Use of interferon-gamma as a cough drug target
A technology of interferon and drugs, applied in biological testing, biomaterial analysis, instruments, etc., can solve problems such as increasing cough sensitivity, and achieve the effect of increasing cough sensitivity and improving cough sensitivity
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Embodiment 1
[0070] Effects of IFN-γ pretreatment on cough sensitivity in whole guinea pigs
[0071] 1. Experimental animal grouping and administration method:
[0072]40 guinea pigs were randomly divided into 5 groups, and the guinea pigs were pretreated with different solutions: normal saline control group, osteopeptide control group (similar to high-dose IFN-γ group), low-dose, medium-dose and high-dose IFN - Gamma group (8×10 3 , 4×10 4 , 2×10 5 IU, using human IFN-γ).
[0073] The Buxco system was used to non-invasively measure the basic cough frequency of guinea pigs in an awake state (0.3M citric acid nebulized for 1 min, observed for 8 min).
[0074] The guinea pigs were anesthetized with ether, and under the guidance of the light source of the laryngoscope, 0.2 mL of normal saline containing the above-mentioned modeling agent was instilled into the trachea; twice a week for one week, and about 5 hours after the last infusion, the Buxco system was used to measure non-invasively...
Embodiment 2
[0078] Calcium Imaging Experiments
[0079] 1. Experimental method:
[0080] a. Primary vagus neuron culture:
[0081] Rats were anesthetized and bled to death, and bilateral vagus ganglia were obtained from the anterior neck craniotomy; they were carefully dissected, longitudinally torn, and chopped under a microscope; they were collected in a 5mL centrifuge tube containing cold PBS. Centrifuge at 400g×5min, remove the supernatant, add 0.5mL digestion solution, seal and digest in a water bath at 37°C for 40min. Add 2mL of cell inoculation solution, pipette carefully 40-60 times with a 1mL pipette tip (avoid air bubbles), and seal. Centrifuge at 400g×5min, remove the supernatant; add 2mL of cell inoculum, carefully pipette 30-40 times with a 1mL pipette tip (avoid air bubbles), and seal. Centrifuge at 400g×5min, remove the supernatant; add 2mL of cell inoculum, pipette carefully 20 times with a 1mL pipette tip (to avoid air bubbles), inoculate in a 35mm Petri dish containin...
Embodiment 3
[0090] patch clamp experiment
[0091] 1. Experimental method:
[0092] In this study, primary vagus neurons inoculated on 12mm round glass slides were used (the culture method was the same as in Example 2). First, the cell slides were taken out and washed with the electrode external solution for 3 times; the cell slides were pasted on the cell bath with high-vacuum sealing grease, and the electrode external solution was continuously perfused (the flow rate was 1 mL / min). Place the cell bath under an inverted microscope (Zeiss) connected to a CCD camera for testing.
[0093] Use the electrode pulling instrument to pull the glass electrode to the tip resistance of about 4 ~ 8MΩ (when the charging electrode is liquid in the tube). The chlorinated silver wire electrode needs to be immersed in the inner liquid of the electrode. Under the control of the micromanipulator, the tip of the glass electrode is sealed with the neuron cell membrane, and the resistance must reach GΩ or m...
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