[Electroacupuncture improves pulmonary function by reducing inflammatory reaction via inhibiting miR-19b-3p to regulate SOCS3/JAK1/STAT3 signaling pathway in mice with chronic obstructive pulmonary disease]

Abstract

Objectives: To explore whether electroacupuncture(EA) can increase the expression of suppressor of cytokine signaling (SOCS)3 by affecting the expression of miR-19b-3p, inhibiting the continuous activation of janus kinase (JAK)1 /signal transducer and activator of transcription (STAT)3 signaling pathway, and improve pulmonary inflammation in chronic obstructive pulmonary disease (COPD) mice.

Methods: Forty mice were randomly divided into normal, COPD model, COPD+EA, and COPD+miR-19b-3p agomir (agomir)+EA groups. The COPD model was simulated by cigarette smoke exposure for 1 h, twice a day for 3 months. After modeling, mice in the COPD+EA group and the COPD+agomir+EA group received EA stimulation of "Feishu" (BL13) and "Zusanli"(ST36) for 30 min, once every other day, for 14 days. The mice of the COPD+agomir+EA group received intranasal drops of miR-19b-3p agomir solution (50 μL) 24 h before every EA intervention. The pulmonary ventilation functions including forced vital capacity (FVC), forced expiratory volume (FEV) at the 0.05 s and 0.1 s (FEV_0.05 and FEV_0.1), FEV_0.05/FVC and FEV_0.1/FVC were detected using a pulmonary function analysis system. The pathological morphology of lung tissue was observed after H.E. staining. The contents of interleukin(IL)-6, tumor necrosis factor(TNF)-α and IL-1β in the bronchoalveolar lavage fluid were assayed using ELISA. And the expressions of SOCS3, JAK1, STAT3, phosphorylated(p)-JAK1 and p-STAT3 proteins in the lung tissue were detected using Western blot. The expressions of miR-19b-3p, JAK1, STAT3 and SOCS3 mRNA were detected using real-time fluorescence quantitative PCR.

Results: Compared with the normal group, the COPD model group had a significant decrease in the levels of FVC, FEV_0.05, FEV_0.1, FEV_0.05/FVC and FEV_0.1/FVC, and the expression levels of SOCS3 mRNA and protein (P<0.001) and a significant increase in the contents of IL-6, TNF-α and IL-1β, and the expression levels of miR-19b-3p, JAK1 and STAT1 mRNAs, and p-JAK/JAK1 and p-STAT3/STAT3 protein expression ratio (P<0.001). After EA intervention, the decreased and increased levels of all the indexes mentioned above were reversed in the COPD+EA group (P<0.001, P<0.01, P<0.05). The effect of the EA+agomir were significantly less than EA in up-regulating the levels of FVC, FEV_0.05, FEV_0.1, FEV_0.05/FVC and FEV_0.1/FVC, and the expression levels of SOCS3 protein and in down-regulating the contents of IL-6, IL-1β, TNF-α, and expressions of JAK1 and STAT3 mRNA, and p-STAT3/STAT3 (P<0.001, P<0.01, P<0.05), suggesting that the effects of EA were weakened after intranasal drops of miR-19b-3p agomir. H.E. staining showed thickened alveolar wall, obvious inflammatory cell infiltration, with some ruptured alveoli fused into large vesicles in the model group, slightly dilated alveoli and small amount of inflammatory cell infiltration in the COPD+EA group, and slight alveolar fusion, slight thickening of alveolar wall, and light inflammatory cell infiltration in the COPD+agomir+EA group.

Conclusions: EA can inhibit the expression of miR-19b-3p, thereby up-regulating SOCS3 expression and inhibiting the overactivation of JAK1/STAT3 signaling, thus reducing lung inflammatory reaction to improve pulmonary function in mice with COPD.

Keywords: Chronic obstructive pulmonary disease; Electroacupuncture; JAK1/STAT3 signaling pathway; SOCS3; miR-19b-3p.