The concerted actions of microRNA-29a and interferon-β modulate complete Freund's adjuvant-induced inflammatory pain by regulating the expression of type 1 interferon receptor, interferon-stimulated gene 15, and p-extracellular signal-regulated kinase

Background

Previous research has shown that type 1 interferons (IFN), such as IFN-α and IFN-ß, possess antiviral and antinociception effects. Elevated levels of microRNA-29a (miR-29a) have been observed during inflammatory pain, and as miR-29a targets the type 1 IFN receptor (IFNR1), our study aimed to investigate the involvement of miR-29a, type 1 IFN, and IFNR1 in inflammatory pain.

Methods

Inflammatory pain was induced in male rats using complete Freund's adjuvant (CFA). The changes in miR-29a, IFN-ß, and IFNR1 were measured on Days 2, 3, 5, 7, and 10 post-CFA injection and expression of IFNR1, phospho-ERK (phosphorylated extracellular signal-regulated kinase) (p-ERK), extracellular signal-regulated kinase (ERK), and IFN-stimulated gene 15 (ISG15) were measured in rats that received an miR-29a inhibitor or miR-29a mimic.

Results

Our results demonstrated elevated miR-29a expression (CFA 3 days: mean difference [95% confidence interval, CI]: 0.860 [0.657–1.062]; CFA 5 days: mean difference [95% CI]: 1.120 [0.917–1.322], P<0.001, n=6) and decreased IFNR1 expression (CFA 3 days: mean difference [95% CI]: −0.300 [−0.470 to −0.130]; CFA 5 days: mean difference [95% CI]: −0.330 [−0.515 to −0.145], P=0.004, n=6) from Days 3–5 post-CFA induction, with IFN-ß expression showing a significant increase from Day 2 (F [3.30, 16.5]=34.3 for factor time, P≤0.01, n=6). Treatment with an miR-29a inhibitor alleviated CFA-induced mechanical allodynia and thermal hyperalgesia by Day 5 (P<0.001, n=9), concomitant with upregulation of IFNR1 and ISG15 expression, and downregulation of p-ERK (IFNR1; CFA 5 days + miR-29a inhibitor vs CFA 5 days; mean difference [95% CI]: 30.00 [20.31–39.69]; ISG15 conjugates; CFA 5 days + miR-29a inhibitor vs CFA 5 days, mean difference [95% CI]: 1.000 [0.9144–1.086]; free ISG15, mean difference [95% CI]: 2.402 [2.171–2.633]; p-ERK; CFA 5 days + miR-29a inhibitor vs CFA 5 days, mean difference [95% CI]: −32.00 [−34.10 to −29.90], P<0.001, n=9). Furthermore, in naïve rats, administration of an miR-29a mimic-induced mechanical allodynia, which was reversed by an ERK antagonist (P<0.001, n=6), associated with decreased IFNR1 and increased p-ERK expression (IFNR1; miR-29a mimic + dimethyl sulfoxide vs naïve; mean difference [95% CI]: −57.00 [−65.78 to −48.22]; miR-29a mimic + ASN007 vs naïve; mean difference [95% CI]: −60.00 [−71.00 to −49.00]. p-ERK; miR-29a mimic + dimethyl sulfoxide vs naïve, mean difference [95% CI]: 52.00 [47.01–56.99]; miR-29a mimic + ASN007 vs naïve, mean difference [95% CI]: 47.00 [42.51–51.49]; P<0.001, n=6).

Conclusions

Inhibiting miR-29a expression attenuates inflammatory pain by modulating IFNR1, ISG15, and p-ERK expression, highlighting the interactive roles of miR-29a and IFN-ß in the regulation of inflammatory pain.