Application of cordycepin in preparation of medicine for preventing and/or treating radiation damage
A technology of cordycepin and drug, applied in the field of medicine, can solve the problems such as no relevant literature reports on the effect of cordycepin, unclear effect of cordycepin, etc., to enhance antioxidant capacity and colony formation capacity, promote the recovery of hematopoietic system, and reduce cell counts. The effect of vitality
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Embodiment 1
[0036] Detection of Antioxidant Proteins
[0037] Dissolve cordycepin in low-sugar medium, so that the concentration of cordycepin in low-sugar medium is 200 μM. Human dermal fibroblasts were cultured in low-sugar medium containing 200 μM cordycepin for 0, 2, 4, 6, 8, 12, 24, 48, 72, and 96 h, respectively. After the cells were cultured in low-glucose medium containing cordycepin, the cells were collected and protein lysate was added to lyse the cells. The protein concentration was detected by the BCA method. The denatured protein was electrophoresed, transferred to a membrane, and blocked. The primary antibody was blocked overnight and then developed with a secondary antibody. The detected antioxidant proteins include SOD1, SOD2, and GPX-1. The results showed that the levels of antioxidant proteins (SOD1, SOD2, GPX-1) increased in cells pre-cultured with cordycepin ( figure 1 a).
[0038] colony formation assay
[0039] The cordycepin was dissolved in the low-sugar medium, ...
Embodiment 2
[0041] γ-H2AX formation assay
[0042] Dissolve cordycepin in low-sugar medium, so that the concentration of cordycepin in low-sugar medium is 200 μM. Human dermal fibroblasts were cultured in low-sugar medium containing 200 μM cordycepin for 72 hours. After 72 hours, the cells were seeded on coverslips, cultured (low-sugar medium without cordycepin) overnight and irradiated with 5Gy gamma rays. Cells were fixed with 4% paraformaldehyde at different times (0.5, 2, 6, 12, 24 h) after irradiation, cultured in 1% triton solution, blocked with goat serum for 30 minutes, incubated overnight at 4°C with γ-H2AX antibody, and goat anti-mouse The fluorescent secondary antibody was incubated at 37°C in the dark for 2 hours, and after counterstaining the nuclei with DAPI, images were collected with a fluorescence microscope to observe the formation of γ-H2AX. The results showed that, compared with the control cells, cultured with cordycepin could significantly reduce the formation of γ-...
Embodiment 3
[0047] Human dermal fibroblasts were seeded in 96-well plates (1×10 4 / well) (low-sugar medium without cordycepin) overnight. Then, 0, 100, 200, and 300 μM concentrations of cordycepin were added to the cells at 37°C, 5% CO 2 Conditioned for 72h. with 900 μM H 2 o 2 After culturing for 12 hours, the relative cell viability was detected by CCK-8 assay. The results showed that compared with the control cells, the cells cultured with cordycepin could significantly reduce the H 2 o 2 Cell viability decreased due to culture ( image 3 ).
[0048] Seed human bronchial epithelioid cells (HBE), tumor A549 and H460 cells in 96-well plates
[0049] (1×104 / well) medium culture (low-sugar medium without cordycepin) overnight. Then, cordycepin at concentrations of 0, 50, 100, 200, and 300 μM was added to the cells and cultured at 37° C. and 5% CO 2 for 72 hours. After 72 h, the relative cell viability was detected by CCK-8 assay. The results showed...
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