Use of nerve growth factor in preparing medicine for treating middle-aged and elderly male hypogonadism syndrome
A nerve growth factor, middle-aged and elderly technology, applied in the field of biomedicine
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Embodiment 1
[0035] Example 1: Detection of the pro-proliferation effect of rat nerve growth factor on rat Leydig cells
[0036] Materials: Coming 12-well plate (purchased from Corning, USA, product number: 3336), testosterone radioimmunoassay kit (purchased from Beijing North Institute of Biotechnology, product number: B10TFB), human nerve growth factor (purchased from Sigma-Aldrich Shanghai Trading Co., Ltd., product number: N1408-.1MG), rat nerve growth factor (purchased from Sigma-Aldrich Shanghai Trading Co., Ltd., product number: N2513-1MG) or mouse nerve growth factor (purchased from Sigma-Aldrich Shanghai Trading Co., Ltd. Ltd., product number: SRP4304-20UG), insulin-transferrin-sodium selenite medium supplement (ITS, purchased from Sigma-Aldrich Shanghai Trading Co., Ltd., product number: I1884), luteinizing hormone (LH, purchased from From Sigma-Aldrich Shanghai Trading Co., Ltd., item number: L9773), ethanedimethanylsulfone (EDS, synthesized by the School of Chemical Engineering...
Embodiment 2
[0039] Example 2: Detection of the differentiation-promoting effect of rat nerve growth factor on rat testicular mesenchymal stem cells
[0040] Male SPRAGUE-DAWLEY rats (purchased from Guangdong Provincial Medical Animal Experiment Center, 12 weeks old, 250 ± 20 g / rat) were intraperitoneally injected with ethanedimethylsulfone (EDS, 90 mg / kg body weight) 7 days before the experiment, and carbon dioxide After execution, the testes were taken out, placed in ice-cold phosphate buffer, the capsule was cut off, the blood vessels and seminiferous tubules were stripped, and the seminiferous tubules were separated into single ones, and placed in DMEM / F-12 medium (containing 0.1% Bovine serum albumin and 1x insulin-transferrin-sodium selenite medium supplement) were cultured overnight at 34°C and 5% carbon dioxide. The next day, the seminiferous tubules were divided into 24-well plates, and rat nerve growth factor was added for treatment. The concentration of rat nerve growth factor w...
Embodiment 3
[0042] Embodiment 3: Radioimmunoassay analysis of testosterone content in culture medium
[0043]Take 90-day-old SAMR1 mice (3 months old, body weight 26±2g), use forceps to extract the seminiferous tubules in the testes under sterile conditions, wash them with phosphate buffer for 6 times, and then culture them with DMEM / F-12 The base was washed 3 times, and the final concentration of 375 micrograms / milliliter (ug / ml) was added to the DMEM / F-12 medium containing 0.1% bovine serum albumin, and 24 Hour. After washing 3 times, change the fresh medium, then place it in an environment of 34°C and 5% carbon dioxide to cultivate, and change it every 3.5 days to contain different concentrations (10, 100 and 200 ng / ml) of human nerve growth factor, Cell culture medium for luteinizing hormone (LH) and platelet-derived growth factor AA (PDGFAA). Finally, the testosterone content in the medium was analyzed by radioimmunoassay. The experimental results showed that on day 14, nerve grow...
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