[Effect of microRNA-155 on inflammatory response and lipid uptake of macrophages and its mechanism].

Objective To investigate the effect of microRNA-155 on inflammatory response and lipid uptake of macrophages after the cells are stimulated by ox-LDL and its potential mechanism. Methods Macrophage RAW264.7 cells were treated with 0, 25, 50 and 100 μg/mL ox-LDL for 24 hours or with 50 μg/mL ox-LDL for 0, 6, 12, 24 hours. The level of miR-155 was evaluated in all above samples through real-time quantitative PCR. In our research, RAW264.7 cells were divided into six groups: control group, ox-LDL group, ox-LDL/negative control group, ox-LDL/anti-miR-155 group, ox-LDL/shRNA negative control group and ox-LDL/PPARγ-shRNA group. Oil red O staining was used to observe lipid uptake in the cells. Filipin staining was used to evaluate the cellular uptake of ox-LDL. Cholesterol testing was performed to examine the levels of total cholesterol (TC) and free cholesterol (FC). Real-time quantitative PCR was done to detect the expressions of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 mRNAs. According to study purpose, we explored the potential mechanisms of miR-155 inhibitor (including control group, ox-LDL group, ox-LDL/negative control group and ox-LDL/miR-155 inhibitor group), miR-155 mimic (including negative control group and miR-155 mimic group), and PPARγ shRNA (including control group, ox-LDL group, ox-LDL/shRNA negative control group and ox-LDL/PPARγ shRNA group) in ox-LDL-treated RAW264.7 cells through evaluating the expressions of p-STAT3, PPARγ, CD36 and NF-κBp65 using Western blotting. Results Ox-LDL stimulation increased the relative expression of miR-155 in a dose- and time-dependent manner. Through oil red O staining, Filipin staining, cholesterol testing and real-time PCR experiment, we found the relative absorbance, levels of TC and FC, filipin fluorescence intensity, and levels of TNF-α, IL-1β and IL-6 mRNAs were significantly lower in ox-LDL/anti-miR-155 group than in ox-LDL and ox-LDL/negative control group. Similarly, the relative absorbance, levels of TC and FC, filipin fluorescence intensity and levels of TNF-α, IL-1β and IL-6 mRNAs were significantly lower in ox-LDL/ PPARγ shRNA group than in ox-LDL group and ox-LDL/shRNA negative control group. The expressions of p-STAT3, PPARγ, CD36 and NF-κBp65 proteins were suppressed in ox-LDL/anti-miR-155 group as compared with ox-LDL group and ox-LDL/negative control group. Similarly, p-STAT3, PPARγ, CD36 and NF-κBp65 protein levels decreased in ox-LDL/PPARγ shRNA as compared with ox-LDL/vector group. Moreover, p-STAT3, PPARγ, CD36 and NF-κBp65 protein levels were higher in miR-155 mimic group than in negative control group. Conclusion Mediated by PPARγ, miR-155 induced inflammation response and lipid uptake of macrophages via STAT3/NF-κB signal pathway and CD36.