A medicine for improving sperm motility and its pharmaceutical use
A technology of use and composition, which is applied in the field of pharmaceutical use of schisandrin A, and can solve problems such as undiscovered drugs
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Embodiment 1
[0026] Culture and processing of Leydig cells
[0027] 1 Culture method of Leydig cells
[0028] (1) Take the frozen TM3 cells out of the liquid nitrogen tank, put them into a water bath in a 37°C incubator immediately, and incubate the cryopreserved tubes for 2-3 minutes until they are completely thawed.
[0029] (2) Open the screw cap in the ultra-clean workbench, suck TM3 cells into a high-pressure steam-sterilized centrifuge tube, add 2ml DMEM / F12 high-glucose culture-based cryopreservation tube, wash the cells and transfer them to the centrifuge tube, and then add 2ml of DMEM / F12 high-glucose culture-based centrifuge tube, centrifuge at 800rpm for 5 minutes, discard the supernatant, add 3ml of medium, and repeatedly blow with a pipette to suspend the cells.
[0030] (3) Transfer the cell suspension to a disposable plastic culture bottle in the ultra-clean workbench again, then add about 2ml of medium, shake gently to spread the cells evenly on the bottom of the culture b...
Embodiment 2
[0045] Effects of Schizandrin A on the Vitality of Leydig Cells
[0046] The experiment was divided into normal group, DMSO group, and schisandrin A dosage groups. Schizandrin A dosage settings were as follows: 10 μmol / L, 20 μmol / L, 30 μmol / L, 40 μmol / L, 50 μmol / L, 60 μmol / L, 80 μmol / L. Schizandrin A was dissolved in DMSO.
[0047] After 24 hours of drug action, the supernatant was discarded, and the cells were washed 3 times with PBS. 10 μl of WST-8 solution was added to each well of the 96-well plate. After incubation in the incubator for 1 hour, the OD value was detected at a wavelength of 450 nm by a microplate reader.
[0048] statistical methods
[0049] SPSS 20.0 software was used to perform ONE-WAY ANOVA test on normally distributed data, and the mean value was the mean ± standard deviation Indicates that P<0.05 is a significant difference, which is statistically significant.
[0050] Experimental results
[0051] Compared with the normal group, 5‰DMSO could inhi...
Embodiment 3
[0053] Establishment of oligoasthenospermia model rats
[0054] The model rats were intraperitoneally injected with cyclophosphamide (Cyclophosphamide, CP) 35 mg / kg, once a day, for 5 consecutive days to create a rat model of oligoasthenospermia.
[0055]Grouping and administration of experimental animals: 70 SD rats were randomly divided into 7 groups, namely normal group 1, model group 1, Schizandrin A high, medium and low dose 3 groups, Wuzi Yanzong pill control group 1 and levocarni Ting control group 1 group, 10 rats in each group. Adaptive feeding for 3 days.
[0056] Normal group (NS): the same amount of CP was injected intraperitoneally with normal saline instead of CP for 5 days, and then intragastrically administered with normal saline from the 6th day. Continuous gavage for 2 weeks.
[0057] Model group (CP): intraperitoneal injection of CP 35 mg / kg for 5 days, and intragastric administration with normal saline from the 6th day. Continuous gavage for 2 weeks.
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