Application of polypeptide modified nano particle to preparation of drugs for treating acute lung injury
A technology for acute lung injury and polypeptide modification, which is applied in the field of biomedicine to achieve the effects of facilitating in vivo tracking, promoting lung injury repair, and significant curative effect
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Embodiment 1
[0018] The preparation of embodiment 1 P12
[0019] The synthesis of gold nanoparticles is to adopt sodium citrate to reduce the chloroauric acid method, and the specific operation method is as follows: 6.45mL chloroauric acid (10.5mg / mL) is added into an Erlenmeyer flask filled with 200mL MiliQ water (18.2MΩ), and heated After heating to boiling on a stirring platform, 2 mL of sodium citrate (10%) was added quickly, and heating and stirring were continued for 15 minutes, then stirred and cooled to room temperature. Peptide (Pep 12: CLPFFD (SEQ ID NO: 1), purity>95%) powder was dissolved in water to prepare a peptide solution (1 mM). Mix the peptide and gold nanoparticle solution at a volume ratio of 1:10, and let stand overnight. Use a 220nm syringe filter head to filter the polypeptide gold nanoparticle suspension, then centrifuge (15000rpm, 30min), wash with PBS three times, and re-concentrate to 1mM for use.
[0020] The prepared polypeptide-modified nanoparticles are co...
Embodiment 2
[0021] Example 2 In vitro test proves that P12 can prevent neutrophils from migrating
[0022] 1. Experimental method
[0023] Take 20ml of human peripheral venous blood, inject it into a heparin anticoagulant tube, and shake it gently. Peripheral blood mononuclear cells were separated by Ficoll density gradient centrifugation. Take out the bottom layer containing erythrocytes, mix it with the same volume of 3% dextran, shake gently, let it settle naturally at room temperature for 1 hour, take the "supernatant part" after Dextran precipitation, centrifuge at 150g for 8 minutes, and resuspend in 0.2% Dextran In NaCl, add an equal volume of 1.6% NaCl after 1 minute, centrifuge at 150g for 8 minutes to remove red blood cells to obtain neutrophils, suspend them in PBS and count them, add CFSE (5 μM), and place the cells at 37 degrees for 30 minutes After centrifugation, wash twice with PBS containing 2% FBS, and suspend it in RPMI1640 medium (containing 10% fetal bovine serum, 5...
Embodiment 3
[0027] Example 3 In vivo experiments confirm that P12 can effectively treat acute lung injury
[0028] 1. Experimental method
[0029] A mouse model of acute lung injury was constructed. Select male, 6-8 week-old SPF grade C57BL6 mice, and weigh them. Give 5% chloral hydrate, inject intraperitoneally at a dose of 6 μl / g for anesthesia, and then fix it on the operating table in the supine position. Under sterile conditions, hold ophthalmic scissors to cut off the neck hair, and 75% medical alcohol cotton balls to disinfect the neck skin three times , the skin of the neck was cut about 0.5 cm in length, and the tissue was peeled off layer by layer with sterile ophthalmic forceps to expose the trachea. A 1ml syringe was selected to absorb sterile LPS (Sigma Company). The control group selected sterile PBS solution and underwent tracheal puncture. One hour before LPS instillation, instill 50 μL of P12 and P13 (1 μM) suspended in PBS. After the injection, the syringe was pulled...
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