CN113311172A - Application of urine parathyroid hormone as diabetic nephropathy DN (DN) prognostic diagnosis marker - Google Patents
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
- G01N2333/72—Assays involving receptors, cell surface antigens or cell surface determinants for hormones
- G01N2333/726—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/04—Endocrine or metabolic disorders
- G01N2800/042—Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/34—Genitourinary disorders
- G01N2800/347—Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Abstract
The invention discloses application of parathyroid hormone type 1 receptor PTH1R in urine to a kidney prognosis diagnostic marker of a patient with type 2 diabetes mellitus complicated with diabetic nephropathy. And the parathyroid hormone type 1 receptor PTH1R is applied to the preparation of the diabetic nephropathy DN prognostic marker. PTH1R levels were elevated in the urine of DN patients and were significantly correlated with 24 hour urine protein quantification, estimated glomerular filtration rate eGFR, slope of eGFR decline, and the extent of pathological injury including tubular atrophy, interstitial fibrosis, and glomerulosclerosis. DN patient urine PTH1R level is an independent risk factor for the patient to end-of-kidney progression (ESRD), and may improve the prognosis predictability of DN patients.
Description
Technical Field
The invention relates to the field of gene biological preparations, in particular to a new application of a urine parathyroid hormone type 1 receptor PTH1R as a kidney long-term prognosis diagnostic marker of a diabetic nephropathy DN patient.
Background
With the improvement of the life style of people in China, the change of the dietary structure and the aging of population, the incidence of diabetes in China is continuously increased, and Diabetic Nephropathy (DN) is one of the most common microvascular complications of diabetes. Patients with DN enter End Stage Renal Disease (ESRD) more readily than non-diabetic renal disease (NDKD) patients. Yet there is still a lack of noninvasive biomarkers that can predict early renal function progression in DN patients.
Chronic Kidney Disease (CKD) patients can develop a series of mineral metabolism disorders after entering CKD3 stage, which can be manifested by decreased blood calcium levels, increased blood phosphorus levels, increased parathyroid hormone (PTH) expression and decreased 25 hydroxyvitamin D expression. However, studies have shown that the diabetic patients can develop mineral metabolism disorder when complications do not occur, and the phenomenon can be aggravated when kidney diseases are combined. Furthermore, DN patients may develop more severe mineral metabolism disorders earlier than NDKD patients. Thus, in DN patients, mineral metabolites are likely to be early biomarkers for predicting renal function progression.
Parathyroid hormone (PTH), a typical endocrine hormone, is a key factor in regulating blood calcium levels and is essential for maintaining body ion homeostasis and skeletal health. The detection kit of the whole parathyroid hormone (iPTH) commonly used at present has cross reaction with a PTH degradation fragment PTH (7-84), so that the measured iPTH overestimates the level of the whole parathyroid hormone PTH (1-84), namely iPTH, and the iPTH cannot well predict the kidney fate. Parathyroid hormone type 1 receptor (PTH1R) is the receptor for PTH (1-84) and PTH (7-84), expressed primarily in the bone and kidney. PTH specifically binds to PTH1R, a B-class G protein-coupled receptor (GPCR) family member highly expressed in bone and kidney cells, and activates downstream signaling pathways to regulate calcium and phosphorus metabolism in vivo. Therefore, PTH1R is a recognized therapeutic target for osteoporosis, and related drugs have been clinically used. PTH1R mediates the hypercalcemic effects of PTH (1-84), while PTH (7-84) induces PTH1R degradation through ubiquitination. Thus, it is theorized that the level of PTH1R in the tissue better reflects the true concentration of PTH (1-84) in the blood.
In addition, studies have shown that PTH1R phosphorylation-deficient mice can exhibit PTH-induced decrease in glomerular filtration rate, and systemic injection of PTH1R cDNA plasmid in animal models can reduce renal vascular resistance and plasma renin concentrations, suggesting that PTH1R may be involved in the regulation of renal hemodynamics. In summary, it is important to identify the relationship between PTH1R and renal function progression and renal outcome in DN patients.
