Potential role of anti-inflammatory HDL subclasses in metabolic unhealth/obesity

Figures & data

Table 1. Characteristics of the study cohort at baseline.

Variable	MHNW, control n = 154	MHO n = 38	MUNW n = 51	MUO n = 73	p Value
Age (years)	44 (37–51)	48 (42–54)^b,c	59 (50–65)^a	63 (55–71)^a	.00
Sex male, n(%)	41 (26.6)	13 (34.2)	15 (29.4)	28 (38.4)	.324
Current smoking, n(%)	19 (12.8)	2 (5.4)	3 (6.0)	9 (12.9)	.677
Current alcohol consumption, n(%)	27 (17.5)	13 (34.2)	6 (11.8)	18 (24.7)	.689
Exercise ≥30 min/week, (%)	86 (55.8)	17 (44.7)	33 (64.7)	58 (79.4)	.135
SBP (mmHg)	113 (108–120)	117 (112–122)^b,c	135 (127–153)^a,b	146 (128–155)^a	.000
DBP (mmHg)	75 (70–79)	78 (66–79)^b,c	84 (78–90)^a,b	88 (78–95)^a	.000
FPG (mmol/L)	4.7 (4.4–4.9)	4.9 (4.5–5.2)^b,c	6.2 (5.7–7.0)^a,b	6.7 (5.9–7.5)^a	.000
TG (mmol/L)	0.9 (0.7–1.1)	1.0 (0.8–1.3)^b,c	1.6 (1.0–2.3)^a	1.8 (1.4–2.5)^a	.000
HDL-C (mmol/L)					.000
Male	1.5 ± 0.05	1.5 ± 0.06	1.4 ± 0.08	1.2 ± 0.03^a,b
Female	1.7 ± 0.03	1.7 ± 0.03	1.6 ± 0.07	1.4 ± 0.04^a,b
TC (mmol/L)	5.0 ± 1.0	5.5 ± 0.9	5.9 ± 1.3^a	5.6 ± 1.1^a	.000
LDL-C (mmol/L)	2.8 (2.3–3.3)	3.3 (2.6–3.5)	3.4 (2.6–-4.2)^a	3.2 (2.6–3.9)^a	.012
VLDL-C (mmol/L)	0.4 (0.3–0.5)	0.4 (0.3–0.5)^b,c	0.7 (0.4–1.0)^a	0.8 (0.6–1.1)^a	.000
Apo B (mg/dL)	0.8 ± 0.2	1.0 ± 0.2	1.0 ± 0.2^a	1.0 ± 0.2^a	.000
BMI (kg/m²)	21.5 (19.8–22.7)	25.6(25.1–28.2)^a,c	22.8 (20.2–23.8)^a,b	27.2 (26.2-–29.1)^a	.000
Waist circumference (cm)
Male	77.1 ± 5.7	94.5 ± 3.8^a,c	84.1 ± 7.1^a,b	96.7 ± 5.1^a	.003
Female	73.7 ± 5.2	88.6 ± 9.4^a,c	78.1 ± 5.1^a,b	94.5.6^a	.000
Fasting insulin (mmol/L)	7.8 (5.5–10.3)	13.3 (7.9–17.9)^a	9.6 (6.1–13.4)^b	13.1 (8.9–7.4)^a	.000
HOMA-IR	1.5 (1.1–2.1)	3.1 (1.7–3.8)^a,b	2.9 (1.6–3.7)^a,b	3.8 (2.8–5.7)^a	.000

Data are the mean ± SE, or median (25% quartile, 75% quartile).

SBP: systolic blood pressure;DBP: diastolic blood pressure; BMI: body mass index; TC: total cholesterol; HDL-C: high-density lipoprotein-cholesterol; LDL-C: low-density lipoprotein-cholesterol; TG: triglycerides; HOMA-IR: Homoeostatic Model Assessment of Insulin Resistance; hs-CRP: high-sensitivity C-reactive protein; IL-6: interleukin-6.

^ap < .05 vs. control group.

^bp < .05 vs. MUO group.

^cp < .05 vs. MUNW group.

^cp < .05 vs. MUNW group.

Figure 1. Distribution of HDL subclasses between groups. Level of HDL2b-c (%) (A), HDL3-c (%) (B), HDL2b-c (mmol/L) (C) and HDL3-c (mmol/L) (D) in the four groups. The bar height indicates the proportion of each HDL subclasses expressed as a percentage relative to total HDL-protein as determined by absorbance ± standard error. *p < .05, **p < .01 by one-way ANOVA.

Figure 2. Alteration of HDL related proteins between groups. Level of apoA-1 (mg/dL) (A), RBP (mg/L) (B), PON-1 (ng/ml) (C) in the four groups. The bar height indicates the proportion of each protein expressed as a percentage relative to total value as determined by absorbance ± standard error. *p < .05, **p < .01 by one-way ANOVA.

Figure 3. Inflammatory cells counts and inflammation factors in the four groups. White blood cells count (A), neutrophils count (B), monocyte count (C) and lymphocytes count (D) in controls, MHO, MUNW and MUO. The levels of IL-6 (D) and hs-CRP (E) in controls, MHO, MUNW and MUO. *p < .05, **p < .01 by one-way ANOVA.

