Pharmaceutical composition for preventing or treating hearing loss
A technology for hearing disorders and cilostazol, which is applied in the field of pharmaceutical compositions for preventing or treating hearing disorders, and achieves the effects of treating noise-induced hearing disorders or ototoxic hearing disorders and inhibiting apoptosis
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Embodiment 1
[0053] Embodiment 1: According to the effect analysis of compound preparation of the present invention on hearing function and vestibular function
[0054] Preparation of compound preparation
[0055] Cilostazol (CS, SK Chemicals) and Ginkgo biloba extract (GbE, SK Chemicals) were mixed in a weight ratio of 5:4 and dissolved in 0.5% (w / v) carboxymethylcellulose sodium (CMCNa, Sigma) in an aqueous solution to prepare a composite preparation (hereinafter referred to as renexin) at a concentration of 18 mg / mL.
[0056] Application of sample
[0057] 52 SD (Sprague-Dawley) rats (200-250 g) aged 8-10 weeks were divided into the following 6 groups, and each drug was administered as follows.
[0058] -The first group: normal control group (n=3);
[0059] -The second group: cisplatin (CDDP) administration group alone (n=13);
[0060] -The third group: ranashin+CDDP administration group (n=12);
[0061] - The fourth group: Ginkgo biloba extract+CDDP administration group (n=12); ...
Embodiment 2
[0078] Example 2: Morphological evaluation of auditory and vestibular hair cells administered according to the compound formulation of the present invention
[0079] Morphological evaluation of the middle and bottom
[0080] The rats in each group of Example were anesthetized, and then their temporal bones were removed. Under a dissecting microscope, in the cochlea The round window (round window) and oval window (oval window) were drilled, and the perilymphatic space (perilymphatic space) was perfused with 4% glutaraldehyde (EMgrade, place of origin, Inc.CA, USA) with a Pasteur pipette. Each sample was then fixed in 4% glutaraldehyde solution for 24 hours. The surface of the cochlea and the perilymphatic space were washed three times with phosphate buffered solution (PBS), perfused with 1% osmium tetroxide, and placed on a tissue rotator for 15 minutes. Then, the sample was washed 3 times with PBS, the bony capsule of the cochlea was removed under a dissecting microscope...
Embodiment 3
[0092] Embodiment 3: Analysis of the in vitro effect of the compound preparation of the present invention on CDDP-induced cytotoxicity
[0093] Confirmation of CDDP-induced cytotoxicity
[0094] In order to confirm CDDP-induced cytotoxicity, the auditory cell line (HEI-OC1 (ATCC, USA) was cultured at 33°C with 10% CO2 in Dulbecco's essential medium (DMEM, Dulbecco's minimum essential medium) was cultured. The cultured cells were divided into 96-well plates at a concentration of 3000 cells / well, and then 20 μM cisplatin was added for culture. When cultivating the first 0, 12, 24 and 48 hours, 20 μl of MTS solution was added to each well, and the culture was continued for 4 hours, and then the absorbance of formazan was measured at 490 nm by a fully automatic quantitative drawing microplate reader, so as to determine the proportion of surviving cells.
[0095] The measurement results are shown in Figure 8 . Such as Figure 8 As shown, according to the treatment with cisplati...
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