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

Johan W E Jocken, Dorien Reijnders, Emanuel E Canfora, Mark V Boekschoten, Joghum Plat, Gijs H Goossens, Ellen E Blaak, Johan W E Jocken, Dorien Reijnders, Emanuel E Canfora, Mark V Boekschoten, Joghum Plat, Gijs H Goossens, Ellen E Blaak

Abstract

The intestinal microbiota may contribute to the development of obesity by affecting host lipid metabolism and insulin sensitivity. To investigate the effects of microbiota manipulation on ex vivo basal and β-adrenergically-stimulated lipolysis in human adipocytes, 36 obese men were randomized to amoxicillin (broad-spectrum antibiotic), vancomycin (narrow-spectrum antibiotic) or placebo treatment (7 d, 1500 mg/d). Before and after treatment, ex vivo adipose tissue lipolysis was assessed under basal conditions and during stimulation with the non-selective β-agonist isoprenaline using freshly isolated mature adipocytes. Gene (targeted microarray) and protein expression were analyzed to investigate underlying pathways. Antibiotics treatment did not significantly affect basal and maximal isoprenaline-mediated glycerol release from adipocytes. Adipose tissue β-adrenoceptor expression or post-receptor signalling was also not different between groups. In conclusion, 7 d oral antibiotics treatment has no effect on ex vivo lipolysis in mature adipocytes derived from adipose tissue of obese insulin resistant men.

Trial registration: ClinicalTrials.gov NCT02241421.

Keywords: Adipose Tissue; Fatty acid metabolism; Insulin resistance; Lipolysis; Microbiota; Obesity.

Figures

Figure 1.
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.
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.
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.
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.

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