Disclosure of Invention
The invention aims to provide application of urine parathyroid hormone type 1 receptor in preparation of diabetic nephropathy DN prognostic markers, so as to solve the problem that DN still lacks appropriate renal prognostic judgment markers in the prior art.
In order to achieve the purpose, the invention adopts the technical scheme that:
use of the parathyroid hormone type 1 receptor PTH1R as a DN prognostic marker for diabetic nephropathy.
The parathyroid hormone type 1 receptor PTH1R is applied to a detection preparation of a diabetic nephropathy DN prognostic marker.
The parathyroid hormone type 1 receptor PTH1R is capable of increased expression in the urine of patients with diabetic nephropathy DN. The larger the relative amount of expression of parathyroid hormone type 1 receptor PTH1R, the higher the urine protein quantification at 24 hours, the lower the eGFR and the higher the eGFR decrease slope, indicating that the prognosis of diabetic nephropathy DN is poor.
The parathyroid hormone type 1 receptor PTH1R is significantly related to 24-hour urinary protein quantification, eGFR decline rate and pathological damage degree including tubular atrophy, interstitial fibrosis, glomerulosclerosis and the like of diabetic nephropathy DN patients.
The parathyroid hormone type 1 receptor PTH1R is used as a prognostic marker, and urine ELISA detection is carried out to obtain the relative quantity of urine PTH1R expression.
The parathyroid hormone 1 type receptor PTH1R is used as a prognostic marker, and urine ELISA detection is carried out to obtain the relative quantity of urine PTH1R expression; PTH1R can be popularized to clinical application as DN prognostic marker index of diabetic nephropathy, and although a commercial ELISA kit exists at present, the kit is not clinical diagnosis grade. According to the invention, the future clinical detection is not detection by ELISA, but can be a radioimmunoassay, so that the scheme for preparing the reagent can be various.
Compared with the prior art, the invention has the following beneficial effects:
the invention is proved by research: PTH1R was expressed in the urine of DN patients and was significantly correlated with 24 hour urinary protein quantification, eGFR decline rate, and the extent of pathological injury including tubular atrophy, interstitial fibrosis, and glomerulosclerosis in DN patients.
The inventor finds that PTH1R is increased in urine of DN patients by detecting urine of DN patients, and can effectively improve the prediction capability of DN patient prognosis.
Drawings
FIGS. 1A-1C Pearson correlation analysis show the correlation of ELISA-detected urine PTH1R levels with DN patient renal function index (FIG. 1A)24 hour urine protein quantification, (FIG. 1B) glomerular filtration rate eGFR, (FIG. 1C) glomerular filtration rate decline slope eGFR-slope.
Fig. 2A-2CPearson correlation analysis shows the correlation between urine PTH1R levels and DN patient renal pathology score (fig. 2A) renal interstitial fibrosis, (fig. 2B) tubular atrophy, (fig. 2C) glomerulosclerosis.
FIG. 3 the Kaplan-Meier curve shows the results of the kidney survival curves for different urine PTH1R levels.
Figure 4 urine PTH1R levels improved the predictive power of DN patient prognosis. In combination with urine PTH1R levels, the combination of 24 hour urine protein quantification and eGFR is significantly better than the predictive ability of the combination of 24 hour urine protein quantification and eGFR.
Detailed Description
The present invention will be further described with reference to the following examples. When the significance of the data normality test result is greater than 0.05, the data obeys normal distribution, and Pearson correlation analysis is continuously carried out;
example 1
Application of urine PTH1R as DN prognosis marker
Detection of urinary PTH1R levels:
1) detection of urine PTH1R by ELISA method: the human PTH1R kit is purchased from LifesPan company, 190 cases of DN patients (total) urine and the kit are recovered to room temperature according to the kit specification, a standard substance and a sample are diluted, the sample is added and incubated for 2h at room temperature, the washing solution is washed for 3 times, an HRP cross-linking antibody is added, the incubation for 2h at room temperature is carried out, the washing solution is washed for 3 times, a developing solution is added, the incubation for 20min in dark at room temperature is carried out, a stop solution is added, an ELISA reader is used at 450nm, the O.D. value is corrected and read at 540nm or 570nm, a standard curve is obtained by using a log/log algorithm, and the PTH1R concentration of each sample is obtained by calculation.
