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6-shogaol (bioactive metabolite of Zingiber Officinale) attenuates human uterine smooth muscle (USM) contractility
Abstract Number: GM-2
Abstract Type: Original Research
Objective: Pre-term labor is a major health care challenge in need of novel and more effective tocolytics. 6-shogaol (6S) is a purified component of Zingiber Officinale (ginger) which has been shown to relax airway smooth muscle and inhibit GPCR q-coupled stimulation of intracellular calcium levels [Ca2+]i (1),(2). Given the central role GPCR q signaling plays in myometrial contractility (3), we questioned if 6S treatment would relax pre-contracting pregnant human USM ex vivo and whether 6S could attenuate oxytocin-induced [Ca2+]i release in vitro.
Ex vivo organ bath experiments were performed utilizing strips of late gestation human USM (n=5 patients). Samples were pre-contracted with oxytocin (OXY; 0.5 μM), equilibrated for 60 minutes, and treated with sequentially increasing doses of 6S (1 - 500 µM) or vehicle control (0.1% DMSO final) to determine IC50, Imax, and minimum effective dose (Figure 1A). Changes in force were measured over time and calculated as a percent reduction in integral force (g*sec) from baseline oxytocin-induced contractility.
To study the effect on [Ca2+]i, immortalized human USM cells were loaded with the Ca2+ specific fluorophore Fura-2 AM (5 µM), pretreated (10 min) with vehicle (0.1% DMSO) or sequentially increasing doses of 6S (1 - 500 μM), followed by stimulation with OXY(1 µM). Calcium fluorescence (340/380) was recorded in the FlexStation 3 plate reader. Results were compiled and reported as mean ± SEM. Data was analyzed by one way ANOVA with Bonferroni's Multiple Comparison Test, and p<0.05 was taken as significant.
The IC50, Imax and minimum effective dose of 6S on OXY- induced contractility of human USM are 43 μM,250 μM and 10 μM (***p<0.001), respectively (Figure 1B). The IC50, Imax and minimum effective dose of 6S on OXY- induced intracellular calcium response in human USM cells are 30 μM,100 μM and 10 μM (***p<0.001), respectively (Figure 1C).
Conclusions: 6S relaxes pre-contracting pregnant human USM tissue and reduces oxytocin-induced elevations in calcium in human USM cells. These findings may lead to the development of a novel class of drugs for treatment of preterm labor.
(1) Townsend EA, Zhang Y, Xu C, Wakita R, Emala CW. Am J Resp Cell Mol 2014, 50(1):115-124.
(2) Townsend EA, Siviski ME, Zhang Y, Xu C, Hoonjan B, Emala CW. Am J Resp Cell Mol 2013, 48(2):157-163
(3) Andrés López Bernal. BMC Pregnancy Childbirth. 2007, 7(Suppl 1): S2.