Application of niclosamide in preparation of medicine for preventing and/or treating and relieving testis injury caused by Zika virus infection
A technology of niclosamide and Zika virus, applied in antiviral agents, drug combinations, diseases, etc., can solve the problem of no effective treatment drugs for the male reproductive system, protect male fertility, and reduce testicular spermatogenesis Effect of cell damage and relief of testicular symptoms
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Embodiment 1
[0039] Embodiment 1, animal infection experiment and treatment experiment
[0040] Male mice aged 6-8 weeks were infected by bilateral intraperitoneal route with 1 × 10 4 PFU ZIKV / mouse, the injection volume on each side was 50 μL (total volume 100 μL), the state of the mice was observed every day after infection, and the changes in body weight, disease incidence and survival rate of the mice were recorded. 5dpi after infection with ZIKV, the male mice in the niclosamide treatment group were intraperitoneally injected with 20mg / kg body weight of niclosamide every day until 14dpi, and the mice in the simple infection control group were injected with the same volume of PBS daily ; All mice were euthanized at 14dpi after infection. The whole blood of the mice was collected by removing the eyeballs, and the mice were killed by amputation. The main organs such as the brain, liver, lung, spleen, kidney, testis, and epididymis of the mice were collected, and the tissues were gently ...
Embodiment 2
[0041]Example 2. Inflammatory infiltration and seminiferous tubule injury of mouse testis treated with niclosamide
[0042] 1. Paraffin section
[0043] 1) Fixation: fix overnight in 4% PFA 10-15 times its own volume; the next day, remove the tissue from the fixative and place it in 75% ethanol overnight;
[0044] 2) Dehydration: After the fixed tissue is cut and trimmed, it is dehydrated in a dehydrator. The dehydration steps are: 70% ethanol, 3h; 80% ethanol, 3h; 90% ethanol I, 2h; 90% ethanol II, 2h; anhydrous Ethanol Ⅰ, 2h; Absolute ethanol Ⅱ, 2h;
[0045] 3) Transparency: After dehydration, the tissue pieces were soaked in xylene Ⅰ for 15 minutes; xylene Ⅱ for 15 minutes;
[0046] 4) Paraffin penetration: the transparent tissue is subjected to paraffin wax I, paraffin wax II, and paraffin wax III in sequence at a constant temperature of 52-56°C, 1 hour per step;
[0047] 5) Embedding: Soak the permeable tissue block in pre-melted paraffin, use a metal mold to make a pa...
Embodiment 3
[0061] Example 3, niclosamide inhibits the expression of S100A4
[0062] 1. Immunofluorescence staining
[0063] 1) Preparation of slices: ① Frozen section: dry the slices with an electric fan, fix them with cold acetone (place the acetone at 4°C overnight for pre-cooling) in a refrigerator at -20°C for 10 minutes, blow the slices with their backs facing the electric fan for 1 hour, and place them in - Store at 20°C for later use; ②paraffin sections: preheat paraffin sections at 55-60°C for 2 hours, and then routinely dewax: immerse the sections in xylene Ⅰ for 15 minutes; xylene Ⅱ for 15 minutes; absolute ethanol Ⅰ for 10 minutes; Ethanol Ⅱ, 5min; 90% ethanol 5min; 70% ethanol 5min;
[0064] 2) Washing: washing with ddH2O, 5 min×1 time, washing with PBS, 5 min×3 times;
[0065] 3) Sealing: Incubate slices with 5% BSA-PBS at room temperature for 1 hour; no need to protect from light, but place in a wet box to prevent dry slices;
[0066] 4) Primary antibody: Discard the blo...
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