Application of miRNA-Pa compound in preparing chronic pain diagnostic marker and therapy medicine
A technology for chronic pain, compounds for biomedical applications
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Embodiment 1
[0021] 1. Preparation of complete Freund's adjuvant (CFA) chronic arthritis pain model:
[0022] SPF grade male Kunming male mice of 6 weeks were selected under isoflurane anesthesia, and 10 μl of sterile complete Freund's adjuvant (complete Freund'sadjuvant, CFA; Sigma company) was injected intradermally at two points around the left hind foot metatarsal joint. This injection concentration can induce arthritis. 24 hours after injection, thermal and mechanical pain thresholds decreased and could be maintained for more than 3 weeks, and they were regarded as qualified mice for chronic pain modeling.
[0023] Pain behavior monitoring: Thermal hyperalgesia: Place the plexiglass box on a 3mm thick glass plate, and use thermal radiation stimulation according to the Hargreaves method to irradiate the mouse's sole close to the glass plate. Record the time from the beginning of irradiation to the appearance of the mouse's foot lifting, and the cut-off time is 20s. Each animal was me...
Embodiment 2
[0038] Example 2 miRNA-Pa treatment of chronic pain animal experiments
[0039] 1. Modeling and grouping:
[0040] SPF grade male Kunming male mice of 6 weeks were selected under isoflurane anesthesia, and 40 μl of sterile complete Freund's adjuvant was intradermally injected at two points around the left hind foot metatarsal joint. This injection concentration can induce arthritis. 24 hours after the injection, thermal and mechanical pain thresholds decreased and could be maintained for more than 3 weeks, as qualified mice for modeling; 20 mice in the miRNA-Pa intervention treatment group and 20 mice in the model control group (with phosphate buffer solution) PBS injection).
[0041] 2. Experimental method:
[0042] Intrathecal injection of miRNA-Pa intervention treatment group: After the mice were anesthetized with isoflurane, they were injected three times with miRNA-Pa-mimics analogs overexpressing miRNA-Pa (concentration 20 μM), 5 μl each time, along the spinal cord dur...
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