Relationship between glucagon and metabolic dysfunction-associated steatotic liver disease in patients with type 2 diabetes mellitus

Table 1 Anthropometric parameters and biochemical indexes among subjects with or without metabolic dysfunction-associated steatotic liver disease

Variables	UAP ≤ 244 dB/m (n = 68)	UAP > 244 dB/m (n = 144)	P value
Male/female	42/26	96/48	0.538
Age (years)	59 (54, 69)	54 (39, 66)	0.003
Diabetes duration (years)	8.5 (3, 12.75)	5 (1, 10)	0.1
BMI (kg/m2)	22.14 (20.22, 24.21)	26.1 (23.67, 28.65)	< 0.001
WC (cm)	84.32 ± 9.9	92.49 ± 11.24	< 0.001
WHR	0.9 (0.86, 0.95)	0.95 (0.9, 1.0)	< 0.001
SBP (kPa)	18.53 (16.13, 20.13)	18 (16.67, 19.73)	0.706
DBP (kPa)	10.67 ± 1.57	11 ± 1.53	0.143
ALT (U/L)	17 (10.25, 28.5)	24 (15, 39.75)	0.001
AST (U/L)	20 (16, 27.75)	22 (18, 29.75)	0.067
SUA (μmol/L)	314.5 (254, 401)	342 (271.75, 443)	0.074
Scr (μmol/L)	63.5 (52.5, 79.75)	62 (52, 77)	0.702
eGFR (mL/min/1.73 m2)	106.55 (87, 126.35)	116.95 (92.65, 139.78)	0.162
UACR (mg/g)	2.48 (0.97, 6.27)	2.11 (1.09, 7.82)	0.315
TC (mmol/L)	4.45 (3.62, 5.2)	4.8 (4.17, 5.6)	0.031
TG (mmol/L)	1.14 (0.94, 1.47)	1.81 (1.14, 2.61)	< 0.001
HDL (mmol/L)	1.18 (0.97, 1.3)	1.06 (0.91, 1.2)	0.012
LDL (mmol/L)	2.91 (2.3, 3.5)	3.25 (2.65, 3.91)	0.036
GLU0 min (mmol/L)	8.86 ± 2.48	9.11 ± 2.37	0.468
GLU120 min (mmol/L)	19.94 ± 5.71	19.75 ± 4.04	0.81
AUCGLU 0-180 min	3109.48 ± 804.22	3100.06 ± 590.39	0.931
HBA1c (%)	9.55 (7.23, 11.3)	8.9 (7.7, 10.73)	0.367
INS0 min (mIU/L)	0.59 (0.28, 0.76)	0.86 (0.53, 1.1)	< 0.001
INS120 min (mIU/L)	1.19 ± 0.41	1.41 ± 0.44	0.001
AUCINS 0-180 min	3.38 ± 0.4	3.6 ± 0.41	< 0.001
HOMA-IR	0.17 (-0.14, 0.35)	0.46 (0.13, 0.46)	< 0.001
CP0 min (μg/L)	1.37 (0.79, 2.04)	2.14 (1.37, 2.98)	< 0.001
CP120 min (μg/L)	3.75 (2.81, 5.47)	5.32 (3.54, 7.89)	0.001
AUCCP 0-180 min	569.48 (418.24, 745.2)	834.68 (521.36, 1155)	< 0.001
GCG0 min (pmol/L)	0.97 ± 0.21	1.11 ± 0.26	< 0.001
GCGpeak (pmol/L)	1.0 6± 0.2	1.2 ± 0.24	< 0.001
AUCGCG 0-180 min	3.1 ± 0.24	3.3 ± 0.26	< 0.001
LSM (kPa)	6 (4.08, 7)	7 (6, 8.33)	< 0.001
APRI	-0.59 ± 0.29	-0.51 ± 0.29	0.224
GPR	-0.65 ± 0.37	-0.52 ± 0.34	0.014
FIB4	0.18 ± 0.2	0.10 ± 0.27	0.016
PBF (%)	25.57 ± 8.46	30.64 ± 6.73	< 0.001
BMR (kcal)	1350.15 ± 173.24	1486.33 ± 204.92	< 0.001
VFA (cm2)	75.35 (57.18, 94.9)	98.05 (81.4, 146.15)	< 0.001
FFMI (kg/m2)	16.7 (14.93, 17.85)	18.05 (16.4, 19.48)	< 0.001
FMI (kg/m2)	5.85 (4.45, 7.23)	7.35 (6.13, 9.98)	< 0.001
SMI (kg/m2)	6.9 (5.85, 7.5)	7.5 (6.7, 8.08)	< 0.001

