Application of small molecule polypeptide kp-1 in preparing medicine for treating chronic kidney disease
A technology for small molecule peptides and chronic kidney disease, which is used in drug combinations, medical preparations containing active ingredients, and pharmaceutical formulations.
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Embodiment 1
[0024] Example 1: Screening of KP-1 Small Molecule Peptides
[0025] 1. Experimental materials:
[0026] Cells: Rat kidney fibroblasts (NRK-49F).
[0027] Culture medium: DMEM / F12 (1:1) culture medium containing 10% FBS.
[0028] Culture conditions: 37 o C contains 5% CO 2 incubator.
[0029] Small molecule peptide library: 18 peptide sequences were designed by computer, each with an average length of 30 amino acids, which were artificially synthesized by Nanjing GenScript Biotechnology Co., Ltd. to build a small molecule peptide library.
[0030] 2. Experimental treatment:
[0031] (I) The cultured rat kidney fibroblasts were mixed with 1.5×10 6 They were planted in 6-well cell culture plates, cultured for 1 day, starved for 24 hours, and given 18 different small molecule peptides (10 µg / ml) 1 hour in advance. Then stimulated with TGF-β recombinant protein (2ng / ml), collected the protein 48 hours later, and carried out Western blot experiments to screen the peptides th...
Embodiment 2
[0039] Example 2: Distribution of FITC-labeled KP-1 in various organs in UUO mice
[0040] 1. Experimental animals: C57 mice, male, weighing 20-22g, SPF grade.
[0041] The animals were first weighed and numbered, and 2 healthy mice weighing 20-22 g were selected and divided into UUO and UUO+KP-1.
[0042] 2. Experimental grouping
[0043] 1) UUO mice: at room temperature, anesthetize the mouse with 1% pentobarbital sodium at 5ml / kg body weight, select the left abdomen 2-3cm as the incision; after local disinfection, cut the skin, subcutaneous, muscle layer and For the peritoneum, after finding the left ureter, ligate the upper 1 / 3 of the ureter and suture layer by layer. After local disinfection, check the marks and place them in corresponding mouse cages.
[0044] 2) UUO+KP-1 mice: anesthetize and disinfect as above. The skin, subcutaneous layer, muscle layer and peritoneum were cut layer by layer. After finding the left ureter, the upper 1 / 3 segment of the ureter was li...
Embodiment 3
[0049] Example 3: Inhibitory effect of small molecule polypeptide KP-1 on renal fibrosis in mouse UUO model
[0050] 1. Experimental animals: BALB / C mice, male, weighing 20-22g, SPF grade.
[0051] The animals were first weighed and numbered, and 12 healthy mice weighing 20-22 g were selected and randomly divided into 3 groups, 6 in each group. Including sham operation group, model control group and medication group.
[0052] 2. Experimental grouping
[0053] 1) Sham operation group: After mice were anesthetized with 1% sodium pentobarbital at 5 ml / kg body weight at room temperature, the incision was 2-3 cm from the left abdomen; after local disinfection, the skin, subcutaneous layer, and muscular layer were cut layer by layer And peritoneum, found the left ureter, then sutured layer by layer. After local disinfection, check the marks and place them in corresponding mouse cages.
[0054] 2) Model control group: anesthetized and disinfected as above. The skin, subcutaneous...
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