Pro12Ala polymorphism of the peroxisome proliferator-activated receptor γ2 in patients with fatty liver diseases

Table 1 Scoring according to the histological features described by Kleiner et al[15]

Definition	Score
Steatosis
Grade
< 5%	0
5%-33%	1
33%-66%	2
> 66 %	3
Localization
Zone 3	0
Zone 2	1
Zone 1	2
Azonal	3
Panacinar	4
Type
Macrovesicles	In %
Microvesicles	In %
Mixed
Inflammation
Lobular
No foci	0
< 2 foci	1
2-4 foci	2
> 4 foci	3
Portal
No inflammation	0
Minimal	1
Mild	2
Moderate	3
Severe	4
Fibrosis
None	0
Mild/moderate	1
Periportal or perisinusoidal	2
Bridging fibrosis	3
Cirrhosis	4
Liver cell damage
Ballooning
None	0
Moderate	1
Severe	2
Mallory bodies
None	0
Moderate	1
Severe	2

Table 2 Real-time polymerase chain reaction primers and probes

Name	Oligonucleotide sequence	PCR application
Plas A primer	5'-CCGCTCTAGAACTAGTGAAGGAA-3'	Reference DNA
Plas S primer	5'-ACTCACTATAGGGCGAATTGG-3'
PPARγ A primer	5'-TTACCTTGTGATATGTTTGCAGACA-3'	Target DNA
PPARγ mis primer	5'-GTTATGGGTGAAACTCTTGGAGA-3'
TM LNA probe wt	5'-6FAM-CTATTGACCCAGAAAGC--BHQ1	Target and reference DNA
TM LNA probe mut	5'-YAK-CTATTGACGCAGAAAGC--BHQ1

Bold letters indicate the single nuclear polymorphism and underlined letters indicate the insertion of locked nucleotides (LNA). PPARγ: Peroxisome proliferator-activated receptor-γ; PCR: Polymerase chain reaction.

Table 3 Occurrence of diabetes mellitus in patients with fatty liver disease n (%)

	No C2 (NAFLD)	C2 (AFLD)
Diabetes mellitus positive	33 (12.5)	14 (14)
Diabetes mellitus negative	144 (54.8)	51 (51)
ND	86 (32.7)	35 (35)
Total	263 (100)	100 (100)

NAFLD: Non-alcoholic fatty liver disease; AFLD: Alcoholic fatty liver disease; ND: Not determined.

Table 4 Histological features in the fatty liver diseases cohort

Cases of fatty liver disease (n = 363)	n (%)
Steatosis
S0	9 (2.5)
S1	169 (46.6)
S2	84 (23.1)
S3	101 (27.8)
Ballooning
No ballooning	164 (45.2)
Stage 1	124 (34.2)
Stage 2	75 (20.7)
Inflammation
G0	4 (1.1)
G1	217 (59.8)
G2	113 (31.1)
G3	29 (8)
Fibrosis
F0	97 (26.7)
F1	116 (32)
F2	66 (18.2)
F3	56 (15.4)
F4	28 (7.7)

Table 5 Occurrence of the wt and the mutated form of peroxisome proliferator-activated receptor-γ2 depending on the grade of steatosis and ballooning n (%)

	Steatosis						Ballooning
	NAFLD			AFLD			NAFLD			AFLD
	Total	Moderate (0-1)	Severe (2-3)	Total	Moderate (0-1)	Severe (2-3)	Total	-	+	Total	-	+
Allelic discrimination
Mutation	66 (100)	27 (41)	39 (59)	26 (100)	12 (46)	14 (54)	66 (100)	28 (42)	38 (58)	26 (100)	19 (73)	7 (27)
Wild type	460 (100)	229 (50)	231 (50)	174 (100)	88 (51)	86 (49)	460 (100)	206 (45)	254 (55)	174 (100)	121 (70)	53 (30)
P value	0.112			0.417			0.411			0.454
Odds ratio (CI)	1.43 (0.85-2.42)			1.19 (0.52-2.73)			1.10 (0.65-1.85)			0.84 (0.33-2.12)

NAFLD: Non-alcoholic fatty liver disease; AFLD: Alcoholic fatty liver disease.

Table 6 Occurrence of the wt and mutated form of the peroxisome proliferator-activated receptor-γ2 depending on the grade of inflammation and fibrosis n (%)

	Inflammation						Fibrosis
	NAFLD			AFLD			NAFLD			AFLD
	Total	Moderate (0-1)	Severe (2-3)	Total	Moderate (0-1)	Severe (2-3)	Total	F0-F1	F2-F4	Total	F0-F1	F2-F4
Allelic discrimination
Mutation	66 (100)	38 (58)	28 (42)	26 (100)	9 (35)	17 (65)	66 (100)	47 (71)	19 (29)	26 (100)	4 (15)	22 (85)
Wild type	460 (100)	296 (64)	164 (36)	174 (100)	99 (57)	75 (43)	460 (100)	327 (71)	133 (29)	174 (100)	54 (31)	120 (69)
P value	0.175			0.028			0.555			0.075
Odds ratio (CI)	1.33 (0.79-2.25)			2.50 (1.05-5.90)			0.99 (0.56-1.76)			2.48 (0.81-7.53)

NAFLD: Non-alcoholic fatty liver disease; AFLD: Alcoholic fatty liver disease.