Tumor necrosis factor-α -G308A polymorphism is associated with liver pathological changes in hepatitis C virus patients

Table 1 Clinical data of 120 chronic hepatitis C virus (F0-F4) patients

	HCV patients with different fibrosis grade (n = 120)
	F0 (n = 30)	F1 (n = 30)	F2-3 (n = 30)	F4 (n = 30)
Age (yr)	38.4 ± 7.1	41.5 ± 8.7	43.5 ± 8.4	48.3 ± 9.3
Sex (female/male)	13/17	12/18	10/20	9/21
BMI (kg/m2)	25.5 ± 2.3	27.6 ± 3.4	28.7 ± 2.7	29.5 ± 2.7
ALT (U/L)	23.4 ±7.1	39.8 ± 23.7	51.1 ± 20.7	60.1 ± 20.3
AST (U/L)	22.4 ± 6.5	34.9 ± 20.9	48.9 ± 25.9	58.7 ± 18.6
ALP (U/L)	113.6 ± 32.8	122.5 ± 37.1	158.7 ± 47.7	153.8 ± 53.5
Total bilirubin (mg/dL)	0.53 ± 0.32	0.61 ± 0.23	0.96 ± 0.37	1.60 ± 0.65
Albumin (g/dL)	4.10 ± 0.33	4.00 ± 0.38	3.80 ± 0.45	3.50 ± 0.39
Platelets (× 109/L)	300.5 ± 56.2	250.2 ± 48.4	239.6 ± 55.6	159.4 ± 69.5
HCV RNA (× 103 IU/mL)	684.3 ± 124.8	860.1 ± 1042	1241.5 ± 1286.7	1501.4 ± 1661.0
Fibroscan	4.70 ± 0.75	6.30 ± 0.36	9.90 ± 1.78	23.27 ± 9.30
Activity
A0-A1	20 (66.7)	13 (43.3)	10 (33.3)	1 (3.3)
A2-A3	10 (33.3)	17 (56.7)	20 (66.7)	29 (96.7)
Steatosis
≤ 33%	28 (93.3)	26 (86.7)	16 (53.3)	4 (13.3)
> 33%	2 (6.7)	4 (13.3)	14 (46.7)	26 (86.7)

The clinical data was compared in HCV patients with different fibrosis grades: F0 (n = 30), F1 (n = 30), F2-F3 (n = 30), F4 (n = 30). The quantitative data were represented as mean ± SD while qualitative data were represented as n (%). HCV: Hepatitis C virus; BMI: Body mass index; ALT: Alanine aminotransferase; AST: Aspartate transaminase; ALP: Alkaline phosphatase.

Table 2 Distribution of tumor necrosis factor α -G308A polymorphism in controls and hepatitis C virus infected patients with different fibrosis grades (F0-F4) n (%)

Polymorphism of TNFα -308	Controls (n = 60)	HCV Patients (n = 120)	F0 patients (n = 30)	F1 patients (n = 30)	F2-3 patients (n = 30)	F4 patients (n = 30)
GG genotype	40 (66.6)	48 (40.0)	18 (60.0)	13 (43.3)	10 (33.3)	7 (23.3)
GA genotype	18 (30.0)	57 (47.5)	10 (33.3)	15 (50.0)	16 (53.4)	16 (53.4)
AA genotype	2 (3.3)	15 (12.5)	2 (6.7)	2 (6.7)	4 (13.3)	7 (23.3)
GA + AA	20 (33.3)	72 (60.0)	12 (40.0)	17 (56.7)	20 (66.6)	23 (76.6)
G allele	98 (81.7)	153 (63.8)	46 (76.7)	41 (68.3)	36 (60.0)	30 (50.0)
A allele	22 (18.3)	87 (36.3)	23 (23.3)	19 (31.7)	24 (40.0)	30 (50.0)

Genotyping of TNFα -G308A was conducted and distribution of different genotypes and alleles was calculated as percentage and compared in 60 controls and 120 chronic HCV patients with different fibrosis grades: F0 (n = 30), F1 (n = 30), F2-F3 (n = 30), F4 (n = 30). TNFα: Tumor necrosis factor alpha; HCV: Hepatitis C virus.

