Adenovirus Injection and Berberine Treatment

MiR-34b-5p and miR-470-5p overexpressing adenovirus vectors (Ad-miR-34b-5p and Ad-miR-470-5p) and their controls (Ad-miR-NC), BDNF overexpressing adenovirus vector or short guide RNA (sgRNA) adenovirus vector (Ad-BDNF and sg-BDNF) and their controls (Ad-NC and sg-NC) were constructed from Hanbio (Shanghai, China). CUMS mice were anesthetized by intraperitoneal injection of 10% chloral hydrate (4 mL/kg, Sigma-Aldrich, St. Louis, MO, USA) and fixed on a stereotactic instrument. A 0.5 mm diameter drill was used to open the skull on both sides of the mice. Subsequently, the adenovirus vectors (1011 pfu/mL, 1 µL) were slowly injected into the hippocampus using a glass microtube fixed on the stereotaxic instrument. After the operation, the mice were taken out of the stereotaxic instrument and placed in a warm cage to recover. On the third day after the operation, 20 mg/kg/day Berberine (dissolved in normal saline; Guangrui Bio, Shanghai, China) was administered by gavage. 2 weeks after the surgery, the mice were subjected to behavioral tests. In addition, the mice were euthanized and their hippocampus tissues were taken to detect the expression of miRNA and BDNF.

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