Mycoplasma gallisepticum (MG) is the pathogen of chronic respiratory disease (CRD), hallmarked by vigorous inflammation in chickens, causing the poultry industry enormous losses. miRNAs have emerged as important regulators of animal diseases. Previous miRNA sequencing data has demonstrated that miR-130b-3p is up-regulated in MG-infected chicken embryo lungs. Therefore, we aimed to investigate the function of miR-130b-3p in MG infection of chickens. RT-qPCR results confirmed that miR-130b-3p was up-regulated both in MG-infected chicken embryo lungs and chicken embryonic fibroblast cells (DF-1 cells). Furthermore, functional studies showed that overexpression of miR-130b-3p promoted MG-infected DF-1 cell proliferation and cell cycle, whereas inhibition of miR-130b-3p weakened these cellular processes. Luciferase reporter assay combined with gene expression data supported that phosphatase and tensin homolog deleted on chromosome ten (PTEN) was a direct target of miR-130b-3p. Additionally, overexpression of miR-130b-3p resulted in up-regulations of phosphatidylinositol-3 kinase (PI3K), serine/threonine kinase (AKT), and nuclear factor-κB (NF-κB), whereas inhibition of miR-130b-3p led to the opposite results. Altogether, upon MG infection, up-regulation of miR-130b-3p activates the PI3K/AKT/NF-κB pathway, facilitates cell proliferation and cell cycle via down-regulating PTEN. This study helps to understand the mechanism of host response to MG infection.

1. Mycoplasma gallisepticum (MG) causes severe lung inflammation and cell damage by activating toll-like receptor (TLR) signalling, the nuclear factor-kappaB (NF-κB) pathway and pro-inflammatory cytokine gene expression. Baicalin (BA) is a flavonoid extracted from Scutellaria baicalensist, which possesses anti-inflammatory and anti-bacterial properties. This study investigated the effect of BA in MG-induced lung inflammation and its potential mechanism in MG-infected chicken embryo lungs and DF-1 cells. 2. The histopathological examination result showed that BA treatment alleviated MG-induced lung pathological changes. In addition, CCK-8 and cell cycle assays showed that BA treatment inhibited MG-induced cell proliferation and cell cycle progression in DF-1 cells. 3. The ELISA and RT-qPCR results demonstrated that BA treatment decreased the expression of interleukin-1beta (IL-1β), IL-6, and tumour necrosis factor-alpha (TNF-α) both in MG-infected chicken embryo lungs and DF-1 cells. 4. The results revealed that BA inhibited mRNA expression levels of toll-like receptor-6 (TLR6), myeloid differentiation primary response gene-88 (MyD88) and nuclear factor-κB (NF-κB), and the nuclear translocation of NF-κB-p65 5. In conclusion, the results showed that BA has a protective effect against MG-induced lung inflammation in chicken by inhibiting the TLR6-mediated NF-κB signalling.