Table 3 Tumor necrosis factor α serum level in 120 hepatitis C virus patients

	TNFαserum levels				P value
	GG	GA	AA	GA + AA	GG vs AA
All fibrosis patients (F0-F4, n = 120)	9.1 ± 2.6	12.1 ± 4.9	16.1 ± 4.9	13.1 ± 5.1	0.0001
early fibrosis patients (F0-F1, n = 60)	8.3 ± 2.6	10.1 ± 4.5	16.2 ± 4.02	10.9 ± 4.9	0.010
late fibrosis patients (F2-F4, n = 60)	10.4 ± 2.1	13.8 ± 4.5	16.1 ± 5.6	14.4 ± 4.9	0.002

The TNFα serum level produced by different TNFα -308 genotypes GG, GA and AA were detected in early and late HCV fibrosis patients. The TNFα serum level is represented as mean ± SD and was statistically compared in patients with genotype GG to those with AA genotype. P≤ 0.05 is considered significant, while P≤ 0.01 is highly significant. TNFα: Tumor necrosis factor alpha; HCV: Hepatitis C virus.

Table 4 Influence of tumor necrosis factor α -308 genotypes on the liver biochemical and pathological parameters

	TNF genotypes (n = 120)			P value
	GG (n = 48)	AG (n = 57)	AA (n = 15)	GG vs AA	GG vs GA + AA
AST (U/L)	31.9 ± 20.6	44.7 ± 21.6	57.7 ± 28.7	0.0003	0.0003
ALT (U/L)	34.5 ± 20.9	46.5 ± 20.5	62.0 ± 28.2	0.0001	0.0004
ALP (U/L)	113.7 ± 36.8	138.3 ± 41.5	190.1 ± 46.5	0.0020	0.3500
Albumin (g/dL)	3.9 ± 0.46	3.9 ± 0.39	3.6 ± 0.59	0.0100	0.2000
Total bilirubin (mg/dL)	0.73 ± 0.41	0.89 ± 0.51	1.3 ± 0.68	0.0002	0.0100
Platelets (× 109/L)	254.9 ± 66.2	224.5 ± 72.3	205.8 ± 96.6	0.0360	0.0220
HCV RNA (× 103 IU/mL)	862.6 ± 1295.6	1083.9 ± 1187.2	1675.5 ± 1477.6	0.0450	0.2200
Activity
A0-A1 (n = 44)	25 (56.8)	18 (40.9)	1 (2.3)	0.0290	0.0350
A2-A3 (n = 76)	23 (30.3)	39 (51.3)	14 (18.4)
Steatosis
≤ 33% (n = 74)	36 (48.7)	34 (45.9)	4 (5.4)	0.0280	0.0300
> 33% (n = 46)	12 (26.1)	23 (50.0)	11 (23.9)
Fibrosis
F0-F1 (n = 60)	28 (46.7)	30 (50.0)	2 (3.3)	0.0310	0.0400
F2-F4 (n = 60)	20 (33.3)	27 (45.0)	13 (21.6)

The effect of different TNFα -308 genotypes (GG, GA, AA) on biochemical parameters (ALT, AST, ALP, albumin, total bilirubin), platelets count and pathological parameters (activity, steatosis, fibrosis) was statistically analyzed. The biochemical data were represented as mean ± SD, while pathological data were represented as n (%). P≤ 0.05 is considered significant, while P≤ 0.01 is highly significant. TNFα: Tumor necrosis factor alpha; HCV: Hepatitis C virus; ALT: Alanine aminotransferase; AST: Aspartate transaminase; ALP: Alkaline phosphatase.

Table 5 Stepwise logistic regression analysis for the effect of tumor necrosis factor α -308 genotypes and alleles on liver disease progression

TNF genotype	Regression coefficient	SE	OR (95%CI)	P value
Activity
GG vs GA + AA	1.329	0.507	3.776 (1.399-10.194)	0.009
G vs A allele	1.171	0.400	3.226 (1.474-7.060)	0.003
Steatosis
GG vs GA + AA	1.502	0.580	4.491 (1.441-14.000)	0.010
G vs A allele	1.099	0.373	3.000 (1.445-6.228)	0.003
Fibrosis
GG vs GA + AA	1.044	0.493	2.841 (1.080-7.472)	0.034
G vs A allele	0.895	0.366	2.446 (1.195-5.008)	0.014

P≤ 0.05 is considered significant while P≤ 0.01 is highly significant. TNF: Tumor necrosis factor.