2) And (3) detecting urinary creatinine: the creatinine kit is purchased from bioassay company, samples and standard substances are added according to the kit specification, developing solution is added, O.D. values of 0min and 5min are read at 510nm by an enzyme-linked immunosorbent assay, and the creatinine value of the sample is obtained by using a linear formula.
3) Urine levels of PTH1R were corrected for urine creatinine levels, resulting in relative amounts of urine PTH1R expression, and log transformation [ uPTH1R/Cr (log)10)]。
This example found that PTH1R was elevated in the urine of DN patients and was significantly associated with 24 hour urine protein quantification, eGFR decline rate, and the extent of pathological injury including renal interstitial fibrosis, renal tubular atrophy, and glomerular sclerosis; urinary PTH1R levels were effective in improving the predictive power of DN patient prognosis. As shown in fig. 4.
In combination with urine PTH1R levels, the combination of 24 hour urine protein quantification and eGFR is significantly better than the predictive ability of the combination of 24 hour urine protein quantification and eGFR.
In review, this example shows that urine PTH1R is of great significance in the prognostic evaluation of DN patients.
The above description is only of the preferred embodiments of the present invention, and it should be noted that: it will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the principles of the invention and these are intended to be within the scope of the invention.
Claims (5)
1. Use of parathyroid hormone type 1 receptor PTH1R as a renal prognostic diagnostic marker in patients with type 2 diabetes mellitus complicated with diabetic nephropathy.
2. Use according to claim 1, characterized in that: the parathyroid hormone type 1 receptor PTH1R is applied to the preparation of a diabetic nephropathy DN prognostic marker.
3. Use according to claim 1 or 2, characterized in that: the parathyroid hormone type 1 receptor PTH1R is capable of increased expression in the urine of patients with diabetic nephropathy DN.
4. Use according to claim 1 or 2, characterized in that: the parathyroid hormone type 1 receptor PTH1R was significantly associated with urinary protein quantification, estimated glomerular filtration rate eGFR, eGFR decline slope, and pathological damage extent including tubular atrophy, interstitial fibrosis, and glomerulosclerosis in diabetic nephropathy DN patients.
5. Use according to claim 1 or 2, characterized in that: the parathyroid hormone type 1 receptor PTH1R is used as a prognostic marker, and urine ELISA detection is carried out to obtain the relative quantity of urine PTH1R expression.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103293250A (en) * | 2013-05-13 | 2013-09-11 | 罗国安 | Diabetic nephropathy diagnostic kit and application thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103293250A (en) * | 2013-05-13 | 2013-09-11 | 罗国安 | Diabetic nephropathy diagnostic kit and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| ARANTXA ORTEGA等: "Parathyroid Hormone-Related Protein Is a Hypertrophy Factor for Human Mesangial Cells: Implications for Diabetic Nephropathy", 《JOURNAL OF CELLULAR PHYSIOLOGY》 * |
| MONTSERRAT ROMERO等: "Novel Role of Parathyroid Hormone-Related Protein in the Pathophysiology of the Diabetic Kidney: Evidence from Experimental and Human Diabetic Nephropathy", 《JOURNAL OF DIABETES RESEARCH》 * |
| RICARDO J. BOSCH等: "A Transgenic Mouse Model for Studying the Role of the Parathyroid Hormone-Related Protein System in Renal Injury", 《JOURNAL OF BIOMEDICINE AND BIOTECHNOLOGY》 * |
| 朱珍等: "甲状旁腺激素相关蛋白在糖尿病肾病中的研究进展", 《中华内分泌代谢杂志》 * |
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