A skin repair polypeptide rl-rl10 and its application
A technology for skin repair and skin wounds, applied in the field of molecular biology, which can solve the problems of limited research reports
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Embodiment 1
[0020] Example 1: Animal model test of skin repairing polypeptide RL-RL10 promoting repairing activity.
[0021] Accurately weigh RL-RL10 dry powder and dissolve it in saline to prepare 25, 50, 100 nmol / L RL-RL10 polypeptide solutions. SPF grade Kunming mice weighing 22-25 g were selected as the test subjects, and the mice were randomly divided into 4 groups, with 3 mice in each group. Firstly, intraperitoneal injection of 1% pentobarbital sodium [dose: 1 µL / g (body weight)] was used to anesthetize the mice, and then the back hair of the mice was shaved and disinfected with 75% alcohol. Two symmetrical wounds of 8 mm×8 mm were drilled on both sides of the back with a punch. After the operation, the mice were placed next to the heater until they woke up and then returned to the feeding room to continue normal feeding. Three groups of mice were selected as treatment groups, and 20 μL of 1 mg / mL Fufuxin (KFX) was applied to the left wound of the treatment group mice, and 20 μL ...
Embodiment 2
[0023] Example 2: Detection of HaCaT cell proliferation activity of RL-RL10 polypeptide.
[0024] Human immortalized epidermal (HaCaT) cells were cultured in culture flasks. After the overgrowth, the HaCaT cells were digested with trypsin, centrifuged, the supernatant was discarded, and the cells were blown into a cell suspension with an empty medium (cell culture medium without fetal bovine serum), and then the cells were inoculated on 96-well plates (HaCaT: 5000 cells / well, 90 μL) were cultured for 2-4 h. After the cells adhered to the wall, the cells in the 96-well plate were divided into 4 groups, each with 3 wells, which were the control group, the 25 nmol / L RL-RL10 group, the 50 nmol / L RL-RL10 group, and the 100 nmol / L In the RL-RL10 group, physiological saline and RL-RL10 polypeptide solutions of different concentrations were placed in the corresponding wells. After culturing for 24 h, cells were detected using CellTiter 96® AQueous Single Solution Cell Proliferation ...
Embodiment 3
[0026] Example 3: Detection of HaCaT cell migration promoting activity of RL-RL10 polypeptide.
[0027] Migration of HaCaT cells was tested using 24-well culture plates and transwell chambers (8 μm wells; Corning, U.S.A.). HaCaT cells were cultured in culture flasks. When the bottom of the bottle is full, the digested cells are made into a cell density of 2×10 5 A single-cell suspension per mL, suspended in DMEM (serum-free), was added to each upper chamber (100 μL / well). Then, the cells were divided into 4 groups with 3 wells in each group, which were control group, 25 nmol / L RL-RL10 group, 50 nmol / LRL-RL10 group, 100 nmol / L RL-RL10 group, and normal saline Add different concentrations of RL-RL10 polypeptide solutions to the corresponding lower chamber (600 μL / well), and incubate at 37°C for 24h. On the upper surface of the chamber, non-migrated cells are carefully removed. The cells were fixed with methanol for 20 min, stained with 0.1% crystal violet for 20 min, and the...
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