ABSTRACT
Matrix metalloproteinases 2 and 9 (MMP2/9) have previously been shown to be elevated in serum and amniotic fluid from women undergoing preterm birth. We performed experiments to determine the effects of MMP2/9 on uterine contraction and birth timing. Pregnant mice were injected daily with 50 mg/kg of SB-3CT or vehicle control beginning on gestational day 14-18 to determine if MMP2/9 inhibition would affect parturition timing. MMP2/9 expression in human myometrial tissue was determined by Simple Western (Wes) and semiquantitative western blot. Purified MMP2/9 and SB-3CT inhibitor were added to human myometrial strips to determine the effects of MMP2/9 on oxytocin-induced uterine contraction. Parturition was delayed in mice treated with MMP2/9 inhibitor SB-3CT. MMP2/9 protein levels were elevated in preterm laboring uterine myometrium. Gelatinase activity was confirmed in cell extracts and supernatants from immortalized and primary human uterine myometrial cells in culture. Addition of purified MMP2/9 increased the oxytocin-induced contractile response in myometrial tissue strips from pregnant women. In contrast, addition of the MMP2/9 inhibitor SB-3CT decreased the contractile response to oxytocin in a dose-dependent manner. These results suggest abnormal MMP2/9 expression affects the contractile state of the uterine myometrium to promote parturition and that MMP2/9 inhibition attenuates this effect.
Subject(s)
Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Myometrium/metabolism , Obstetric Labor, Premature/metabolism , Uterine Contraction/metabolism , Adult , Animals , Cells, Cultured , Disease Progression , Female , Heterocyclic Compounds, 1-Ring/pharmacology , Humans , Matrix Metalloproteinase 2/pharmacology , Matrix Metalloproteinase 9/pharmacology , Mice , Mice, Inbred C57BL , Myometrium/drug effects , Myometrium/pathology , Obstetric Labor, Premature/pathology , Oxytocin/pharmacology , Parturition/physiology , Pregnancy , Sulfones/pharmacology , Uterine Contraction/drug effects , Uterine Contraction/physiology , Uterus/drug effects , Uterus/metabolism , Uterus/pathology , Young AdultABSTRACT
Three new C-seco limonoids (1-3) and one new tetracyclic limonoid (4) were isolated from a methanol extract of the ripe fruits of Melia azedarach collected in Curitiba, Brazil, and their structures were elucidated by spectroscopic data analysis and comparison of spectral data with those of the previously known compounds. Among the limonoids isolated in the present study, compounds 3 and 4 exhibited significant inhibitory activity against HeLa S3 cancer cells, whereas 1 and 2 showed weak cytotoxicity.
Subject(s)
Limonins/chemistry , Limonins/pharmacology , Melia azedarach/chemistry , Brazil , Cell Proliferation/drug effects , Drug Screening Assays, Antitumor , Fruit/chemistry , HeLa Cells , Humans , Limonins/isolation & purification , Magnetic Resonance Spectroscopy/methods , Magnetic Resonance Spectroscopy/standards , Molecular Conformation , Reference StandardsABSTRACT
A methanol extract of the ripe fruits of Melia azedarach collected in Curitiba, Parana, Brazil, afforded seven new ring C-seco limonoids (1-7) together with three known limonoids (8-10). The structures of the new compounds were elucidated by NMR and MS analysis and comparison of spectral data with those of previously known compounds. Compounds 4 and 5 exhibited significant inhibitory activity against HeLa S3 cancer cells, whereas 1, 2, 3, and 8 showed weak cytotoxicity.