Data are mean ± SD or median (interquartile range). χ² test for gender comparison; The t-test was used for normally distributed variables; the Mann-Whitney U test for nonnormally distributed variables. BMI: Body mass index; WC: Waist circumference; WHR: Waist-to-hip ratio; SBP: Systolic blood pressure; DBP: Diastolic blood pressure; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; SUA: Serum uric acid; Scr: Serum creatinine; eGFR: Estimated glomerular filtration rate; UACR: Urinary albumin to creatinine ratio; TC: Total cholesterol; TG: Triglycerides; HDL: High-density lipoprotein; LDL: Low-density lipoprotein; GLU: Glucose; GCG: Glucagon; AUC: Area under the curve; HBA1c: Glycosylated hemoglobin; INS: Insulin; HOMA-IR: Homeostatic model assessment of insulin resistance; CP: C-peptide; LSM: Liver stiffness measurement; APRI: Aspartate aminotransferase to platelet ratio index; GPR: Gamma glutamyl transpeptidase to platelet ratio; FIB4: Fibrosis 4 score; PBF: Percent body fat; BMR: Basal metabolic rate; VFA: Visceral fat area; FFMI: Fat free mass index; FMI: Fat mass index; SMI: Skeletal muscle index.

Table 2 Correlation analysis between ultrasound attenuation parameter and other parameters

	r value	P value
Age	-0.331	< 0.001
Diabetes duration	-0.135	0.05
BMI	0.685	< 0.001
WC	0.513	< 0.001
SBP	0.039	0.573
DBP	0.203	0.003
ALT	0.205	0.003
AST	0.082	0.232
SUA	0.273	< 0.001
Scr	0.012	0.857
eGFR	0.046	0.505
UACR	0.031	0.649
TC	0.088	0.202
TG	0.27	< 0.001
HDL	-0.249	< 0.001
LDL	0.127	0.066
GLU0 min	0.067	0.329
GLU120 min	-0.055	0.43
AUCGLU 0-180 min	-0.027	0.698
HBA1c	-0.068	0.328
INS0 min1	0.361	< 0.001
INS120 min1	0.277	< 0.001
AUCINS 0-180 min1	0.289	< 0.001
HOMA-IR1	0.363	< 0.001
CP0 min	0.406	< 0.001
CP120 min	0.256	< 0.001
AUCCP 0-180 min	0.289	< 0.001
GCG0 min1	0.336	< 0.001
GCGpeak1	0.353	< 0.001
AUCGCG 0-180 min1	0.353	< 0.001
LSM	0.417	< 0.001
APRI1	0.102	0.137
GPR1	0.202	0.003
FIB41	-0.262	< 0.001
PBF	0.423	< 0.001
BMR	0.474	< 0.001
VFA	0.53	< 0.001
FFMI	0.509	< 0.001
FMI	0.536	< 0.001
SMI	0.476	< 0.001

¹Log-transformed variables.

Pearson correlation analysis for normally distributed variables; Spearman correlation analysis for nonnormally distributed variables. P values < 0.05 were considered significant. BMI: Body mass index; WC: Waist circumference; SBP: Systolic blood pressure; DBP: Diastolic blood pressure; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; SUA: Serum uric acid; Scr: Serum creatinine; eGFR: Estimated glomerular filtration rate; UACR: Urinary albumin to creatinine ratio; TC: Total cholesterol; TG: Triglycerides; HDL: High-density lipoprotein; LDL: Low-density lipoprotein; GLU: Glucose; GCG: Glucagon; AUC: Area under the curve; HBA1c: Glycosylated hemoglobin; INS: Insulin; HOMA-IR: Homeostatic model assessment of insulin resistance; CP: C-peptide; LSM: Liver stiffness measurement; APRI: Aspartate aminotransferase to platelet ratio index; GPR: Gamma glutamyl transpeptidase to platelet ratio; FIB4: Fibrosis 4 score; PBF: Percent body fat; BMR: Basal metabolic rate; VFA: Visceral fat area; FFMI: Fat free mass index; FMI: Fat mass index; SMI: Skeletal muscle index.

