Objectives
Uterine contractility may be initiated through inflammation-related pathways. Progesterone, an endogenous hormone, is thought to mitigate preterm labor partly through anti-inflammatory effects. Our lab has previously identified mono-hydroxylated metabolites of progesterone with differential activities related to uterine contractility. Our objective was to investigate the anti-inflammatory properties of progesterone and major progesterone metabolites on uterine smooth muscle cells.
Methods
Primary uterine smooth muscle cells (UtSMC) plated at 2 x 10 ∧ 5 in a 12-well dish using smooth muscle growth media (Lonza) achieved approximately 60-70% confluence after allowing 24 hr for cell adherence. UtSMC cells were then treated with progesterone (P4) and major progesterone metabolites (17α-hydroxyprogesterone, 16α-hydroxyprogesterone, 6β-hydroxyprogesterone) at 1μM for 24 hr. UtSMC were subsequently exposed to LPS (1μg/mL) for 24 hr in addition to replacement of their progestin treatment. UtSMC treated with DMSO with and without LPS exposure served as negative and positive controls, respectively. Culture media was collected and IL-6 concentrations measured by ELISA. GraphPad Prism software was used for statistical analysis (t-test). Results are expressed as mean +/- SEM and p<0.05 was considered significant.
Methods
Primary uterine smooth muscle cells (UtSMC) plated at 2 x 10 ∧ 5 in a 12-well dish using smooth muscle growth media (Lonza) achieved approximately 60-70% confluence after allowing 24 hr for cell adherence. UtSMC cells were then treated with progesterone (P4) and major progesterone metabolites (17α-hydroxyprogesterone, 16α-hydroxyprogesterone, 6β-hydroxyprogesterone) at 1μM for 24 hr. UtSMC were subsequently exposed to LPS (1μg/mL) for 24 hr in addition to replacement of their progestin treatment. UtSMC treated with DMSO with and without LPS exposure served as negative and positive controls, respectively. Culture media was collected and IL-6 concentrations measured by ELISA. GraphPad Prism software was used for statistical analysis (t-test). Results are expressed as mean +/- SEM and p<0.05 was considered significant.
Stay updated, free articles. Join our Telegram channel
Full access? Get Clinical Tree
