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Application of cryab in the detection of acute kidney injury and its detection kit

A detection kit and kidney injury technology, applied in the field of medicine, can solve the problems of kidney injury grading, early detection of AKI, etc., and achieve the effect of reducing chronic kidney disease complications and avoiding or chronic kidney disease complications

Active Publication Date: 2022-02-11
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is: how to solve the problems in the prior art that AKI cannot be detected early and the degree of renal damage suffered by the kidney cannot be graded

Method used

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  • Application of cryab in the detection of acute kidney injury and its detection kit
  • Application of cryab in the detection of acute kidney injury and its detection kit
  • Application of cryab in the detection of acute kidney injury and its detection kit

Examples

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Effect test

Embodiment 1

[0022] Example 1. To demonstrate αB-crystallin as a sensitive and early biomarker of AKI, we used a mouse model of renal ischemia / reperfusion (I / R). 8-12 week old male C57BL / 6 mice were used throughout the study. The mice were anesthetized by intraperitoneal injection of 50-60 mg / kg pentobarbital and subcutaneous injection of 50 ug / kg buprenorphine, and placed on a constant temperature monitoring surgical blanket to balance their body temperature to 36.5-37°C. A 1-1.5 cm longitudinal surgical incision was made on the left side of the mouse from the abdomen to the 1 / 3 of the spine (near the spine); a 1-1.5 cm transverse incision was made on the right side along the ribs at the same position. Enter the abdominal cavity layer by layer, squeeze out the kidneys with cotton swabs, and bluntly separate the tissues around the renal pedicles. After exposing the renal blood vessels, put the kidneys back into the abdominal cavity. After 25 minutes, the color of the kidney changed from b...

Embodiment 2

[0024] Example 2. In all groups, renal function was assessed by measuring serum urea nitrogen, creatinine. For the detection of serum urea nitrogen, mix the mouse serum sample (standard control and blank deionized water control at the same time) with BUN color Reagent and BUN Acid Reagent, heat at 95°C for 10 minutes, place in ice water for 3-5 minutes, and mix thoroughly. After uniformity, detect the absorbance value at 520nm, draw a standard curve, and calculate the serum urea nitrogen value of each sample.

[0025] For serum creatinine detection, configure standard products 0, 66.3, 132.6, 198.9, 265.2 μM. Prepare 200ul working solution 0.96% sodium hydroxide: 0.83% picric acid=1:1, bathe in 37℃ water for 3 minutes, add 20μL standard or sample, time 20 seconds to read the absorbance value at 510nm, and then time 60 seconds to read the second time Absorbance value, draw a standard curve, and calculate the serum creatinine of the sample.

[0026] To investigate whether CryA...

Embodiment 3

[0027] Example 3. Cell protein extraction: collect cells (35mm culture dish as an example), collect the culture medium and centrifuge at 1000g for 5 minutes, discard the supernatant, wash the cells with 1×PBS and collect them in the same centrifuge tube and centrifuge at 1000g for 5 minutes, discard the supernatant . Add 100 μl protein lysate to fully cover all adherent cells, scrape the cells and lysate to one side with a clean scraper and transfer to the above centrifuge tube, lyse for a while, centrifuge at 15000g for 5 minutes to collect the supernatant for later use.

[0028]Determination of protein concentration and western blot analysis: the sample was diluted 10 times, and the standard was prepared according to the above method. Mix and prepare the working solution according to the ratio of A solution and B solution 50:1, add 200 μL of the working solution to each well of the 96-well plate, and add 10 μL each of the standard product and the diluted sample, repeat 2 pai...

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Abstract

The invention discloses the application of CRYAB in the detection of acute kidney injury and a detection kit. αB-crystallin can be used as a detection marker for the detection of acute kidney injury. Aiming at the discovery, a corresponding detection kit is made. The present invention helps to solve the existing problem in the field of medicine, that is, the inability to detect the severity of acute kidney injury at an early stage, so as to treat the patient with effective therapy in time.

Description

technical field [0001] The invention belongs to the field of medicine, and specifically relates to the use of αB crystallin in detecting ischemia-reperfusion kidney injury and its degree. Background technique [0002] Acute kidney injury (AKI) is a common clinical kidney disease with high morbidity and mortality caused by different reasons. In China, there are 4-6 million AKI incidences and 300,000-500,000 related deaths every year. At present, there are only adjuvant treatment methods such as dialysis for AKI, and there are no effective early diagnosis methods and therapeutic drugs. Acute kidney injury first appears in proximal tubular epithelial cells and manifests as various forms of cell death. So far, there is no effective drug for the treatment of acute kidney injury, and the clinical treatment mostly adopts supportive care. Therefore, early diagnosis and early treatment are the best strategies to prevent and treat acute kidney injury. [0003] Currently available t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/58
CPCG01N33/6893G01N33/581G01N33/543G01N2800/347G01N2800/56
Inventor 娄强侯贝贝
Owner HENAN UNIVERSITY
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