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

Table 1 Characteristics at the time of isolation of PBMCs from age-matched younger and older males and pre- and post-menopausal females of patients with chronic hepatitis C and healthy controls

	Patients with chronic hepatitis C				Healthy controls
	Males (yr)		Females (yr)		Males (yr)		Females (yr)
	< 50	≥ 50	< 50	≥ 50	< 50	≥ 50	< 50	≥ 50
n	18	18	17	18	18	18	18	18
Age (yr)	41 ± 7	69 ± 6	41 ± 7	68 ± 6	40 ± 6	68 ± 6	40 ± 6	69 ± 6
HCV-RNA (log copies/mL)	6.1 ± 0.8	6.5 ± 1.4	5.9 ± 0.8	6.4 ± 1.2	ND	ND	ND	ND
ALT (U/L)	118 ± 68	112 ± 52	105 ± 49	101 ± 55	27 ± 8	26 ± 8	24 ± 7	24 ± 8
Platelet count (103/mm3)	208 ± 58	189 ± 52	206 ± 54	185 ± 55	230 ± 60	219 ± 53	225 ± 57	214 ± 58

The PBMCs were isolated from age-matched males (n = 72) and females (n = 71) of 71 patients with chronic hepatitis C and 72 healthy controls, who were divided into younger or pre-menopausal (< 50 years of age) and older or post-menopausal (≥ 50 years of age) groups, based on a mean menopausal age of 50 years. The values are the mean ± SD (n = 17 or 18). ND: Not detected.

Table 2 Spontaneous production of TNF-α, IL-1β, IL-8, and MCP-1 by PBMCs from age-matched younger and older males and pre- and post-menopausal females of patients with chronic hepatitis C and healthy controls (pg/mL supernatant)

Subjects with and without chronic HCV infection			TNF-α	IL-1β	IL-8	MCP-1
Males	Younger	Patients	354 ± 201a	253 ± 142	918 ± 499	403 ± 216
	(< 50 yr)	Controls	265 ± 161	180 ± 105	780 ± 420	308 ± 161
	Older	Patients	385 ± 234a	284 ± 160a	1087 ± 576a	461 ± 242a
	(≥ 50 yr)	Controls	322 ± 200	217 ± 125	899 ± 480	364 ± 187
Females	Pre-menopausal	Patients	288 ± 177	198 ± 111	780 ± 410	334 ± 174
	(< 50 yr)	Controls	210 ± 126	167 ± 94	624 ± 335	275 ± 140
	Post-menopausal	Patients	314 ± 189	264 ± 149a	887 ± 473	387 ± 198
	(≥ 50 yr)	Controls	241 ± 148	182 ± 100	699 ± 372	362 ± 201

The PBMCs were isolated from age-matched males (n = 72) and pre- and post-menopausal females (n = 71) of 71 patients with chronic hepatitis C and 72 healthy controls. The levels of TNF-α, IL-1β, IL-8, and MCP-1 in the culture supernatant were detected by means of an ELISA. The values are the mean ± SD (n = 17 or 18).

^aP < 0.05 vs the pre-menopausal healthy controls.

Table 3 Effects of E2 and progesterone on spontaneous production of TNF-α, IL-1β, IL-8, and MCP-1 by unstimulated PBMCs from age-matched older male and post-menopausal female patients with chronic hepatitis C (% of initial value)

Chronic hepatitis C subjects	TNF-α	IL-1β	IL-8	MCP-1
Older male patients
None	123 ± 14	120 ± 16	119 ± 12	114 ± 15
+ E2	82 ± 10a	84 ± 10a	80 ± 11a	78 ± 11a
+ E2 + ICI	118 ± 16	114 ± 18	108 ± 19	99 ± 19
+ Progesterone	167 ± 19a	169 ± 20a	159 ± 18a	169 ± 28a
+ Progesterone + RU	129 ± 21	124 ± 21	132 ± 19	122 ± 24
Post-menopausal female patients
None	118 ± 15	119 ± 14	122 ± 16	110 ± 13
+ E2	83 ± 11a	79 ± 9a	81 ± 12a	74 ± 11a
+ E2 + ICI	108 ± 14	122 ± 16	117 ± 19	104 ± 19
+ Progesterone	167 ± 13a	170 ± 24a	159 ± 20a	175 ± 30a
+ Progesterone + RU	130 ± 27	125 ± 28	132 ± 30	123 ± 25

The unstimulated PBMCs from the age-matched older male and post-menopausal female patients with chronic hepatitis C were cultured for another 6 h with and without 10^-8 mol/L E2 (E2) or 10^-7 mol/L progesterone (progesterone) in the presence and absence of 10^-6 mol/L ICI (ICI) or 10^-6 mol/L RU (RU). The levels of TNF-α, IL-1β, IL-8, and MCP-1 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 and receptor antagonists. The values are the mean ± SD (n = 18).

^aP < 0.05 vs 6-h-cultures in the absence of the female sex hormones and receptor antagonists.

Table 4 Effects of E2 and progesterone on hydrogen peroxide-stimulated production of TNF-α, IL-1β, IL-8, and MCP-1 by PBMCs from age-matched younger male and pre-menopausal female healthy controls (pg/mL supernatant)

Healthy subjects	TNF-α	IL-1β	IL-8	MCP-1
Younger male controls
Oxidative stress	102 ± 17	66 ± 11	218 ± 41	474 ± 81
Oxidative stress + E2	78 ± 11a	49 ± 6a	174 ± 27a	332 ± 49a
Oxidative stress + E2 + ICI	107 ± 10	62 ± 11	223 ± 46	457 ± 87
Oxidative stress + Progesterone	140 ± 14a	86 ± 9a	289 ± 40a	658 ± 98a
Oxidative stress + Progesterone + RU	110 ± 23	75 ± 14	223 ± 32	516 ± 73
Pre-menopausal female controls
Oxidative stress	91 ± 15	57 ± 9	195 ± 37	435 ± 85
Oxidative stress + E2	64 ± 11a	42 ± 6a	155 ± 29a	314 ± 68a
Oxidative stress + E2 + ICI	95 ± 13	54 ± 9	187 ± 37	419 ± 92
Oxidative stress + Progesterone	129 ± 12a	75 ± 12a	255 ± 40a	586 ± 70a
Oxidative stress + Progesterone + RU	101 ± 19	63 ± 11	208 ± 40	462 ± 73

The unstimulated PBMCs from the age-matched younger male (n = 18) and pre-menopausal female (n = 18) healthy controls were incubated for up to 6 h in serum-free RPMI after exposure to 10^-5 mol/L hydrogen peroxide (oxidative stress) with and without 10^-8 mol/L E2 (E2) or 10^-7 mol/L progesterone (Progesterone) in the presence and absence of 10^-6 mol/L ICI (ICI) or 10^-6 mol/L RU (RU). The levels of TNF-α, IL-1β, IL-8, and MCP-1 in the culture supernatant incubated for 6 h were detected by means of an ELISA. The values are the mean ± SD (n = 18).

^aP < 0.05 vs 1 h cultures for MCP-1 or 6 h cultures for TNF-α, IL-1β, and IL-8 after hydrogen peroxide exposure in the absence of the female sex hormones and receptor antagonists.