miR‐495 targets ROCK1 to inhibit lipopolysaccharides‐induced WI‐38 cells apoptosis and inflammation

Abstract

Abstract Pneumonia is an inflammatory disease with leading mortality rate in children. It has been well established that microRNAs (miRNAs) have been regarded as critical regulator in acute lung injury. We intended to explore the effect and underlying mechanism of miR‐495 on lipopolysaccharides (LPS)‐induced WI‐38 cells. Here, we first found that miR‐495 was downregulated in serum of patients with acute stage pneumonia. To establish cell model of acute pneumonia, WI‐38 cells were treated with 20 μg/mL LPS, and qRT‐PCR analysis also confirmed the downregulation of miR‐495 in LPS‐induced WI‐38 cells. Data from MTT (3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide) and flow cytometry assays showed that the decreased cell viability and induced cell apoptosis by LPS treatment were also reversed by miR‐495 over‐expression. Moreover, miR‐495 inhibited expression of associated inflammatory factors, which were induced by LPS treatment. Second, ROCK1 (rho‐associated, coiled‐coil‐containing protein kinase 1) was identified as functional target gene of miR‐495, whose expression was decreased by miR‐495. Mechanically, combination of miR‐495 and ROCK1 over‐expression reversed the influence of miR‐495 on LPS‐induced inflammation, viability, and apoptosis. In conclusion, our findings indicated that miR‐495 inhibited LPS‐induced inflammation injury and apoptosis in WI‐38 cells via targeting ROCK1, which would shed light on therapeutic schedule in acute pneumonia.

Keywords

• acute pneumonia
• apoptosis
• inflammation
• lipopolysaccharides
• miR‐495
• ROCK1