Phlebotomy improves histology in chronic hepatitis C males with mild iron overload

Table 1 Epidemiological, clinical and laboratory characteristics at the time of their first liver biopsy of 28 patients with chronic hepatitis C included in the study

Variable1		Range	SD
Age (yr)	58	43-73	7.6
Gender (males/females)	21/7
Body mass index (kg/m2)	24.5	19-38.4	3.9
HCV genotype (1/non-1)	17/11
Drug addiction (yes/no)	2/26
Previous transfusion (yes/no)	11/17
Alcohol abuse (yes/no)2	3/25
Alcohol intake (g/d)	15	0-60	18.35
Treatment before recruitment (interferon based therapy/no treatment)	13/15
Aspartate aminotransferase (U/L, 0-40)	108	46-275	58
Alanine aminotransferase (U/L, 0-40)	130	52-318	57
γ glutamyl transpeptidase (U/L, 0-50)	86	14-242	59
Hemoglobin (g/100 mL, 13.7-17)	14.7	11-18	1.6
Serum iron (μmol/L, 11-32)	24.3	73-240	6.9
Transferrin saturation (%, 20-50)	43	23-69	11
Serum ferritin (ng/mL, 5-365)	387	64-1051	238
Hepatic iron grade (Searle’s score 0/1/2/3)	6/17/15/0	0-2
Hepatic iron concentration (μmol/g dry tissue, < 25)	23.3	7.5-40.5	7
Histological grade (Knodell’s score, median)	5	2-9
Histological stage (Knodell’s score 0/1/3/4)	0/14/9/5	1-4
Advanced fibrosis or cirrhosis (Knodell’s stage score 3-4) (yes/no)	14/14
Steatosis grade (0/1/2/3)3	9/12/5/0	0-2

¹Values are means, counts or medians when indicated;

²Yes: ≥ 50 g/d for men or 20 g/d for women;

³Data were available for 26 out of 28 patients. Steatosis grade: 0, no steatosis; 1, ≤ 33% of hepatocytes; 2, 34%-66%; 3, > 66%.

Table 2 Clinical and laboratory changes between the two liver biopsies of 28 patients with chronic hepatitis C included in the study

Variable	First biopsy1	Second biopsy1	P2
Body mass index (kg/m2)	24.5 ± 3.9	24.5 ± 3.8	0.93
Alcohol intake (g/d)	15 ± 18	7.5 ± 9.7	0.034
HCV-RNA (× 106 copies/mL)3	4.54 ± 3.6	3.96 ± 3.5	0.11
Aspartate aminotransferase (U/L, 0-40)	108 ± 58	64 ± 49	0.0001
Alanine aminotransferase (U/L, 0-40)	130 ± 57	70 ± 43	< 0.0001
γ glutamyl transpeptidase (U/L, 0-50)	86 ± 59	56 ± 56	0.004
Hemoglobin (g/100 mL, 13.7-17)	14.7 ± 1.6	14 ± 1.4	0.0001
Serum iron (μmol/L, 11-32)	24.3 ± 6.9	16.4 ± 6	< 0.0001
Transferrin saturation (%, 20-50)	43 ± 11	24.5 ± 10	< 0.0001
Serum ferritin (ng/mL, 5-365)	387 ± 238	46 ± 51	< 0.0001
Hepatic iron grade (Searle’s score 0/1/2/3)	6/17/5/0	27/1/0/0	< 0.0001
Hepatic iron concentration (μmol/g dry tissue, < 25)	23.3 ± 7	8 ± 5.6	< 0.0001
Histological grade (Knodell’s score, median)	5	4.5	0.43
Histological stage (Knodell’s score 0/1/3/4)	0/14/9/5	0/13/9/6	0.61
Steatosis grade (0/1/2/3)4	9/12/5/0	11/11/3/1	0.41

¹Values are mean ± SD, counts or medians when indicated;

²Paired Student’s t-test or Wilcoxon matched-pairs signed-ranks test;

³Data were available for 20 out of 28 patients;

⁴Data were available for 26 out of 28 patients. Steatosis grade: 0, no steatosis; 1, ≤ 33% of hepatocytes; 2, 34%-66%; 3, > 66%.

Table 3 Individual values of serum aminotransferase, daily alcohol intake, hepatic iron concentration, histological staging and grading scores before (columns a) and at the end of long-term phlebotomy (columns b) in patients with or without evidence of histological improvement