Table 3 Covariance analysis of ultrasound attenuation parameter-related factors

	Collinearity statistics
	Tolerance	VIF
Age	0.457	2.191
BMI	0.085	11.734
WC	0.367	2.726
DBP	0.770	1.298
ALT	0.580	1.723
SUA	0.698	1.432
TG	0.785	1.273
HDL	0.787	1.271
INS0 min	0.013	75.684
INS120 min	0.008	131.244
AUCINS 0-180 min	0.005	207.055
HOMA-IR	0.029	34.293
CP0 min	0.126	7.945
CP120 min	0.006	154.348
AUCCP 0-180 min	0.005	185.228
GCG0 min	0.156	6.404
GCGpeak	0.054	18.686
AUCGCG 0-180 min	0.075	13.308
LSM	0.607	1.647
GPR	0.423	2.363
FIB4	0.422	2.368
PBF	0.055	18.162
BMR	0.064	15.617
VFA	0.042	23.946
FFMI	0.060	16.617
FMI	0.024	42.368
SMI	0.029	34.632

Variance inflation factor > 10 or tolerance < 0.1 indicated a strong covariance between the independent variables. UAP: Ultrasound attenuation parameter; VIF: Variance inflation factor; BMI: Body mass index; WC: Waist circumference; DBP: Diastolic blood pressure; ALT: Alanine aminotransferase; SUA: Serum uric acid; TG: Triglycerides; HDL: High-density lipoprotein; GCG: Glucagon; AUC: Area under the curve; INS: Insulin; HOMA-IR: Homeostatic model assessment of insulin resistance; CP: C-peptide; LSM: Liver stiffness measurement; GPR: Gamma glutamyl transpeptidase to platelet ratio; FIB4: Fibrosis 4 score; PBF: Percent body fat; BMR: Basal metabolic rate; VFA: Visceral fat area; FFMI: Fat free mass index; FMI: Fat mass index; SMI: Skeletal muscle index.

Table 4 Multivariate linear regression analysis of various biomarkers with ultrasound attenuation parameter

	Standard β	t value	P value
Age	0.078	0.991	0.323
WC	-0.005	-0.067	0.947
DBP	-0.048	-0.756	0.451
ALT	0.093	1.258	0.210
SUA	0.034	0.512	0.609
TG	0.033	0.526	0.599
HDL	0.050	0.794	0.428
CP0 min	0.245	3.566	< 0.001
GCG0 min	0.182	2.020	0.045
LSM	0.320	5.144	< 0.001
GPR	-0.094	-1.088	0.278
FIB4	0.085	0.977	0.330

WC: Waist circumference; DBP: Diastolic blood pressure; ALT: Alanine aminotransferase; SUA: Serum uric acid; TG: Triglycerides; HDL: High-density lipoprotein; GCG: Glucagon; CP: C-peptide; LSM: Liver stiffness measurement; GPR: Gamma glutamyl transpeptidase to platelet ratio; FIB4: Fibrosis 4 score.

Table 5 Multivariate logistic regression model with metabolic dysfunction-associated steatotic liver disease as a dependent variable

	OR (95%CI)	P value
GCG0 min	3.068 (1.333-7.064)	0.008
CP0 min	1.965 (1.323-2.918)	0.001
LSM	1.422 (1.16-1.743)	0.001

This model was adjusted for the following covariates: Age, waist circumference, diastolic blood pressure, alanine aminotransferase, serum uric acid, triglycerides, high-density lipoprotein, gamma glutamyl transpeptidase to platelet ratio, fibrosis 4 score. Multivariate logistic regression analysis including GCG_{0 min} as a categorized binary variable using the cut-off value of 10.04 pmol/L. GCG_{0 min} was given value 0 in patients with GCG_{0 min} ≤ 10.04 pmol/L (n = 79) and value 1 in patients with GCG_{0 min} > 10.04 pmol/L (n = 133). Ultrasound attenuation parameter (UAP) was given value 0 in patients with UAP ≤ 244 dB/m (n = 68; without fatty liver) and value 1 in patients with UAP > 244 dB/m (n = 144; with fatty liver). GCG: Glucagon; CP: C-peptide; LSM: Liver stiffness measurement.