The effect of curcumin on the expression of miR-155 to apoptosis and invasion of extravillus trophoblast cells treated by lipopolysaccharide

Abstract

Objective: To investigate the effect of curcumin on the expression of miR-155, apoptosis and invasion of extravillus trophoblast cells treated by lipopolysaccharide (LPS). Methods: Human trophoblast cells (HTR-8/SVneo cells) were divided into 4 groups, the curcumin + LPS group (pre-treated by curcumin of 12.5, 25, 50 μmol/L, then LPS of 100 ng/ml), the LPS group (100 ng/ml), the recombinant adenovirus group (miR-155, multiplicity of infection100) and the control group. The miR-155 level in HTR-8/SVneo cells was measured by real-time PCR, and the expression of NF-κB was analyzed by luciferase gene expression. The apoptosis of HTR-8/SVneo cells was tested by cell death detection ELISA and the level of NF-κB in HTR-8/SVneo cells was measured by western blot. In addition, transwell was used to test the invasive ability of HTR-8/SVneo cells in all the groups. Results: (1) The intracellular expression of miR-155 in the LPS group was (2.13±0.22) times of the control group (P<0.01); and the expressions of miR-155 in 12.5, 25, 50 μmol/L curcumin+LPS groups were (0.37±0.08) , (0.68±0.14) , (0.49±0.09) times as the LPS group, with statistically significant difference (P<0.05). (2) The expreesion of NF-κB in the LPS group was (2.25±0.56) times of the control group. The expreesion of NF-κB in the 12.5, 25, 50 μmol/L curcumin+LPS groups were (0.80±0.07) , (0.74±0.05) , (0.49±0.19) times to the LPS group, with statistically significant difference(P<0.01). (3)The p65 protein in the LPS group was (1.50±0.22) , significantly higher than the control group (0.95±0.25, P<0.01) . In 12.5, 25, 50 μmol/L curcumin+LPS groups, the p65 protein were (0.31±0.07) , (0.75±0.14) , (0.49±0.08) . Compared with the LPS group, p65 was down-regulated by curcumin, with statistically significant difference (P<0.05). (4) In the cell death detection ELISA, the A value in the control group, the LPS group and the recombined adenovirus group were (0.89±0.17) , (2.02±0.35) and (1.67±0.48) , respectively, with statistically significant difference (P<0.05). In the curcumin+LPS groups, when the curcumin concentrations were 25 or 50 μmol/L, the A value were (0.74±0.05) , (0.49±0.09) , respectively, compared with the LPS group(set as 1.00), with statistically significant difference (P<0.05). When the curcumin concentration was 12.5 μmol/L, the A value was (0.80±0.07) , with no statistical significance (P>0.05). (5) The transmembrane cells were (47±8), (60±14) in the LPS group and the recombined adenovirus group, respectively. Compared with the control group (169±18), the differences were statistically significant (P<0.05). In the curcumin + LPS groups, the transmembrane cells were (143±10), (137±10) when the curcumin concentrations were 12.5, 25 μmol/L, significantly higher than the LPS group (P<0.05) . The transmembrane cells were (55±7) when the curcumin concentration was 50 μmol/L, with no statistical difference(P>0.05). Conclusion: The treatment of curcumin could downregulate the expression of NF-κB/miR-155, thus inhibit NF-κB signal pathway and the apoptosis of extravillus trophoblast cells, and protect their invasive ability.