Abstract
BackgroundA recent study found that Salvia miltiorrhiza extract substantially increased amniotic fluid index in women with oligohydramnios. Abnormal aquaporin (AQP) expression is associated with oligohydramnios. This study aims to investigate the effect of tanshinone IIA, the major lipophilic component in the root of Salvia miltiorrhiza, on the function and expression of AQPs in human amniotic epithelial WISH cells and the molecular mechanism underlying the effect. MethodsWISH cells were treated with tanshinone IIA. Cell proliferation and swelling were determined by cell counting kit-8 assay and measurement of cell size, respectively. A human phospho-MAPK antibody array was used to screen tanshinone IIA-associated signaling molecules. The protein expression of AQPs, glycogen synthase kinase 3β (GSK-3β), and phosphorylated GSK-3β (p-GSK-3β) was analyzed by Western blot. ResultTanshinone IIA significantly inhibited WISH cell proliferation at ≥50μM (P<0.05) and the IC50 was 35μM. Cell size was significantly increased by the treatment with 35μM tanshinone IIA for 24h (P<0.05). WISH cells expressed AQP1, AQP3, AQP8, and AQP9 predominantly in cytoplasm. The dose of tanshinone IIA to maximally induce AQP1 and AQP3 expression was 35μM; for AQP8 and AQP9, the dose of maximal stimulation was 5μM (All P<0.05). Tanshinone IIA (35μM)-mediated up-regulation of AQP8, AQP9, AQP1, and AQP3 peaked at 6, 12, 24, and 24hours of treatment, respectively. Tanshinone IIA increased p-GSK-3β level significantly (P<0.05). Blockade of GSK-3β expression by siRNA and inhibition of GSK-3β activity by lithium chloride significantly reduced tanshinone IIA-induced AQP1 and AQP3 expression (All P<0.05). ConclusionTanshinone IIA increases AQPs expression in WISH cells by stimulating p-GSK-3β. AQP8 responds to tanshinone IIA the most rapidly compared with AQP9, AQP1, and AQP3 in WISH cells.
Published Version
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call