentration in urine in the urine in each stage of eGFR
Relative value]
By dense for albumin in adiponectin concentration in the urine of the experimenter of the stage G1 of eGFR and urine
The median of degree is respectively as 1.Stage G2 and the experimenter of stage G3a is determined it follows that calculate
Urine in adiponectin concentration with urine in albumin concentration each become what times.
Result is shown in Fig. 7.That is, in stage G2, adiponectin relative value and white egg in urine in urine
White relative value ratio, shows the highest value.
Above result shows that in urine, adiponectin relative value is as the serviceability of nephropathy prediction index.
[industrial applicibility]
The evaluation index of the renal dysfunction of the present invention and its evaluation method with in the urine gathered
Present in the adiponectin concentration existed and the blood that simultaneously gathers adiponectin concentration be used for comment
The numerical value of valency index, grasps the initial stage function barrier of kidney in early days from the numerical value of the evaluation index obtained
The situation hindered becomes possibility.It addition, use the evaluation method of the present invention, build to Gernral Check-up
Application and the medical system of combination of interventional therapy become possibility.
Claims (3)
1. the evaluation index of renal function, it is characterised in that be with immune complex transfer assay side
The measured value of the adiponectin concentration in the urine that method (ICT-EIA method) measures joins divided by the fat in blood
The numerical value of element concentration.
2. the evaluation index of the renal function described in claim 1, wherein said adiponectin concentration is
HMW (HMW) adiponectin concentration.
3. the evaluation index of the renal function of claim 1 or 2, it is characterised in that be claim
The numerical value of 1 or 2 is multiplied by 104Numerical value.
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