Fig. 4.
Brexanolone infusion alters in vitro blood cell responses to the TLR7 inflammatory agonist imiquimod (IMQ) as defined by the levels of cytokines IL-6, IL-1β, and TNF-α. The inhibition of the IMQ-induced IL-6, IL-1β, and TNF-α correlates with HAM-D score improvement after brexanolone infusion. Whole blood drawn from post-partum depression (PPD) patients ∼1 h before and 6 h after 60 h duration of brexanolone (Brex) infusion was exposed to IMQ (30 μg/ml; 37 °C; 4 h) in cell culture in vitro. The levels of IL-6, IL-1β, and TNF-α in culture at baseline, and following the in vitro exposure to IMQ were examined in isolated cell lysates by ELISAs. a: Since the values of the baseline adjusted IMQ-induced levels of the cytokines in the pre- or post-brexanolone condition, did not represent normal (Gaussian) distribution (the Kolmogorov–Smirnov normality test), the Multiple Wilcoxon matched-pairs signed rank tests with Bonferroni-Dunn correction for the multiple comparisons were applied. The median values of differences in picograms/milligram total protein (pg/mg) and % confidence intervals (CI) of differences were examined. The levels of IL-6 (Npairs = 9), IL-1β (Npairs = 9), and TNF-α (Npairs = 9) were significantly reduced in the post-brexanolone (After Brex) condition when compared with the pre-brexanolone (Before Brex) condition. The W values, p and adjusted p values are presented in the figure. ∗p < 0.05, ∗∗p < 0.01, [MLT Wilcoxon BFD]. b: Linear regression analysis was applied to examine whether the % inhibition in the IMQ-induced IL-6, IL-1β, or TNF-α after brexanolone infusion correlated with the % decrease (improvement) in HAM-D score. The coefficient of determination (R2 value), F statistic, degrees of freedom, and p value were examined. The inhibition (%) of the IMQ-induced IL-6, IL-1β, and TNF-α were positively correlated with the % decrease (improvement) in HAM-D score after brexanolone infusion. ∗p < 0.05, [Linear regression analysis].