Application of curcumin in preparation of medicine for preventing and treating ionizing radiation induced reproductive system injury
A technology of ionizing radiation and reproductive system, applied in the application field of curcumin and medicine, can solve problems such as unreported, and achieve the effect of improving apoptosis, wide source and good development prospect.
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Embodiment 1
[0033] Example 1 – Cytotoxicity detection of curcumin on mouse spermatocytes
[0034] The present embodiment selects mouse spermblast GC-2, which is more recognized for reproductive system injury in vitro studies. Curcumin was dissolved in dimethyl sulfoxide with a mother liquor concentration of 20 mmol / L, and curcumin solutions with concentrations of 4, 8, 12, 16 and 20 μmol / L were prepared by the method of step-by-step dilution, and the dilution was DMEM culture solution for culture cells. The control group was treated with dimethyl sulfoxide in equal volumes of 20 μmol / L periloxal solution. GC-2 cells are seeded in 96-well plates, with the addition of the above different concentrations of curcumin solution and dimethyl sulfoxide when the cells are in the logarithmic growth phase, respectively, and cell growth is detected using CCCK8 (Dojindo, the operation method follows the instructions) after 48 h of treatment.
[0035] The test results of the above experiments are as follows ...
Embodiment 2
[0036] Example 2 – Protective effect of curcumin on decreased viability of mouse spermatocytes caused by ionizing radiation
[0037] In the present embodiment, GC-2 cells (1×10 are taken 5 Inoculate in 96-well culture plates, 100 μL / well, 100 μL of drug per well (curcumin solution), so that the final concentration is 12, 8, 4 μmol / L, 5 wells per concentration, and a blank control group cell suspension and irradiation group cell suspension are also provided. 4Gy irradiation was performed 2h after the drug action, and CCK8 was used to detect cell growth 24h after the photo.
[0038] The test results of the above experiments are as follows Figure 2 As shown, curcumin increased spermocyte viability by more than 10% at 4-12 μmol / L to γ-ray 4Gy, suggesting that curcumin pretreatment played a protective role in the decrease in spermatocyte viability caused by ionizing radiation.
Embodiment 3
[0039] Example 3 – Protective effect of curcumin on apoptosis of mouse spermatocytes caused by ionizing radiation
[0040]In this embodiment, GC-2 cells were seeded in a 6-well culture plate, and four groups were established: (1) no treatment group; (2) 10 μmol / L curcumin group; (3) 4Gy ionizing radiation group; (4) 4Gy ionizing radiation + 10 μmol / L curcumin group. Corresponding to the experimental conditions after treating the cells 24 h, centrifuge tubes to collect cell culture solution and pancreatic enzyme digestion of the cells, 2000 × g centrifugation for 5 min, discard the supernatant, resuspend the cells with PBS and count. Take 5-100,000 resuspended cells, centrifuge 1,000 × g for 5 min, discard the supernatant, add 195 μL of Annexin V-FITC binding solution and gently resuspend the cells. Add 5 μL annexin V-FITC and mix gently. Add 10 μL propidium iodide staining solution and mix gently. Incubate at room temperature for 10 min with flow cytometry.
[0041] The test resul...
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