Effects of gut microbiota manipulation on ex vivo lipolysis in human abdominal subcutaneous adipocytes

Figures & data

Table 1. Baseline Characteristics.

	PLA (N = 7)	AMOX (N = 7)	VANCO (N = 8)	P-value
Age (years)	60.6 ± 3.3	53.3 ± 2.2	57.5 ± 2.9	0.231
Body Weight (kg)	97.2 ± 3.7	98.2 ± 2.7	97.3 ± 3.6	0.973
BMI (kg/m^2)	31.9 ± 0.9	31.6 ± 0.7	31.4 ± 0.8	0.916
Fasting glucose (mmol/l)	5.9 ± 0.2	5.9 ± 0.3	6.0 ± 0.2	0.942
Fasting Insulin (mU/l)	14.1 ± 2.8	20.4 ±3.0	19.2 ± 1.4	0.184
HOMA-IR	3.7 ± 0.7	5.4 ± 0.9	5.1 ± 0.3	0.206
TAG (mmol/l)	1.1 ± 0.2	1.6 ± 0.5	1.0 ± 0.2	0.475
FFA( umol/l)	587.9 ± 33.7	579.2 ± 53.8	593.0 ± 53.2	0.979

Characteristics given as mean ± SEM. The P-value represents the difference between the 3 groups (one-way ANOVA). BMI: Body mass index; HOMA-IR: Homeostasis model assessment for insulin resistance.

Table 2. Basal and maximal ISO- mediated lipolytic response in SCAT adipocytes before and after intervention, compared to placebo.

		PLA (N = 7)	AMOX (N = 7)	VANCO (N = 8)	P-value
Basal	pre	17840±4496	17879±4145	18871±4744	0.757
(μM/10^−6cells/3h)	post	19501±3740	14280±2543	15275±2735
Max	pre	64071±11535	88278±22031	67477±16633	0.591
(μM/10^−6cells/3h)	post	75306±14030	74682±24914	49357±9091
Halfway	pre	39847±6644	52751±11921	42544±10283	0.602
(μM/10^−6cells/3h)	post	45755±7449	43880±13147	32027±4301
EC50	pre	3.96E-07±1.48E-07	1.05E-06±3.22E-07	2.38E-06±5.72E-07	0.124
	post	2.40E-07±9.59E-08	2.22E-05±1.47E-05	1.00E-06±5.87E-07
pD2	pre	6.52±0.25	6.24±0.27	6.02±0.33	0.057
	post	7.06±0.32	5.77±0.51	6.76±0.43

Values are given as mean ± standard error of the mean (SEM). P-value represents the overall time × treat P-value of the repeated measures ANOVA (n = 22).

Figure 1. Dose-response curves for ISO-mediated lipolytic response in human mature adipocytes derived from the SCAT before and after intervention. Lipolysis (glycerol release in the medium) is expressed compared to baseline, following incubation with increasing concentrations ISO (10⁻¹⁰ to 10⁻⁴ mol/l) before (circles) and after (triangles) 7 d treatment with placebo (panel A), AMOX (B) or VANCO (C), n = 22.

Figure 2. Gene expression profiling of lipolysis-related genes in adipose tissue before and after intervention, compared to placebo. This heat map depicts fold changes (FC) observed after compared to before AMOX (A), VANCO (V) and PLA (P) intervention. Data are derived from n = 15 individuals.

Figure 3. Quantitative analysis of the Western blots of HSL (A), ATGL (B) and phosphorylated HSL on Ser563 (corresponding to human Ser552) (C). Pre (white bars) and post (black bars) intervention data are normalized for the loading control β-actin. Values are given as mean ± SEM (n = 5 for PLA, n = 5 for VANCO and n = 6 for AMOX).

Figure 4. Representative Western Blot for lipolytic markers in human adipose tissue. Membranes were probed with antibodies directed against total ATGL, total HSL, phosphorylated HSL (pHSL) on Ser563 (corresponding to human Ser552) and β-actin was used as a loading control. A subset of 3 subjects per group is shown.