Fig. 3

High concentrations of 17β-Estradiol (hE2) promote microglial microglial response. A Viability of BV-2 cells after treatment with E2 for various durations and concentrations. B Viability of primary microglia after treatment with E2 for various concentrations. BV2 cells are fixed and immunostained for Iba1 (green) and CD86 (red) (C, F). Bar = 30 µm. Cells are incubated with E2 of 0 nmol/L to 3200 nmol/L for 24 h (C, E) or pre‐treated with 100 ng/mL of LPS for 1 h and then co‐treated with LPS and E2 (200 or 800 nmol/L) for 24 h (F). The mean fluorescence intensity of CD86 or Iba1 and the levels of Iba1 intensity expressed as a relative change in comparison with untreated cells (D, G). E Representative band of protein expression. The mRNA levels of proinflammatory factors IL-1β, IL-6, and TNF-α (H), and anti-inflammatory factors IL-4, IL-10, and TGF-β (i) are evaluated by qPCR. Students t-test is performed to determine the significant difference based on P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***), respectively, in comparison to the control group as indicated by black asterisks or ns and the LPS + E2 (Low) group as indicated by red asterisks. ns: no statistical significance. Data are presented as mean ± the standard deviation (SD). Each experiment is repeated independently in three