Application of MMP19 as biomarker related to early kidney injury in early diagnosis of kidney injury
A biomarker, MMP19 technology, applied in the application field of early diagnosis of kidney injury, can solve the problem of not being able to detect early kidney injury and other problems
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Embodiment 1
[0082] Acute Kidney Injury (AKI) Mice Experiment
[0083] A mouse model of renal ischemia-reperfusion injury (IRI)
[0084] The same batch of healthy male wild-type C57BL / 6 mice about 8 weeks was selected and randomly divided into 5 groups with 4 mice in each group, control group, ischemia-reperfusion. Autoclave all surgical instruments and dehair the mouse abdomen in advance.
[0085] Preparation of anesthetics: Mix 25 g of tribromoethanol with 15.5 mL of tert-amyl alcohol to prepare a 1600 mg / mL concentrated stock solution of tribromoethanol, store it in the dark after complete dissolution, and prepare a 20 mg / mL working solution (diluted with normal saline) before use.
[0086] Anesthesia: The mice were anesthetized by intraperitoneal injection of 20 mg / mL tribromoethanol working solution at a dose of 350 mg / kg (17.5 μl / g mouse body weight), and the toes of the mice were lightly clamped with tweezers to determine whether they entered a state of deep anesthesia.
[0087] O...
Embodiment 2
[0093] Acute Kidney Injury Clinical Sample Detection Experiment
[0094] Serum and urine samples were collected from patients with acute kidney injury. Morning urine / random urine was collected, and the urine samples were centrifuged at low temperature (4°C, 3000rpm, 10min). The urine supernatant was collected, 1mL / tube, and frozen at -80°C. In addition, serum samples from patients with renal biopsy were collected on the same day, and one tube of biochemical serum coagulant vacuum blood collection tube was used for sample collection. Immediately after blood collection, invert and mix 8 times, and stand for 15-20 minutes for low temperature centrifugation (4°C, 3000 rpm, 10 min). The supernatant was collected and aliquoted into 20 μL / tube and frozen at -80°C. A sample test is performed once a month.
[0095] The samples clinically diagnosed as acute kidney injury were collected, and the inclusion criteria were as follows: 1. Within 48 hours, the average index of serum crea...
Embodiment 3
[0106] Chronic kidney injury (CKD) mouse experiment
[0107] Unilateral ureteral obstruction (UUO) model
[0108] The same batch of healthy male wild-type C57BL / 6 mice for about 8 weeks was selected and randomly divided into 4 groups, the control group, the third day of modeling (UUO-3d) and the seventh day of modeling (UUO-7d). On the 14th day of modeling (UUO-14d), there were 4 rats in the control group and 6 rats in the model group. Autoclave all surgical instruments and dehair the mouse abdomen in advance.
[0109] Preparation of anesthetics: Mix 25 g of tribromoethanol with 15.5 mL of tert-amyl alcohol to prepare a 1600 mg / mL concentrated stock solution of tribromoethanol, store it in the dark after complete dissolution, and prepare a 20 mg / mL working solution (diluted with normal saline) before use.
[0110] Anesthesia: The mice were anesthetized by intraperitoneal injection of 20 mg / mL tribromoethanol working solution at a dose of 350 mg / kg (17.5 μl / g mouse body wei...
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