Application of Dectin-1 as biomarker in preparation of renal fibrosis diagnostic reagent

Abstract

The invention discloses application of Dectin-1 as a biomarker in preparation of a renal fibrosis diagnostic reagent or kit, and belongs to the field of biological medicine. And Dectin-1 is a member of a pattern recognition receptor C-type lectin family. According to the invention, Dectin-1 immunofluorescent staining is carried out on kidney tissues of patients with chronic kidney diseases, and observation shows that the number of Dectin-1 + cells in fibrosis kidneys is more. Dectin-1 immunofluorescence staining is further carried out in the kidney tissue of the patient with the IgA nephropathy, and it is found that the number of Dectin-1 + cells in the kidney tissue of the patient with the IgA nephropathy is in positive correlation with the renal fibrosis degree. The results show that the reagent for detecting the expression level of the Dectin-1 in the kidney cells can be used for preparing the kit for diagnosing the kidney fibrosis, and the diagnosis result has high sensitivity and specificity.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of Dectin-1 as a biomarker in the preparation of a diagnostic reagent for renal fibrosis. Background Art

[0002] Chronic kidney disease (CKD) has become a global public health problem, with a global prevalence of approximately 10%. It is projected to become the fifth leading cause of death worldwide by 2040. Regardless of the cause, nearly all types of CKD eventually develop into renal fibrosis, characterized by inflammatory cell infiltration, fibroblast activation and proliferation, excessive extracellular matrix deposition, tubular atrophy, and microvascular rarefaction.

[0003] The immune response plays a crucial role in renal fibrosis. Pattern recognition receptors expressed by innate immune cells are activated upon renal injury, triggering signal transduction and regulating the production of inflammatory mediators, thereby promoting renal fibrosis. Therefore, detection of these pattern recognition receptors and signaling molecules can help clarify the progression of renal fibrosis.

[0004] Dectin-1, a member of the C-type lectin family of pattern recognition receptors, is encoded by the CLEC7A gene and is primarily expressed on myeloid-monocytic cells such as macrophages. Initial studies suggested that Dectin-1 could recognize β-glucans in fungal cell walls, thereby initiating an antifungal immune response. However, recent studies have shown that Dectin-1 is also involved in non-pathogen-mediated sterile inflammation, such as autoimmune diseases, allergies, and cancer. However, whether Dectin-1 is involved in the progression of renal fibrosis in CKD patients remains unknown. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for preparing a diagnostic reagent for renal fibrosis as a biomarker, so as to solve the problems existing in the prior art. The present invention found that after immunofluorescence staining of Dectin-1, Dectin-1 + The number of cells was positively correlated with the degree of renal fibrosis.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The present invention provides the use of Dectin-1 as a biomarker in the preparation of a diagnostic reagent or kit for renal fibrosis.

[0008] Preferably, the renal fibrosis is renal fibrosis caused by chronic kidney disease or IgA nephropathy.

[0009] The present invention provides use of a reagent for detecting Dectin-1 levels in the preparation of a renal fibrosis diagnosis product.

[0010] Optionally, the reagent is an antibody that specifically binds to Dectin-1.

[0011] Preferably, the renal fibrosis is renal fibrosis caused by chronic kidney disease or IgA nephropathy.

[0012] Preferably, the Dectin-1 level is the expression level of Dectin-1 in renal tissue cells.

[0013] Preferably, in the renal tissue of the patient to be tested, there are a large number of cells with high Dectin-1 expression levels, indicating that the patient to be tested has a high risk of suffering from renal fibrosis.

[0014] Preferably, the renal fibrosis diagnostic product comprises a kit.

[0015] The present invention also provides a renal fibrosis diagnosis kit, comprising an antibody that specifically binds to Dectin-1.

[0016] Preferably, the renal fibrosis is renal fibrosis caused by chronic kidney disease or IgA nephropathy.

[0017] The present invention discloses the following technical effects:

[0018] The present invention provides the use of Dectin-1 of the pattern recognition receptor C-type lectin family as a biomarker in the preparation of a reagent for diagnosing renal fibrosis. The present invention performs Dectin-1 immunofluorescence staining on renal tissue of patients with chronic kidney disease and observes that Dectin-1 is expressed in fibrotic kidneys. + Further Dectin-1 immunofluorescence staining was performed in the renal tissues of patients with IgA nephropathy, and it was found that Dectin-1 + The number of cells is positively correlated with the degree of renal fibrosis. These results indicate that a reagent based on the detection of Dectin-1 expression levels in renal cells can be used to prepare a kit for diagnosing renal fibrosis, with high sensitivity and specificity. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0020] Figure 1 Representative immunofluorescence staining images of renal tissue from patients with various types of chronic kidney disease; red fluorescence represents Dectin-1, blue fluorescence represents DAPI, scale bar: 50 μm;

[0021] Figure 2 Representative images of renal tissues from patients with IgA nephropathy stained with HE, PAS, PASM, and Masson staining; scale bar: 50 μm;

[0022] Figure 3 Representative images of immunofluorescence staining of renal tissue from patients with IgA nephropathy, where red fluorescence represents Dectin-1 and blue fluorescence represents DAPI. Scale bar: 50 μm.

