Objective: To study the effects of nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotide (ODN) on the preeclamptic umbilical serum induced expression of precollagen I, III mRNA and tumor necrosis factor-alpha (TNF-alpha) in cultured human umbilical artery smooth muscle cells (HUASMC).
Methods: Primary cultured HUASMC of normal pregnancy were divided into four groups: group A (HUASMC were incubated with umbilical serum of normal pregnancy); group B (HUASMC were incubated with umbilical serum of preeclampsia); group C (HUASMC were transfected with NF-kappaB cis decoy ODN 48 h before incubation with umbilical serum of preeclampsia); group D (HUASMC were transfected with NF-kappaB scramble ODN 24 h before incubation with umbilical serum of preeclampsia). NF-kappaB cis decoy ODN and NF-kappaB scramble ODN were transfected with cationic lipofectamine to the latter two groups, respectively. The proliferation of human umbilical artery smooth muscle cells was evaluated by methyl thiazolyl tetrazolium and the apoptosis was analyzed by flow cytometry. The expression levels of precollagen I, III mRNA were detected by RT-PCR, the expression levels of TNF-alpha were detected by western blot.
Results: (1) The proliferation of group B (0.19 +/- 0.02) and group D (0.18 +/- 0.03) was significantly increased as compared with those of group A (0.11 +/- 0.02) and group C (0.14 +/- 0.02) (P < 0.05). (2) The apoptosis rates of group B [(7.8 +/- 1.3)%], group C [(10.1 +/- 1.2)%] and group D [(8.1 +/- 1.3)%] were significantly reduced as compared with that of group A [(14.3 +/- 1.2)%] (P < 0.05), and there was a significant difference between groups B and C (P < 0.05). (3) The expression levels of precollagen I mRNA of group B (0.31 +/- 0.04), group C (0.23 +/- 0.04) and group D (0.30 +/- 0.03) were significantly increased as compared with that of group A (0.16 +/- 0.02) (P < 0.05), and there was a significant difference between groups B and C (P < 0.05). (4) There were no significant differences among the four groups in the expression level of precollagen III mRNA (P > 0.05). (5) The expression of TNF-alpha of group B (0.74 +/- 0.11), group C (0.36 +/- 0.09) and group D (0.79 +/- 0.12) were significantly higher than that of group A (0.15 +/- 0.03) (P < 0.05), and the expression of TNF-alpha of groups B and D were significantly higher than that of group C (P < 0.05), while there was no significant difference between groups B and D (P > 0.05).
Conclusions: NF-kappaB cis decoy ODN could down-regulate the proliferation, as well as the expression levels of precollagen and TNF-alpha of HUASMC induced by umbilical serum of preeclampsia. NF-kappaB may play an important role in the pathogenesis of placental artery abnormalities in preeclampsia.
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