Application of allicin and its derivatives in the preparation of medicines for preventing and treating kidney damage caused by urinary stones and crystals
A technology of urinary calculi and allicin, applied in the field of biomedicine, achieving good application prospects and the effect of inhibiting adhesion
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Embodiment 1
[0041] Example 1 A certain concentration of allicin DATS and DADS has no toxic effect on renal tubular epithelial cells NRK-52E and HK-2
[0042] 1. Materials and methods
[0043] 1. Main reagents
[0044] Dulbecco's Modified Eagle's Medium dry powder (DMEM, low sugar), fetal bovine serum (FBS), trypsin (typsin), dimethyl sulfoxide (dimethyl sulfoxide, DMSO), and Tris were purchased from Sigma-Aldrich (St.Louis, MO, USA); Allicin (Diallyl disulfide, DADS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Allicin (Diallyl trisulfide, DATS) was purchased from Guangdong Drug Control Institute. Other commonly used laboratory reagents are of domestic analytical grade.
[0045]2. Cell culture
[0046] At 37°C, 5% CO 2 And under the condition of saturated humidity, we used DMEM (low glucose) medium containing 10% fetal bovine serum to culture rat renal tubular epithelial cells NRK-52E and human renal tubular epithelial cells HK-2. The cell culture is carried out in a petri...
Embodiment 2
[0068] Example 2 Allicin and its derivatives have inhibitory effect on renal cell damage caused by sodium oxalate
[0069] 1. Experimental method
[0070] The experimental determination method of cell culture, MTT or MTS and the statistical method of other kits are the same as Example 1; 24 hours after the cells are seeded into the 96-well plate, different drug groups are set, and the different concentrations of the drug to be determined are added to the medium. Drug treatment for 24 hours. In the injury experiment caused by sodium oxalate (NaOx), allicin (DATS) was pre-added to the cells for treatment, and after 3 hours, soluble sodium oxalate of different concentrations was added to the cells (sodium oxalate combined with calcium ions in the medium Calcium oxalate crystals were formed later); 24 hours later, the corresponding MTS was added to measure the absorbance of each group, and the corresponding blank control wells were set (because calcium oxalate crystals and drugs ...
Embodiment 3
[0074] Example 3 Allicin and its derivatives have inhibitory effect on crystal adhesion after renal cell injury caused by sodium oxalate
[0075] 1. Experimental method
[0076] Materials are the same as Example 1.
[0077] Crystal adhesion experiment: First, cells were seeded into 6-well plates at a certain density (about 150,000-200,000 cells / well), and after 24 hours, the cells adhered to the wall. Change the medium and add different concentrations of allicin (DATS, DADS) and sodium oxalate (NaOx) to be measured for 24 hours. After aspirating and discarding the culture medium, add the prepared calcium oxalate suspension containing 20ug / cm2 per well, incubate at 37°C for 3 minutes, and then take out the cells. After the cells were washed 5 times with PBS, the cells were placed under a microscope to observe the cells, and the number of adhered crystals on the cell surface was counted under a field of view of 10*4. powerful.
[0078] All experiments were repeated at least ...
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