Langkilde Anne, Andersen Ove, Henriksen Jens H, Langberg Henning, Petersen Janne, Eugen-Olsen Jesper
Clinical Research Centre, Copenhagen University Hospital, Hvidovre, Denmark.
Clin Physiol Funct Imaging. 2015 Mar;35(2):110-9. doi: 10.1111/cpf.12134. Epub 2014 Feb 4.
Inflammation, and specifically adipose tissue (AT) inflammation, is part of the pathophysiology of obesity and HIV-associated lipodystrophy. Local AT protein assessment methods are limited, and AT inflammation studies have therefore primarily examined inflammatory gene expression. We therefore investigated the utility of microdialysis to study in situ AT interstitial inflammatory protein levels.
Abdominal subcutaneous AT microdialysis was performed in six healthy men, six HIV-infected men with lipodystrophy and six without lipodystrophy using the internal references (51) Cr-EDTA and (125) I-human serum albumin. We measured 41 inflammatory proteins in microdialysis samples by Luminex technology, as well as systemic levels in 14 subjects. Furthermore, in vitro studies of the internal reference technique for microdialysis recovery of inflammatory proteins were made.
We detected in situ AT interstitial levels of 14 inflammatory proteins by microdialysis, while the 27 other inflammatory proteins assessed were only detected sporadically. Initial levels of IL-6 and IL-8 were undetectable. Insertion trauma affected IL-1α, IL-6, IL-8, monocyte chemotactic factor (MCP)-1, IP-10, G-CSF, growth-related oncogene (GRO), macrophage-derived chemokine (MDC) and macrophage inflammatory protein (MIP)-1β levels, while fibroblast growth factor (FGF)-2 was not affected. Systemic and AT interstitial levels were poorly correlated. The microdialysis recovery of smaller proteins was higher than for larger, and the internal references improved microdialysis by accounting for variation in perfusion across the membrane.
Interstitial inflammatory proteins can be sampled in situ using microdialysis. Use of internal references improves the microdialysis technique. However, insertion trauma hampers the use of microdialysis to study AT inflammatory levels, except for FGF-2. Still, microdialysis gives unique insight to in situ AT interstitial concentrations.
炎症,尤其是脂肪组织(AT)炎症,是肥胖症和HIV相关脂肪代谢障碍病理生理学的一部分。局部AT蛋白质评估方法有限,因此AT炎症研究主要检测炎症基因表达。因此,我们研究了微透析用于原位研究AT间质炎症蛋白水平的效用。
对6名健康男性、6名患有脂肪代谢障碍的HIV感染男性和6名未患脂肪代谢障碍的HIV感染男性进行腹部皮下AT微透析,使用内部参考物(51)Cr - EDTA和(125)I - 人血清白蛋白。我们通过Luminex技术测量微透析样本中的41种炎症蛋白,以及14名受试者的全身水平。此外,还对炎症蛋白微透析回收率的内部参考技术进行了体外研究。
通过微透析检测到14种炎症蛋白的原位AT间质水平,而评估的其他27种炎症蛋白仅偶尔检测到。IL - 6和IL - 8的初始水平无法检测到。插入创伤影响IL - 1α、IL - 6、IL - 8、单核细胞趋化因子(MCP)-1、IP - 10、G - CSF、生长相关癌基因(GRO)、巨噬细胞衍生趋化因子(MDC)和巨噬细胞炎性蛋白(MIP)-1β水平,而成纤维细胞生长因子(FGF)-2不受影响。全身和AT间质水平相关性较差。较小蛋白质的微透析回收率高于较大蛋白质,内部参考物通过考虑跨膜灌注的变化改进了微透析。
可使用微透析原位采集间质炎症蛋白。使用内部参考物改进了微透析技术。然而,除FGF - 2外,插入创伤妨碍了使用微透析研究AT炎症水平。尽管如此,微透析为原位AT间质浓度提供了独特的见解。