Induction of CIA and miR-143-3p treatment
The CIA mice model was prepared as previously
described[19,20]. DBA/1J mice were used for CIA
induction by bovine type II collagen (CII) Chondrex, Inc., Washington,
DC, USA), acetic acid, and Freund’s adjuvant (CFA) (Sigma-Aldrich, St.
Louis, MO, USA). The day of the first immunization was determined on day
0. On day 21, 0.1 mL CII emulsified with incomplete Freund’s adjuvant
(IFA) (final concentration of 1 mg/mL) was i.d. in the tail, proximally
to the primary injection site, to enhance immunization.
A total of 24 DBA/1J mice were randomly divided into 4 groups as follows
(n = 6 per group): control group, normal mice were injected with
saline (1 mL/kg, i.v.); model group, CIA mice administered with saline
(1 mL/kg, i.v.); control mimics group, CIA mice were injected
intravenously with a 200 μL mixture of EntransterTM-in vivo (25 μL,
Engreen Biosystem Co, Ltd., Beijing, P.R. China) and control mimics (50
μg); miR-143-3p mimics group, CIA mice were injected with a 200 μL
mixture of EntransterTM-in vivo (25 μL) and miR-26b-5p mimics (50 μg).
MiR-143-3p mimics or the control substances were administered at day 28
and day 35 after the first immunization[21]. The
paw thickness and arthritis severity were detected at day 22, 26, 30,
34, 38, 42. Mice were sacrificed on day 44, and spleens, lymph nodes,
plasma, and keen joints were obtained for further studies.