Application of secretory leukocyte protease inhibitor (SLPI) as prognostic marker of diabetic nephropathy (DN)

A technology for protease inhibition and diabetic nephropathy, which is applied in disease diagnosis, analytical materials, biological testing, etc., can solve the problem of DN lacking renal prognosis markers and achieve the effect of predicting prognosis

Active Publication Date: 2020-08-11
蒋松
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  • Abstract
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  • Application Information

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Problems solved by technology

[0004] The present invention provides the use of secretory leukocyte protease inhibitor SLPI in the preparation of diabetic neph

Method used

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  • Application of secretory leukocyte protease inhibitor (SLPI) as prognostic marker of diabetic nephropathy (DN)
  • Application of secretory leukocyte protease inhibitor (SLPI) as prognostic marker of diabetic nephropathy (DN)
  • Application of secretory leukocyte protease inhibitor (SLPI) as prognostic marker of diabetic nephropathy (DN)

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Embodiment 1

[0022] A methodology for screening prognostic markers by combining transcriptomic profiling and clinical indicators

[0023] (1) Obtaining the Discovery Queue

[0024] The cohort used for transcriptome sequencing in the present invention comes from the prospective DN cohort of Nanjing General Hospital and the National Clinical Research Center for Kidney Diseases. Among them, DN patients were enrolled in the following conditions: a) age ≥ 18 years old; b) type 2 diabetes mellitus (WHO diagnostic criteria) with DN patients who were admitted to the National Clinical Research Center for Kidney Diseases in Nanjing General Hospital of Nanjing Military Command and diagnosed by renal biopsy , to exclude other concomitant renal diseases; c) sign the informed consent form; d) have microdissection of renal tissue on the basis of complete routine pathological examination; e) follow-up for at least 1 year, at least 2 eGFR follow-up points per year; co-enrolled DN There were 61 patients. ...

Embodiment 2

[0030] Application of SLPI as a prognostic marker for DN

[0031] (1) RT-PCR

[0032] Paraffin wax specimens of kidney tissue from patients with DN were sliced ​​into LEICA laser micro-separation special sections, freshly prepared for dewaxing in xylene, dehydrated with gradient alcohol, and separated by laser micro-separation of renal tubular tissue, and the tubular tissue was extracted with FFPE RNA extraction kit purchased from QIAGEN The total RNA was determined using a two-step method to determine the purity and concentration of RNA: Step 1 reverse transcription, reverse transcription of mRNA to cDNA according to the kit instructions, the reaction conditions are: 25°C, 10min, 37°C, 120min, 85°C, 5min; step 2: real-time PCR reaction, using SLPI and 18s as primers, and carried out on a real-time fluorescent quantitative PCR instrument. Reaction conditions: 95°C, 30s, 95°C, 5s, 60°C, 34s, 40 cycles, 95 ℃, 15s, 60℃, 1min, 95℃, 15s, record and observe the amplification curve ...

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Abstract

The invention discloses an application of a secretory leukocyte protease inhibitor (SLPI) as a prognostic marker of diabetic nephropathy (DN). Researches prove that the expression of the SLPI in renaltubular epithelial cells of a DN patient is increased, the expression of the SLPI in urine is also increased, the expression of the SLPI is significantly related to the decrease slope of the glomerular filtration rate (eGFR) of the DN patient, and the prognosis of the patient can be predicted. By detecting kidney tissue sections and urine of the DN patient, the inventor finds that the mRNA and protein levels of SLPI in renal tubules of the DN patient are remarkably increased, the expression level of SLPI in urine is increased in parallel with the protein level of the kidney, and prognosis ofthe DN patient can be effectively predicted and is superior to that of serum creatinine or proteinuria.

Description

technical field [0001] The invention belongs to the technical field of genetic biological preparations, and particularly relates to the application of secreted leukocyte protease inhibitor SLPI in the field of diabetic nephropathy DN. Background technique [0002] Diabetic nephropathy (DN) is one of the most common microvascular complications of diabetes. With the increasing prevalence of diabetes in my country, the incidence of DN also increases. Research shows that DN has surpassed glomerulonephritis and has become the primary cause of chronic kidney disease (CKD) in hospitalized patients in my country. Survey data from the United States in the past 20 years shows that under the background of effective control of important complications of diabetes, such as cardiovascular and cerebrovascular events, the incidence of end-stage renal disease (ESRD) caused by diabetes is not obvious. The diagnosis of DN still relies on clinical diagnosis and pathological diagnosis, and the ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2333/8107G01N2800/34G01N2800/52
Inventor 蒋松李申秦卫松刘志红
Owner 蒋松
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