Effects of estradiol and progesterone on the proinflammatory cytokine production by mononuclear cells from patients with chronic hepatitis C

Abstract

Aim: To investigate the effects of estradiol (E2) and progesterone on the unstimulated and oxidative stress-stimulated production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-8, and macrophage chemotactic protein (MCP)-1 by peripheral blood mononuclear cells (PBMCs) from patients with chronic hepatitis C and healthy controls.

Methods: The PBMCs were separated from age-matched 72 males and 71 females with and without chronic hepatitis C, who were divided into two groups based on a mean menopausal age of 50 years. Oxidative stress was induced by hydrogen peroxide in the cells incubated in serum-free media. Cytokines in the culture supernatant were measured by an enzyme-linked immunosorbent assay.

Results: The highest levels of the spontaneous production of TNF-alpha, IL-1beta, IL-8, and MCP-1 by the unstimulated PBMCs were in the older male patients with chronic hepatitis C and the lowest levels were in the pre-menopausal female healthy controls. E2 inhibited the cytokine production by the unstimulated PBMCs from the older male and post-menopausal female patients, which was further stimulated by progesterone. The exposure to hydrogen peroxide in the PBMCs from the younger male and pre-menopausal female healthy subjects induced the production of cytokines. The change rates of the hydrogen peroxide-stimulated cytokine production were suppressed by E2 and enhanced by progesterone.

Conclusion: These findings suggest that E2 may play a favorable role in the course of persistent liver injury by preventing the accumulation of monocytes-macrophages and by inhibiting proinflammatory cytokine production, whereas progesterone may counteract the favorable E2 effects.

Figure 1

Effects of E2 and progesterone on spontaneous production of TNF-α (A), IL-1β (B), IL-8 (C), and MCP-1 (D) by unstimulated PBMCs from age-matched older male and post-menopausal female patients with chronic hepatitis C. The unstimulated PBMCs from the age-matched older male (black square) and post-menopausal female (black circle) patients with chronic hepatitis C were cultured for another 6 h with and without E2 (10^-10-10^-7 mol/L) (solid) or progesterone (10^-10-10^-7 mol/L) (open). The spontaneous production levels of TNF-α (A), IL-1β (B), IL-8 (C), and MCP-1 (D) in the culture supernatant were detected by means of an ELISA. The results were expressed as the percentages of each initial value for the cytokine production in the absence of the female sex hormones. The values are the mean ± SD (n = 18). ^aP < 0.05 in comparison to the 6-h-cultures in the absence of the female sex hormones.

Figure 2

Stimulation of TNF-α, IL-1β (A), IL-8, and MCP-1 (B) production after exposure to hydrogen peroxide by unstimulated PBMCs from age-matched younger male and pre-menopausal female healthy controls. The unstimulated PBMCs from the age-matched younger male (black square) and pre-menopausal female (black circle) healthy controls were incubated for up to 6 h in serum-free RPMI in the presence of 10^-5 mol/L hydrogen peroxide. The levels of TNF-α (A solid), IL-1β (A open), IL-8 (B solid), and MCP-1 (B open) in the culture supernatant were detected by means of an ELISA. The values are the mean ± SD (n = 10).