Table 2. Correlations between HDL subclasses and metabolic factors and inflammatory indexes.

	MHNW		MHO		MUNW		MUO
	HDL2b	HDL3	HDL2b	HDL3	HDL2b	HDL3	HDL2b	HDL3
waist	−0.37*	0.20	−0.60**	0.52**	−0.26	0.27	−0.25*	0.18
SBP	0.02	−0.04	0.36*	−0.26*	0.35*	−0.32*	−0.02	0.20
TG	−0.27*	0.30**	−0.51**	0.47**	−0.47**	0.40**	−0.45**	0.33**
HDL-C	0.55**	−0.56**	0.76**	−0.64**	0.73**	−0.51**	0.71**	−0.40**
FPG	−0.21*	0.078	0.23	−0.32	−0.14	0.17	−0.24*	0.28*
IR index	−0.21	0.06	−0.51**	0.36**	−0.44**	0.35**	−0.11*	0.19*
RBP	−0.27*	0.24	−0.38**	0.35	−0.35**	0.25	−0.35**	0.18
apoA1	0.15*	−0.13*	0.56**	−0.40**	0.45**	−0.32*	0.59**	−0.31**
PON-1	0.16	−0.30	−0.40*	0.40	0.09*	0.15	−0.02*	0.15
WBC	−0.11	0.20	−0.40**	0.30	−0.36**	0.37**	−0.13	0.11
lymphocyte	−0.31**	0.32	−0.23**	0.11	−0.30**	0.32*	−0.20*	0.26*
monocyte	−0.05	0.05	−0.32	0.23	−0.22	0.37*	−0.12	0.30
IL-6	−0.07	0.22	−0.16	0.13	−0.06	0.05	−0.10	−0.10
hs-CRP	−0.31**	0.22	−0.40**	0.26	−0.43**	0.14**	−0.31**	0.13*

Univariate correlations of serum HDL subclasses with waist, characteristics of metabolic syndrome including SBP, TG, HDL-Cand FPG serum lipid, IR index, HDL related proteins (PON-1, apoA-1 and ), inflammatory cells (WBC and lymphocyte) and hs-CRP in 154 controls, in 38 subjects with metabolic healthy overweight or obesity, in 51 subjects with metabolic unhealthy normal weight and in 73 with metabolic unhealthy overweight or obesity.

Spearman correlation coefficients were shown. SBP: systolic blood pressure; TG: triglycerides; HDL-C: high-density lipoprotein cholesterol; FPG: fasting plasma glucose; IR: Insulin Resistance index; RBP: retinol-binding protein; PON-1: Paraoxonase-1; WBC: white blood cell; IL-6: interleukin-6; hs-CRP: high-sensitivity C-reactive protein.

*p < .05.

**p < .01.

Table 3. Stepwise linear regression models.

Independent variable	Standardized coefficient β (SE)	p Value
A Stepwise linear regression model for percentage of HDL2b-c (%) (n = 316).
PON1	0.030	.006
Log TG	−0.421 (1.984)	.000
Log ApoA1	0.303 (3.605)	.000
Log hs-CRP	−0.226 (1.040)	.000
Log HOMA-IR	−0.166 (1.526)	.011
Total R²		.000
B Stepwise linear regression model for percent of HDL3-c (%) (n = 316).
Log TG	0.369 (2.183)	.000
Log ApoA1	−0.224 (3.915)	.000
Log HOMA-IR	0.199 (1.580)	.002
Total R²		.000

(A) stepwise method was employed among levels of PON1, TG, apoA1, hs-CRP and IL-6, and other known risk factors of HDL2b-c, including age, sex, smoking status, physical exercise, BMI and HOMA-IR. Only variables remaining in the final model are presented.

(B) stepwise method was employed among levels of hs-CRP, TG, IL-6 and other known risk factors of HDL3-c, including age, sex, smoking status, physical exercise, BMI and HOMA-IR. Only variables remaining in the final model are presented.

Table 4. Multivariable linear regression analysis of HDL subclasses and inflammatory indexes.

Indices	Multivariable
	Unstandardized Coefficient β (95% CI)	p Value
WBC
HDL2b (log per 1 %)	−4.80 (−8.13− −1.46)	.005
HDL3 (log per 1 %)	3.80 (1.50–6.10)	.001
Lymphocytes
HDL2b (log per 1 %)	−1.53 (−2.54− −0.41)	.008
HDL3 (log per 1 %)	1.12 (0.36–1.89)	.004
hs-CRP
HDL2b (log per 1 %)	−9.67 (−14.43− −4.91)	.000
HDL3 (log per 1 %)	1.89 (−1.49−5.27)	.27

Association of HDL subclasses with different inflammation cells and factor using multivariable linear regression analysis in subjects with MUO.

Values are expressed as unstandardized coefficient β and 95% confidence interval (CI). Abbreviations are the same as in Table 1. WBC: white blood cell. Covariates in the multivariable model included age and sex.

Figure 4. The result of multinomial logistic regression analysis. Significant results of multinomial logistic regression analysis to assess the association between HDL subclasses and metabolic syndrome/obesity adjusted by sex, age, moderate physical activity, and quitting smoking. *p < .05. **p < .01. Metabolically healthy normal weight (MHNW) was set as the reference group.