Pre-analytical sample handling standardization for reliable measurement of metabolites and lipids in LC-MS-based clinical research

Sens, Ashley; Rischke, Samuel; Hahnefeld, Lisa; Dorochow, Erika; Schäfer, S.M.G.; Thomas, Dominique; Köhm, Michaela; Geißlinger, Gerd; Behrens, Frank

doi:10.1016/j.jmsacl.2023.02.002

Cited by 24 publications

(18 citation statements)

References 56 publications

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“…Prostanoid levels were measured as previously described (Sens et al, 2023) with slight adaptions for the analysis of tissue samples. Briefly, tissues were homogenized by wet bead milling using zirconium oxide beads in 2‐ml reinforced tubes (Bertin Instruments, Frankfurt, Germany) and a Precellys 24‐Dual with a Cryolys cooling module (Bertin Instruments) as previously described (Kratz et al, 2022).…”

Section: Methodsmentioning

confidence: 99%

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Meloxicam treatment disrupts the regional structure of innate inflammation sites by targeting the pro‐inflammatory effects of prostanoids

Schäufele,

Kolbinger,

Friedel

et al. 2023

British J Pharmacology

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Background and PurposeNon‐steroidal anti‐inflammatory drugs (NSAIDs) are the most widely prescribed drugs in the world due to their analgesic, antipyretic and anti‐inflammatory effects. However, NSAIDS inhibit prostanoid synthesis, interfering with their pro‐inflammatory and anti‐inflammatory functions, and potentially prolonging acute inflammation.Experimental ApproachWe used high‐content immunohistochemistry to define the impact of meloxicam treatment on spatially separated pro‐ and anti‐inflammatory processes during innate inflammation in mice induced by zymosan. This allowed us to determine the effect of meloxicam treatment on the organization of pro‐ and anti‐inflammatory microenvironments thereby identifying relevant changes in immune cell localization, recruitment and activation.Key ResultsMeloxicam treatment reduced zymosan‐induced thermal hypersensitivity at early time points but delayed its resolution. High‐content immunohistochemistry revealed that the pro‐inflammatory area was smaller after treatment, diminishing neutrophil recruitment, M1‐like macrophage polarization, and especially phagocytosis by neutrophils and macrophages. The polarization of macrophages toward the M2‐like anti‐inflammatory phenotype was unaffected and the number of anti‐inflammatory eosinophils actually increased.Conclusion and ImplicationsHigh‐content immunohistochemistry was able to identify relevant meloxicam‐mediated effects on inflammatory processes based on alterations in the regional structure of inflammation sites. Meloxicam delayed the clearance of pathogens by inhibiting pro‐inflammatory processes, causing prolonged inflammation. Our data suggest that the prescription of NSAIDs as a treatment during an acute pathogen‐driven inflammation should be reconsidered in patients with compromised immune systems.

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Section: Methodsmentioning

confidence: 99%

“…Prostanoid levels were measured as previously described (Sens et al, 2023) with slight adaptions for the analysis of tissue samples.…”

Section: Prostanoid Measurementmentioning

confidence: 99%

Meloxicam treatment disrupts the regional structure of innate inflammation sites by targeting the pro‐inflammatory effects of prostanoids

Schäufele,

Kolbinger,

Friedel

et al. 2023

British J Pharmacology

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“…Bioactive lipids including sphingolipids and ceramides, lysophosphatidic acids, endocannabinoids, eicosanoids (oxylipins) and pterins were analyzed in plasma by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) as described in detail in the supplementary material of a previous study [8]. Analytical methods were optimized based on methods reported in our previous studies [9,[51][52][53][54].…”

Section: Targeted Analyses Of Lipid Signaling Moleculesmentioning

confidence: 99%

Sex, age, body mass index, and contraceptive use but not perceived stress influence healthy plasma lipidomic profiles

