| Objective: To investigate the effects of pravastatin on the expression of micro RNA-155(mi R-155)and the functions of 1ipopolysaccharide(LPS)-treated extravillous trophoblast cells.Methods: In vitro cultured HTR-8/SVneo cells were divided into the following groups:control group,enhanced plasmid with green fluoscent protein(p EGFP)-mi R-155 group(transfected with green fluorescent protein-tagged mi R-155),LPS group(treated with 100 ng / m L of LPS),mi R-155 inhibitor+LPS group,pravastatin+LPS group(treated with 100 ng/m L of LPS following pretreatment with 12.50,25.00,50.00 and 100.00 ?g/ml of pravastatin),and pravastatin+p EGFP-mi R-155 group(transfected with p EGFP-mi R-155 following pretreatment with 50 ?g/ml of pravastatin).Levels of mi R-155 in HTR-8 / SVneo cells treated with different strategies were measured by real-time polymerase chain reaction.Expression of phosphorylated Jun B(p-Jun B)and p-Fos B proteins was analyzed by Western blotting.Migration,invasion and apoptosis of HTR-8 / SVneo cells were also analyzed.All data were analyzed with t test.Results:(1)Compared with the control group,HTR-8/SVneo cells in the p EGFP-mi R-155 group were characterized by shorter migration distance [(274.70±18.82)vs(181.00 ±862)?m],less transmembrane cells[(123.00±4.36)vs(63.00±6.08)]and enhanced apoptosis[(5.40±0.68)%vs(9.27±0.68)%](all P<0.05).(2)Compared with the LPS group,the mi R-155 inhibitor+LPS group showed longer migration distance of HTR-8 / SVneo cells[(166.30±5.07)vs(242.00±18.07)?m,P<0.05],more transmembrane cells[(71.67±6.12)vs(109.00±7.81)P<0.05]and decreased cell apoptosis[(14.40±1.69)%vs(6.23±0.44)%,P<0.05].(3)Expression of mi R-155 at m RNA level in the LPS group was increased as compared with that ofthe control group(1.65±0.07 vs 0.79±0.12,P<0.05).Compared with the LPS group,pretreatment with 12.50,25.00,50.00 and 100.00 ?g/m1 of pravastatin decreased the expression of mi R-155 at m RNA level[(1.14±0.10),(1.02±0.10),(0.74±0.15)and(1.14+0.02)],especially at the concentration of 50 ?g/ml(all P<0.05).(4)Expression of p-Jun B and p-Fos B proteins in the control,LPS and pravastatin(50 ?g/m1)+LPS groups were(0.33±0.06)vs(0.37±0.07),(1.22±0.20)vs(0.80±0.13)and(0.31±0.02)vs(0.2l±0.05),respectively,showing higher expression level in both p-Jun B and p-Fos B proteins in the LPS group compared with that of the other two groups(all P<0.05).(5)Compared with the LPS group,the pravastatin(50 ?g/m1)+LPS group showed increased migration distance[(166.30±5.07)vs(246.80±13.42)?m,P<0.05],increased numbers of transmembrane cells[(71.67±6.12)vs(95.33±2.73),P<0.05]and decreased cell apoptosis[(14.40±1.69)vs(6.05±0.35)%,P<0.05].(6)Compared with the p EGFP-mi R-155 group,the pravastatin(50.00 ?g/ml)+p EGFP-mi R-155 group showed longer migration distance[(197.50±13.86)vs(275.80±13.63)?m,P<0.05],more transmembrane cells[(52.67±5.49)vs(125.00+6.66),P<0.05]and lower rate ofcell apoptosis[(8.90±1.00)vs(5.05±0.35)%,P<0.05].Conclusions: Pretreatment of extravillous trophoblast cells with pravastatin can protect them from apoptosis and loss of migratory and invasive abilities through inhibiting the activation of AP-1 and down-regulating the expression of mi R-155.which may be a mechanism that pravastatin inhibits the development of preeclampsia. |
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