A miRNA-483-5p inhibitor drug and its application in medicine for treating osteoarthritis
A technology of osteoarthritis and inhibitors, applied in the field of biopharmaceuticals, can solve problems such as untreatable and preventable, and achieve the effect of delaying the progress of the disease
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Embodiment 1
[0028] The establishment of embodiment 1 osteoarthritis model
[0029] In this experiment, destabilization of the medial meniscus (DMM, destabilization of the medial meniscus) method was chosen as the experimental osteoarthritis mouse model. Wild-type male C57Bl / 6J and TG483 mice aged 8-10 weeks were selected for modeling. After the internal collateral ligament and anterior cruciate ligament were resected in the right knee joint of the mice in the experimental group, the internal meniscus was removed. In the sham operation group, only the skin incision of the right knee joint was performed. Typically, mice were sacrificed five weeks after surgery, and joint specimens were removed for subsequent testing.
Embodiment 2
[0030] Embodiment 2 intra-articular injection administration
[0031] A 33-gauge and a micro-syringe were used for intra-articular injection, and the virus titer injected in the administration group was 1×10 9 TU / ml, the concentration of antagomiR-483-5p (SEQ ID No.1) was 250 μM, the volume of each injection was 10 μl, and the injection was divided into two injections on the 7th day and the 14th day after operation.
Embodiment 3
[0032] The fixation of embodiment 3 tissue and the preparation of paraffin section
[0033] ①The complete right knee joint tissue was stripped and fixed in 4% paraformaldehyde fixative solution on a shaker at 4°C for 20-22 hours;
[0034] ② Rinse the fixed tissue with PBS for 5 minutes each time, twice in total. Then decalcification treatment is carried out, and the decalcification solution is 0.5MEDTA, pH 7.4. The decalcification solution was replaced every 2-3 days, and the total decalcification time was three weeks.
[0035] ③Automatic dehydrator for dehydration, transparency and wax dipping. Ethanol solution step by step dehydration, the specific program is: 50% ethanol solution, 2 hours; 70% ethanol solution, 2 hours; 80% ethanol solution, 2 hours; 95% ethanol solution, 2 hours; 100% ethanol 1, 1 hour; 100% ethanol II, 1 hour;
[0036]④ Transparent, that is, the ethanol solution is replaced by xylene. The specific procedure is: xylene I, 30 minutes; xylene II, 30 minu...
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