Application of mirna-9-5p compound in the preparation of chronic pain diagnostic markers and therapeutic drugs
A technology of drugs and pharmaceutical dosage forms, applied in the field of biomedicine
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Embodiment 1
[0020] 1. Preparation of complete Freund's adjuvant (CFA) chronic arthritis pain model:
[0021] SPF grade male Kunming male mice of 6 weeks were selected under isoflurane anesthesia, and 10 μl of sterile complete Freund's adjuvant (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.
[0022] 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 measured 3 times, and the ave...
Embodiment 2
[0036] Example 2 miRNA-9-5p inhibitor treatment of chronic pain animal experiments
[0037] 1. Modeling and grouping:
[0038] 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 injection, thermal and mechanical pain thresholds decreased and could be maintained for more than 3 weeks, and they were used as qualified mice for modeling; 20 mice were set up in the miRNA-9-5p intervention treatment group and 20 mice in the PBS-injected model control group .
[0039] 2. Experimental method:
[0040] Intrathecal injection of miRNA-9-5p inhibitor intervention treatment group in vivo: After mice were anesthetized with isoflurane, miRNA-9-5p antisense nucleotide (25 μM) (miR-9-inhibitor) was injected three times to down-regulate miRNA -9-5p expression, ...
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