Dual effect of the GHRL gene variant in the molecular pathogenesis of obesity

Details of polymerase chain reaction primers and restriction endonucleases used, and of the resulting restriction fragments_

GHRL Polymorphism	PCR Primers (5’>3’)	Restriction Enzyme	Digestion Pattern	Ref.
rs34911341 (Arg51Gln)	F: TCC AGC CTG CCA CTT AGCR: GGA CCC TGT TC ACT GCC AC	SacI	C allele: 373 bp; T allele: 210 and 163 bp	[28]
rs696217 (Leu72Met)	F: TCC AGC CTG CCA CTT AGCR: GGA CCC TGT TC ACT GCC AC	BsrI	G allele: 373 bp; T allele: 271 and 101 bp	[28]

Baseline characteristics of the studied population_

Parameters	Obese Subjects (n=106)	Non Obese Subjects (n=95)	p Value
Age (years)	41.56±9.87	39.03±9.45	0.066
BMI (kg/m2)	41.58±4.93	22.61±1.82	<0.0001
Waist circumference (cm)	112.40±13.13	85.50±7.33	<0.0001
Hip circumference (cm)	120.50±12.57	99.10±6.14	<0.0001
Fasting glucose (mg/dL)	102.5±23.46	89.60±7.35	<0.0001
Total cholesterol (mg/dL)	231.30±38.17	203.90±35.57	<0.0001
LDL-C (mg/dL)	144.30±33.42	125.70±30.29	<0.0001
HDL-C (mg/dL)	47.40±10.55	54.80±9.36	<0.0001
Triglycerides (mg/dL)	163.10±85.11	102.20±41.22	<0.0001
HOMA-IR	4.60±4.01	2.20±1.13	<0.0001

Anthropometric and metabolic characteristics of all subjects in the genotypes of the rs696217 (G>T) polymorphism_ [Data are expressed as mean±SD_ Analysis of variance (ANOVA) was performed for comparison of the subgroups; bold p values are significant_]

Parameters	GG (n=94)	GT (n=91)	TT (n=16)	p Value
Age (years)	40.17±8.48	40.30±11.13	41.88±8.43	0.8
BMI (kg/m2)	34.00±24.48	31.30±8.68	33.13±9.44	0.81
Waist circumference (cm)	95.61±15.70a	102.90±17.43	105.50±20.83	0.005
Hip circumference (cm)	106.70±13.87b,c	113.10±14.08	116.60±17.23	0.002
Fasting glucose (mg/dL)	93.83±12.75	97.62±17.88	105.20±41.12	0.06
Total cholesterol (mg/dL)	221.00±34.66	212.90±43.36	233.40±44.09	0.104
LDL-C (mg/dL)	134.5.0±29.26	134.30±35.43	148.50±41.23	0.266
HDL-C (mg/dL)	52.94±10.61	48.62±10.56d	52.25±9.16	0.018
Triglycerides (mg/dL)	129.00±64.40	137.80±80.77	145.80±92.20	0.59
HOMA-IR	3502.00±2.20	3246.00±2.40	4522.00±3.90	0.34

The genotype distribution and the allele frequencies of the GHRL gene variants_ [Results are presented as n (%)]

A) Genotype	Obese Subjects (n=106)	Non Obese Subjects (n=95)	p Value
rs34911341 (C>T) (Arg51Gln)
CC	104 (98.12)	95 (100.00)	0.40
CT	1 (0.94)	0 (0.00)
TT	1 (0.94)	0 (0.00)
rs696217 (G>T) (Leu72Met)
GG	37 (34.91)	57 (60.00)	0.0012
GT	57 (53.77)	34 (35.79)
TT	12 (11.32)	4 (4.21)
B) Allele Frequency	Obese Subjects (n=106)	Non Obese Subjects (n=95)	p Value
rs34911341 (C>T) (Arg51Gln)
C	209 (98.58)	190 (100.00)	0.25
T	3 (1.42)	0 (0.00)
rs696217 (G>T) (Leu72Met)
G	131 (61.79)	148 (77.79)	0.0005
T	81 (38.21)	42 (22.11)