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. 2019 Dec 5;10:2815. doi: 10.3389/fimmu.2019.02815

Figure 3.

Figure 3

In vitro 17β-estradiol induces cytokines secretion in HiBECs. HiBECs were stimulated with 3 different concentrations of 17β-estradiol (0.1, 10, and 100 nM) for four time-points (0, 12, 24, and 48 h). Total RNA was isolated and analyzed by real-time PCR for IL-6 mRNA, IL-8mRNA and TNF-α mRNA expression. (A–C) The resulted shown that the levels of IL-6mRNA (A), IL-8mRNA (B), and TNF-α mRNA (C) were not changed, when compared with controls at 0, 12, and 24 h. (D–F) The expression of IL-6mRNA (D), IL-8mRNA (E), and TNF-α mRNA (F) were up-regulated at 48 h. And, 10 nM 17β-estradiol can up-regulate expression of IL-6mRNA, IL-8 mRNA, and TNF-α mRNA. *p < 0.05, **p < 0.01, compared with controls by Student t-test. (G–I) Elisa analysis of IL-6, IL-8 and TNF-α protein in cellular supernatants from 17β-estradiol-induced HiBECs at 48 h. **p < 0.01, ***p < 0.001, compared with controls by Student t-test. (J) Immunoblotting analysis of ERα and ERβ expression in HiBECs that were stimulated with 17β-estradiol (10 nM) at four time points (0, 12, 24, and 48 h). (K) 17β-estradiol (10 nM) had no effect on ERα expression at 0, 12, and 24 h, but up-regulated ERα expression at 48 h, **p < 0.01 compared with controls by Student t-test. (L) 17β-estradiol (10 nM) had no effect on ERβ expression at four time points. All data are representative of three independent experiments.