[0023] Figure 4 Dectin-1 for each high-power field of view + Cell count statistics; **** indicates P < 0.0001 compared with the T0 group;

[0024] Figure 5 Dectin-1 for each high-power field of view + Correlation analysis chart between cell count and blood creatinine, blood urea nitrogen and urine protein;

[0025] Figure 6 Dectin-1 for each high-power field of view + Correlation analysis between cell count and degree of renal fibrosis, serum albumin and glomerular filtration rate;

[0026] Figure 7 Dectin-1 for each high-power field of view + Receiver operating characteristic (ROC) curve of cell number for differentiating renal fibrosis T0 and T1 in patients with IgA nephropathy. DETAILED DESCRIPTION

[0027] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0028] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. The intermediate value within any stated value or stated range, and each smaller range between any other stated value or intermediate value within the stated range, is also encompassed within the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0029] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0030] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be illustrative only.

[0031] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0032] Example 1

[0033] The present invention performed Dectin-1 immunofluorescence staining on renal tissues of patients diagnosed with diabetic nephropathy, hypertensive nephropathy, lupus nephritis, minimal change disease, membranous nephropathy and mesangial proliferative glomerulonephritis by renal puncture biopsy at the First Hospital of Zhejiang University (Dectin-1 antibody was purchased from Abcam, catalog number ab140039). The degree of renal tissue fibrosis was assessed by Masson staining, and the renal interstitial fibrosis of the above CKD patients was not observed as a control. We found that Dectin-1 + Cells accumulate more in the fibrotic kidneys of CKD patients ( Figure 1 ).

[0034] Next, we enrolled 84 patients diagnosed with IgA nephropathy by renal puncture biopsy at the First Hospital of Zhejiang University. Exclusion criteria included: patients with other renal diseases; those who had received immunosuppressive therapy before diagnosis; renal transplant recipients; and those with active infection. According to the Oxford classification, patients with tubular atrophy or interstitial fibrosis ≤ 25% were defined as the T0 group, and patients with tubular atrophy or interstitial fibrosis 26%-50% were defined as the T1 group. We stained renal tissue sections of patients with IgA nephropathy and found that the tubular atrophy and interstitial collagen deposition in the T1 group were more obvious than those in the T0 group ( Figure 2 ).

[0035] Dectin-1 immunofluorescence staining showed that the renal tissue of patients in the T1 group had a significant increase in Dectin-1 in each high-power microscope field. + The number of cells was significantly increased compared with the T0 group ( Figure 3 and Figure 4 ).

[0036] Through correlation analysis, we found that Dectin-1 + The cell count was positively correlated with serum creatinine, blood urea nitrogen, urine protein and renal fibrosis in patients with IgA nephropathy, but negatively correlated with serum albumin and glomerular filtration rate ( Figure 5 and Figure 6 ).

[0037] The ROC curve of Dectin-1 as a biomarker for detecting the degree of renal fibrosis T0 and T1 in patients with IgA nephropathy is shown as follows: Figure 7 As shown, the area under the curve (AUC) is 0.94, indicating that the Dectin-1 provided by the present invention has excellent specificity and sensitivity for diagnosing renal fibrosis.

[0038] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. Application of Dectin-1 as a biomarker in the preparation of diagnostic reagents or kits for renal fibrosis.

2. The use according to claim 1, characterized in that The renal fibrosis is renal fibrosis caused by chronic kidney disease or IgA nephropathy.

3. Application of reagents for detecting Dectin-1 levels in the preparation of renal fibrosis diagnostic products.

4. The use according to claim 3, characterized in that The reagent is an antibody that specifically binds to Dectin-1.

5. The use according to claim 3, characterized in that The renal fibrosis is renal fibrosis caused by chronic kidney disease or IgA nephropathy.

6. The use according to claim 3, characterized in that The Dectin-1 level refers to the expression level of Dectin-1 in renal tissue cells.

7. The use according to claim 6, characterized in that In the renal tissue of the patient to be tested, there are a large number of cells with high Dectin-1 expression levels, indicating that the patient to be tested has a high risk of suffering from renal fibrosis.

8. The use according to claim 3, characterized in that The renal fibrosis diagnostic product includes a kit.

9. A renal fibrosis diagnostic kit, characterized in that: Included are antibodies that specifically bind to Dectin-1.

10. The kit according to claim 9, wherein The renal fibrosis is renal fibrosis caused by chronic kidney disease or IgA nephropathy.

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