No.	Age (yr)	Gender	Alcohol		BMI		AST		ALT		HIC		Staging		Grading		HI
			a	b	a	b	a	b	a	b	a	b	a	b	a	b
1	53	M	30	20	21.9	21.6	81	57	138	76	7.9	5.1	3	1	4	3	Yes
2	56	M	0	0	20.8	20.4	74	57	114	54	22.4	2.8	3	1	7	2	Yes
3	50	M	40	20	25.5	25.9	183	50	158	36	19.8	4.9	4	4	8	3	Yes
4	59	M	0	0	22.9	23.2	275	194	121	76	24.9	12.1	4	4	5	7	No
5	68	M	0	0	24.0	24.3	104	50	116	78	21.7	5.2	1	1	4	4	No
6	58	F	0	0	21.9	21.6	72	67	117	77	29.3	8.6	3	3	5	5	No
7	67	F	0	0	22.7	22.7	109	88	148	93	17.3	8.8	1	3	3	4	No
8	65	F	0	0	19.6	20.2	155	215	220	195	23.3	7.3	3	4	6	7	No
9	53	M	0	0	22.8	22.4	139	113	159	100	7.5	3.7	4	4	4	6	No
10	51	M	30	30	23.9	23.9	50	32	92	53	40.5	24.8	1	1	4	2	Yes
11	64	F	10	10	38.4	37.7	129	144	147	138	32.5	12.7	4	4	5	7	No
12	52	M	30	10	25.3	24.9	64	54	138	74	30.4	22.8	3	1	4	2	Yes
13	54	M	20	20	24.2	24.2	52	42	94	62	15.8	9.2	1	1	2	3	No
14	65	M	0	0	30.5	30.8	47	34	73	53	22.8	11.8	4	4	6	4	No
15	73	M	10	10	20.9	20.9	140	94	221	120	30.1	3.1	3	1	6	4	Yes
16	42	M	50	30	24.2	23.5	96	22	64	19	28.5	13.1	3	3	9	2	Yes
17	56	M	0	0	23.3	23.6	95	42	103	57	23.3	3.6	3	1	9	9	Yes
18	55	M	0	0	24.5	23.5	103	36	113	40	28.7	3.6	1	1	5	3	Yes
19	43	M	30	10	29.4	28.1	256	61	318	110	21.5	5.4	1	1	9	3	Yes
20	57	M	20	0	27.7	27.7	124	50	135	90	14.3	3.6	1	3	5	7	No
21	50	F	0	0	20.7	21.1	46	37	52	49	25.1	5.4	1	3	5	5	No
22	59	M	60	20	26.9	28.1	96	20	114	22	32.2	1.8	1	1	7	5	Yes
23	62	M	20	0	24.6	24.6	90	30	142	32	19.7	5.4	1	1	3	7	No
24	67	M	50	10	24.8	24.8	52	48	59	54	16.1	5.4	1	3	5	9	No
25	66	M	10	10	23.7	23.3	94	51	128	70	23.3	12.5	3	3	9	7	Yes
26	54	F	0	0	19.3	21.2	64	21	83	23	16.1	3.6	1	1	5	7	No
27	64	F	0	0	29.7	29.7	68	25	85	29	28.7	7.2	1	3	5	7	No
28	51	M	10	10	22.7	22.7	160	50	201	76	17.9	10.7	1	3	4	3	No

Alcohol: Alcohol intake (g/d); BMI: Body mass index; AST: Aspartate aminotransferase (U/L); ALT: Alanine aminotransferase (U/L); HIC: Hepatic iron concentration (μmol/g dry tissue); Staging: Knodell’s score; Grading: Knodell’s score; HI: Histological improvement defined by at least one point reduction in the histological staging score or, in case of unchanged stage, as at least two points reduction in the histological grading score.

Table 4 Univariate and multivariate logistic regression analysis of the putative predictors of the histological improvement in 28 patients with chronic hepatitis C treated with phlebotomy

	Odds ratio (95% CI)	P
Univariate analysis
Age (yr)	0.90 (0.80-1.01)	0.085
Male gender	Females are all non responders
Drug addict (yes/non)1
Previous transfusion (yes/non)	1.19 (0.26-5.5)	0.82
Alcohol abuse (yes/non)2	3 (0.24 38)	0.39
Alcohol intake (g/d)3	1.06 (1.004-1.11)	0.034
Body mass index (kg/m2)	0.96 (0.78-1.17)	0.69
HCV Genotype (1 vs non-1)	0.84 (0.18-3.95)	0.82
Previous antiviral treatment (yes/non)	0.71 (0.16-2.23)	0.66
Days between the two liver biopsies	1 (0.99-1.001)	0.91
Aspartate aminotransferase (U/L)	1.001 (0.99-1.01)	0.79
Alanine aminotransferase (U/L)	1.006 (0.99-1.02)	0.37
γ glutamyl transpeptidase (U/L)	1.002 (0.99-1.01)	0.71
Hemoglobin (g/100 mL)	1.32 (0.79-2.21)	0.28
Serum iron (μmol/L)	0.99 (0.98-1.02)	0.9
Transferrin saturation (%)	0.97 (0.90-1.04)	0.42
Serum ferritin (ng/mL)	1.004 (0.99-1.008)	0.06
Histological grade (Knodell’s score)3	2.4 (1.2-4.95)	0.013
Histological stage (Knodell’s score)	1.32 (0.71-2.45)	0.12
Steatosis grade (0/1/2/3)4	0.7 (0.23-2.1)	0.53
Hepatic iron concentration (μmol/g dry tissue)3	1.16 (1-1.34)	0.043
Hepatic iron grading (Searle’s score)	2.64 (0.69-10.1)	0.15
Multivariate analysis on male patients
Hepatic iron concentration (μmol/g dry tissue)	1.41 (1.03-1.94)	0.031

¹Only two subjects were drug addicts, so the univariate analysis was not performed;

²Yes: ≥ 50 g/d for men or 20 g/d for women;

³These variables were selected to enter in the multivariate model;

⁴Data were available for 26 out 28 patients. Steatosis grade: 0, no steatosis; 1, ≤ 33% of hepatocytes; 2, 34%-66%; 3, > 66%.