Mir 223 agomir

Animal experiments were carried out in accordance with the guidelines approved by the Ethics Committee of Shenzhen University. The permit number of experiment animal for this study is SCXK (Yue) 2008-0002. Chronic hypoxia-induced PAH mouse model was performed as previously described55 (link). Lungs from PAH mice were stored at −80 °C prior to use. Rats for in vivo studies were randomized into four groups (n = 8 each): 1) a normoxic control group, 2) a hypoxic control group, 3) a hypoxic group injected with agomir control, and 4) a hypoxic group injected with miR-223 agomir (Ribobio). miR-223 agomirs (30 nmol), agomir control (30 nmol), or the normal saline (NS) were injected intravenously (tail vein, 0.3 ml) at day 7 and 14. Degree of PAH were assessed by measuring right ventricular systolic pressure (RVSP) and the ratio of right to left ventricle plus septum weight [RV/(LV + S)] at day 21 as described56 (link). After RVSP recording, whole blood were collected via right ventricular puncture, filled in 1.5 ml tubes, centrifuged for 3,000 × g for 10 min at 4 °C, and serum collected for detection of miR-223 expression levels.

BALB/C mice (8-week-old) were irradiated with a single dose of 800 cGy total body irradiation (TBI, Co60γ source). The aGvHD mice were infused with nucleated bone marrow cells (1 × 10⁷) and splenocytes (1 × 10⁷) isolated from C57BL/6j mice (6-week-old) through tail vein injection. After 24 h of irradiation, MSCs (5 × 10⁵/mouse, n = 15), chemically synthesized miR-223Agomir and micrON™ Ago NControl#22 (RiboBio, China) (10 nmol/mouse, n = 15) were used to treat aGvHD mice.
The miR-223Agomir or micrON™ Ago NControl#22, serving as the negative control, were centrifuged transiently to achieve miRNA powder aggregation at the bottom of the centrifugation tube. Subsequently, PBS buffer (1 ml) was added to dissolve the powder. The solution was injected into recipient mice via tail vein 1 day before irradiation, and 1, 4, and 7 days after irradiation.