) at a wavelength of A550 nm. The amount of FC absorbed into the H. pylori cells was quantified

based on the FC-standard curve and calculated per the CFU. Helicobacter pylori cell suspension (600 μL) was cultured for 24 h with various volumes of FC beads (FC concentration: 30–90 μM) in a simple-PPLO broth (15 mL) containing progesterone (30 μM), with continuous shaking under microaerobic conditions in the dark, and the CFUs were Selleck BTK inhibitor then measured. Next, an H. pylori cell suspension (600 μL) was cultured for 24 h with various concentrations of progesterone (from 10 to 30 μM) in a simple-PPLO broth (15 mL) containing FC beads (FC concentration: 500 μM) or the FC-free beads (in a similar volume), with continuous shaking under microaerobic conditions in the dark, and the CFUs were then measured. In our first experiments, we investigated the effects of the steroid hormones estradiol, androstenedione, and progesterone in inhibiting the growth of H. pylori. NSC 683864 mouse When H. pylori (approximately 105.5 CFU mL−1) was cultured for 24 h in different simple-PPLO broths (3 mL) containing single steroids at concentrations ranging from 10 to 100 μM, every steroid hormone examined exhibited inhibitory effects on the growth of H. pylori at concentrations >50 μM (Fig.

1). Estradiol appeared to act bacteriostatically on H. pylori, as the CFUs of H. pylori cultured in the presence of estradiol at the 50 and 100 μM concentrations were entirely unaltered from the baseline CFU (105.5 CFU mL−1) before the cultures (Fig. 1a). In contrast, androstenedione and progesterone Thymidylate synthase exhibited growth-inhibitory effects that were dependent on the dose against H. pylori (Fig. 1b and c). Androstenedione, however, was less potent than progesterone in inhibiting the growth of H. pylori. The CFUs of H. pylori cultured for 24 h with androstenedione at the 100 μM concentration

were slightly lower than the baseline CFU (105.5 CFU mL−1), whereas the CFUs of the organisms cultured for 24 h with progesterone at the 100 μM concentration were below the limits of detection. Thus, progesterone demonstrated the most effective anti-H. pylori action of the three steroid hormones, and it appeared that this action was bactericidal to H. pylori. This led us to investigate the antibacterial effect of progesterone on H. pylori in more detail. Progesterone has two derivatives: 17α-hydroxyprogesterone (17αPS) and 17α-hydroxyprogesterone caproate (17αPSCE). The derivatives 17αPS and 17αPSCE are modified by a hydroxyl group and an acyl group (caproic acid), respectively, at the carbon 17 position of the progesterone framework (Fig. 2). Noting this, we next examined the anti-H. pylori action of 17αPS and 17αPSCE using the simple-PPLO broth. Surprisingly, 17αPS, a natural progesterone derivative, had no influence on the growth of H. pylori.