The use of microdialysis to characterize the endocrine production of human subcutaneous adipose tissue in vivo

Dostálová, I; Kaválková, Petra; Haluzíková, D; Housová, J; Matoulek, Martin; Haluzı́k, Martin

doi:10.1016/j.regpep.2009.03.008

Cited by 13 publications

(8 citation statements)

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“…To circumvent this problem, osmotic agents such as Dextran-60 (~ 60 kDa) or -70 (~70 kDa) were being added at high concentrations (1 to 4% w/v). These agents have since been widely used in microdialysis sampling with 100 kDa MWCO membranes (Abrahamsson et al, 2012; Dabrosin, 2003; Dostalova et al, 2009; Dostálová et al, 2003; Farnebo et al, 2009; Förster et al, 2013; Hillman et al, 2005b; Hillman et al, 2006; Mellergard et al, 2008; Mellergård et al, 2011; Rosenbloom et al, 2006; Sjoegren and Anderson, 2009; Sjögren et al, 2012; Sjögren et al, 2002; von Grote et al, 2011; Waelgaard et al, 2008; Zhong et al, 2007). Our results show that Dextran-70, when allowed to enter the extracellular space, causes an intense inflammatory response.…”

Section: 0 Discussionmentioning

confidence: 99%

Comparison of microdialysis sampling perfusion fluid components on the foreign body reaction in rat subcutaneous tissue

Keeler¹,

Durdik²,

Stenken³

2014

European Journal of Pharmaceutical Sciences

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Microdialysis sampling is a commonly used technique for collecting solutes from the extracellular space of tissues in laboratory animals and humans. Large molecular weight solutes can be collected using high molecular weight cutoff (MWCO) membranes (100 kDa or greater). High MWCO membranes require addition of high molecular weight dextrans or albumin to the perfusion fluid to prevent fluid loss via ultrafiltration. While these perfusion fluid additives are commonly used during microdialysis sampling, the tissue response to the loss of these compounds across the membrane is poorly understood. Tissue reactions to implanted microdialysis sampling probes containing different microdialysis perfusion fluids were compared over a 7-day time period in rats. The base perfusion fluid was Ringer’s solution supplemented with either bovine serum albumin (BSA), rat serum albumin (RSA), Dextran-70, or Dextran-500. A significant inflammatory response to Dextran-70 was observed. No differences in the tissue response between BSA and RSA were observed. Among these agents, the BSA, RSA, and Dextran-500 produced a significantly reduced inflammatory response compared to the Dextran-70. This work demonstrates that use of Dextran-70 in microdialysis sampling perfusion fluids should be eliminated and replaced with Dextran-500 or other alternatives.

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Section: 0 Discussionmentioning

confidence: 99%

Comparison of microdialysis sampling perfusion fluid components on the foreign body reaction in rat subcutaneous tissue

Keeler¹,

Durdik²,

Stenken³

2014

European Journal of Pharmaceutical Sciences

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“…Microdialysis is a validated method to gain access to and sample adipose-derived molecules from the intercellular space, providing valuable information at a tissue level (5,20,27). These studies have indicated a risk of an artifact due to trauma from the catheter.…”

Section: Discussionmentioning

confidence: 99%

Systemic inflammation activates the nuclear factor-κB regulatory pathway in adipose tissue

Ekström

Halle²,

Bjessmo

et al. 2010

American Journal of Physiology-Endocrinology and Metabolism

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Adipose tissue (AT) is a store of energy but also an endocrine organ with the capacity to produce and release proinflammatory mediators into the circulation. The mechanism that may trigger human AT inflammation on a cellular level still remains largely unknown. The aim of this study was to investigate whether an acute systemic inflammation increases AT inflammatory activity, focused on innate immunity. Open heart surgery results in an extensive acute systemic inflammation. Therefore, we investigated the in vivo gene expression and production of inflammatory mediators in omental and subcutaneous AT stimulated by surgery. Biopsies from omental and subcutaneous AT were collected before and after cardiopulmonary bypass. Blood samples were collected at the same time as the AT biopsies and plasma IL-6 levels were measured with ELISA. RT-PCR was used for quantification of relative AT gene expression. To verify the gene expression results on a protein level, we used immunohistochemistry and microdialysis. After surgery, in both omental and subcutaneous AT, there was a strong upregulation of nuclear factor-kappaB-regulated genes, e.g., chemokine ligand-2, E-selectin, IL-1beta, IL-6, IL-8, and Toll-like receptor-2. Immunohistochemistry showed staining for E-selectin associated with a high number of macrophages in close contact with and in the vascular wall. Increased levels of IL-6 were detected in microdialysate from subcutaneous AT. In conclusion, we present the novel finding that this model of inflammation induced a strong inflammatory response in both omental and subcutaneous AT including adhesion of macrophages to an activated endothelium and release of IL-6 from AT interstitium. It can be hypothesized that AT exerts a modulatory effect on innate immunity in humans.

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“…Based on our in vitro data, we speculate that mutation carriers have a 50% reduction in resistin levels in adipose tissue, although this is not reflected by measured plasma levels. Several studies have, however, already indicated that resistin blood levels do not correlate to resistin adipose tissue levels (39)(40)(41). Therefore, we suspect that a reduction in resistin levels may be present in the adipose tissue, having a local effect on adipogenesis.…”

Section: Discussionmentioning

confidence: 81%

Identification and functional characterization of a missense mutation in resistin in two patients with severe obesity and insulin resistance

Beckers

Freitas²,

Zegers³

et al. 2011

European Journal of Endocrinology

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Objective: In this study, we hypothesized that mutations in the resistin encoding gene, RETN, may cause a monogenic form of obesity. Design/methods: We screened the coding region of RETN in 81 morbidly obese adults, 263 overweight and obese children/adolescents, and 116 healthy lean subjects. In vitro experiments include qPCR, ELISA, and western blot for WT and mutant resistin transfected into 3T3-L1 adipocytes. Results: Mutation analysis identified five sequence variants in our patient populations:0 -UTR C100 A/G, T73T, IV3-61 C/A, and C78S. In our control population, we only found the 3 0 -UTR C87 G/A variant. We started functional experiments for the C78S mutation that was found in a 20-year-old obese male (body mass index (BMI)Z39.7 kg/m 2 ) and his obese mother (BMIZ31.9 kg/m 2 ). In vitro testing demonstrated that the mutation does not impair mRNA expression. We identified a 100-fold lower extracellular protein concentration for mutant resistin compared with WT levels using a resistin ELISA on cell culture medium (PZ4.87!10 K6 ). We also detected a decreased intracellular concentration for the mutant protein (tenfold lower relative levels, PZ0.007). The plasma resistin levels of the proband and his mother, however, did not differ significantly from lean control individuals. Conclusions: In conclusion, we identified the first missense mutation in resistin in a morbidly obese proband and his obese mother. Functional testing of the mutant protein suggests that the C78S mutant protein is degraded, possibly resulting in a decreased extracellular concentration, which may predispose to obesity. 164 927-936 European Journal of Endocrinology

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The use of microdialysis to characterize the endocrine production of human subcutaneous adipose tissue in vivo

Cited by 13 publications

References 50 publications

Comparison of microdialysis sampling perfusion fluid components on the foreign body reaction in rat subcutaneous tissue

Comparison of microdialysis sampling perfusion fluid components on the foreign body reaction in rat subcutaneous tissue

Systemic inflammation activates the nuclear factor-κB regulatory pathway in adipose tissue

Identification and functional characterization of a missense mutation in resistin in two patients with severe obesity and insulin resistance

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