Serum Bcl-2 concentrations in overweight-obese subjects with nonalcoholic fatty liver disease

Table 1 Clinical characteristics and laboratory data of the study population (mean ± SD)

	Diagnosis
	H	FL	NASH	P value
Subjects (n)	21	43	41
Females	11	30	24	0.951
Age (yr)	19.1 ± 3.8	31.4 ± 13.9	35.6 ± 13	< 0.0012
Overweight	0	3	3	0.71
Obesity 1st grade	0	13	9	0.51
Obesity 2nd grade	0	9	6	0.61
Obesity 3rd grade	0	18	24	0.851
MS	0	24	36	0.0015
MUO	0	34	39	0.061
BMI
F	23.8 ± 1.13	38.8 ± 8.24	37.8 ± 6.6	< 0.0012
M	23.7 ± 0.6	38.6 ± 4.4	45.7 ± 10.5	0.013
WC (cm)
F	86.3 ± 1.4	114.8 ± 15.7	117.3 ± 17.7	0.0012
M	93.4 ± 2.3	121.2 ± 11.3	131.1 ± 15.1	< 0.0013
HOMA [median (range)]	1.77 (1.03-6)	2.3 (1.4-7.3)	4.1 (0.6-10.2)	< 0.00013
γ-GT U/L [median (range)]	33.1 (23.4-44.8)	40.5 (26-80)	54 (22.5-397)	< 0.00019
ALT U/L [median (range)]	27 (18-34)	36 (14-133)	40 (11-153)	0.0046
Uric acid	3.5 ± 0.4	5.1 ± 1.45	5.4 ± 1.4	< 0.0012
Ferritin
F	84.6 ± 10.3	139.6 ± 76.	149.2 ± 69	0.0012
M	166.8 ± 37.4	218.5 ± 121.	280 ± 103	0.014
Triglycerides	85 (66-121)	96 (51-290)	117 (51-386)	0.1610
HDL (mg/mL)
F	63.6 ± 7.97	51.8 ± 9.38	50 ± 11.15	0.0012
M	55.3 ± 8.4	50.8 ± 7.9	44 ± 4.4	< 0.0018
Triglycerides/HDL [median (range)]
F	1.32 (1.16-1.49)	2.19 (1.18-6.5)	1.51 (1.07-7.6)	0.0046
M	1.635 (1.39-1.75)	1.67 (1.25-2.7)	2.65 (1.25-8.8)	0.037

¹χ² test between fatty liver (FL) and non alcoholic steato hepatitis (NASH);

²analysis of variance (ANOVA) [significance between healthy (H) and FL and between H and NASH];

³ANOVA (significance between H and FL, between H and NASH and also between FL and NASH);

⁴ANOVA (significance between H and NASH);

⁵Pearson χ² (significance between FL and NASH);

⁶Kruskal-Wallis test (significance between H and FL and between H and NASH);

⁷Kruskal-Wallis test (significance between NASH and FL and between NASH and H);

⁸ANOVA (significance between NASH and FL and between NASH and H);

⁹Kruskal-Wallis test (significance between H and FL, between H and NASH and also between FL and NASH);

¹⁰Kruskal-Wallis test. F: Females; M: Males; n: Number; H: Healthy; FL: Fatty liver; NASH: Non alcoholic steato hepatitis; ALT: Alaninamino transferase; γ-GT: γ-glutamil transferase; HDL: High density lipoprotein; HOMA: Homeostatic metabolic assessment; BMI: Body mass index; MS: Metabolic syndrome; MUO: Metabolically unhealthy overweight/obese; WC: Waist circumference.

Table 2 Serum concentrations of the anti-apoptotic protein Bcl-2, histology features and inflammation parameters of the patients enrolled

	Diagnosis
	H	FL	NASH	P value
Subjects (n)	21	43	41
Bcl-2 (ng/mL)	2.5 (1.4-13)	25 (2.5-168)	14 (2.5-89)	< 0.0013
Hepatocyte ballooning grade	NA	NA	2 (1-3)	NA
Steatosis score	NA	2 (1-3)	2 (1-3)	0.492
Lobular inflammation grade	NA	NA	2 (1-3)	NA
Portal inflammation grade	NA	NA	1 (1-3)	NA
Fibrosis score	NA	NA	1 (1-2)	NA
Apoptotic cells (n/mm2)	NA	290 (120-900)	780 (110-1650)	< 0.00012
HS at US grade	NA	3 (0-3)	3 (0-3)	0.0642
CRP (mg/L)	0.9 (0.4-5.4)	3.9 (0.1-21.1)	5 (0.1-27)	0.00011

¹Kruskal-Wallis test [significance between healthy (H) and fatty liver (FL) and between H and non alcoholic steato hepatitis (NASH)];

²Mann-Whitney test;

³Kruskal-Wallis test (significance between H and FL, between H and NASH and also between FL and NASH). n: Number; H: Healthy; FL: Fatty liver; NASH: Non alcoholic steato hepatitis; NA: Not applicable; HS: Hepatic steatosis at ultrasound; US: Ultrasound; SLD: Spleen longitudinal diameter at ultrasound; CRP: C-reactive protein.

Table 3 Prediction of anthropometric parameters, apoptosis detection and laboratory data by Bcl-2 concentrations

Effect	β	t	P value	Tolerance
HOMA	-0.505	-4.27	0.000	0.440
HDL	0.22	1.4	0.16	0.242
WC	0.25	1.7	0.09	0.288
BMI	0.35	2.65	0.009	0.344
Triglycerides	-0.07	-0.17	0.86	0.040
Triglycerides/HDL	0.22	0.5	0.63	0.029
Uric acid	0.24	2.1	0.03	0.509
Ferritin	-0.15	-1.8	0.08	0.853
Apoptosis in NASH	0.013	0.13	0.9	1

NASH: Nonalcoholic steatohepatitis; BMI: Body mass index; HOMA: Homeostatic metabolic assessment; HDL: High density lipoprotein; triglycerides and triglycerides/HDL were considered variables affected by collinearity; WC: Waist circumference; β: Beta, standardized coefficient; t: A measure of the precision with which the regression coefficient is measured.

Table 4 Hidden relationships highlighted by factor analysis

Factors	1	2
Bcl-2	-0.105	0.672
Age	-0.348	-0.424
Apoptosis	0.293	-0.425
MS	0.606	0.011
BMI	0.564	0.528
WC	0.604	0.467
HOMA	0.781	-0.211
HDL	-0.715	0.107
Triglycerides	0.521	-0.471
Triglycerides /HDL	0.698	-0.468
Uric Acid	0.554	0.520
Ferritin	0.279	0.064
ALT	0.545	0.097
Γ-GT	0.074	-0.384
CRP	0.717	-0.274
SLD	-0.202	0.282
HS at US	0.471	0.255
Steatosis	0.293	0.265

Insulin resistance is closely related to inflammation and lipid asset; apoptosis does not seem related to age. Percent of total variance explained for factor 1:26; for factor 2:14; the critical value is 0.568. HS: Hepatic steatosis; US: Ultrasound; SLD: Spleen longitudinal diameter at ultrasound; CRP: C-reactive protein; BMI: Body mass index; MS: Metabolic syndrome; ALT: Alaninamino transferase; γ-GT: γ-glutamyl transferase; HDL: High density lipoprotein; HOMA: Homeostatic metabolic assessment; WC: Waist circumference.