Middle cerebral artery occlusion
Rats underwent surgery for right middle cerebral artery occlusion (MCAO). After anesthetisation, a nylon filament with a silicon tip of a diameter of 0.37 mm was inserted into the right MCA to obstruct blood flow for 2 h. Subsequently, it was removed to allow reperfusion. Regional cerebral blood flow was observed using transcranial laser Doppler to confirm a decrease in blood flow by 20%–30% compared with the baseline value during MCAO. Sham-operated rats underwent the same anaesthesia surgical procedures without MCAO. After recovering from anaesthesia, the rats were returned to sterilised cages with regular access to food and water.
Rats were randomly divided into six groups (n = 12/group): sham group; MCAO + control group; MCAO + miR-29b antagomir group; MCAO + H19 siRNA group; MCAO + miR-29b antagomir group + H19 siRNA control group; MCAO + miR-29b antagomir + H19 siRNA group. miR-29b antagomir, H19 siRNA, and transfection reagent were purchased from GenePharma (Shanghai, China). Three days before MCAO, the rats were treated using a mixture of 100 μL miR‐29b antagomir, H19 siRNA, or control + 32‐μl transfection reagent through the tail vein. The sequences were 5’-CCACCGUAAUUCAUUUAGATT-3’ (H19 siRNA), 5’‐AACACUGAUUUCAAAUGGUGCUA-3’ (miR-29b antagomir), and 5’-UUCUCC GAACGUGUCACGUTT-3’ (H19 siRNA control).