Hahnefeld,

Hackel,

Trautmann

et al. 2024

Preprint

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Background The stress of everyday life is thought to contribute to the pathogenesis of metabolic, vascular, mental, and immune diseases, with different susceptibilities in women and men. We hypothesized that sex differences in stress perception may manifest in sex-dependent lipid profiles. Methods The present study investigated the effect of sex, age, body mass index, perceived stress, regular and occasional drug use, and dietary supplements on plasma lipidomic profiles, obtained by mass spectrometry analyses. The study included 217 healthy women and 108 healthy men aged 18–68 years, who were recruited in a 2:1 female:male ratio to account for women with/without contraceptives. Results As expected, dehydroepiandrosterone sulfate (DHEAS) and ceramides were higher in men than in women, and in both sexes DHEAS decreased with age, while ceramides increased. Contrary to expectations, neither DHEAS nor ceramides were associated with perceived stress (PSQ30 questionnaire), which peaked in young and 51 + women (low in ages between), and in obese young men, whereas the overall male PSQ30 peak was around 40 years of age. None of the lipid species or classes showed a similar "age X sex X BMI" interaction. Strong sex differences were found for lysophosphatidylcholines (LPCs) (low in women) and their metabolites, lysophosphatidic acids (LPAs) (high in women). The LPA:LPC ratio was particularly high in women receiving contraceptives suggesting a strong hormone-induced conversion of LPCs to LPAs via autotaxin, which was much higher in women than in men, and is known to trigger platelet aggregation. In addition, phosphatidylethanolamines (PE) were high in women. They are precursors of endocannabinoids such as palmitoylethanolamide (PEA), which was elevated in subjects with a medical history of hypertension and increased with the BMI. Conclusion The results reveal complex sex differences in perceived stress and lipidomic profiles, the latter being exacerbated by contraceptive use, but perceived stress and lipids were not directly correlated. Trial registration Not applicable

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“…The analysis of endocannabinoids was performed by adapting a method that was originally developed for the analysis of plasma samples. Briefly, cell pellets were resuspended in 200 µL of water and extracted using liquid-liquid-extraction as previously described in detail [83].…”

Section: Liquid Chromatography-tandem Mass Spectrometrymentioning

confidence: 99%

Sphingosine Kinases at the Intersection of Pro-Inflammatory LPS and Anti-Inflammatory Endocannabinoid Signaling in BV2 Mouse Microglia Cells

Standoli

Rapino

Meo

et al. 2023

IJMS

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Microglia, the resident immune cells of the central nervous system, play important roles in brain homeostasis as well as in neuroinflammation, neurodegeneration, neurovascular diseases, and traumatic brain injury. In this context, components of the endocannabinoid (eCB) system have been shown to shift microglia towards an anti-inflammatory activation state. Instead, much less is known about the functional role of the sphingosine kinase (SphK)/sphingosine-1-phosphate (S1P) system in microglia biology. In the present study, we addressed potential crosstalk of the eCB and the S1P systems in BV2 mouse microglia cells challenged with lipopolysaccharide (LPS). We show that URB597, the selective inhibitor of fatty acid amide hydrolase (FAAH)—the main degradative enzyme of the eCB anandamide—prevented LPS-induced production of tumor necrosis factor-α (TNFα) and interleukin-1β (IL-1β), and caused the accumulation of anandamide itself and eCB-like molecules such as oleic acid and cis-vaccenic acid ethanolamide, palmitoylethanolamide, and docosahexaenoyl ethanolamide. Furthermore, treatment with JWH133, a selective agonist of the eCB-binding cannabinoid 2 (CB2) receptor, mimicked the anti-inflammatory effects of URB597. Interestingly, LPS induced transcription of both SphK1 and SphK2, and the selective inhibitors of SphK1 (SLP7111228) and SphK2 (SLM6031434) strongly reduced LPS-induced TNFα and IL-1β production. Thus, the two SphKs were pro-inflammatory in BV2 cells in a non-redundant manner. Most importantly, the inhibition of FAAH by URB597, as well as the activation of CB2 by JWH133, prevented LPS-stimulated transcription of SphK1 and SphK2. These results present SphK1 and SphK2 at the intersection of pro-inflammatory LPS and anti-inflammatory eCB signaling, and suggest the further development of inhibitors of FAAH or SphKs for the treatment of neuroinflammatory diseases.

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Pre-analytical sample handling standardization for reliable measurement of metabolites and lipids in LC-MS-based clinical research

Cited by 24 publications

References 56 publications

Meloxicam treatment disrupts the regional structure of innate inflammation sites by targeting the pro‐inflammatory effects of prostanoids

Meloxicam treatment disrupts the regional structure of innate inflammation sites by targeting the pro‐inflammatory effects of prostanoids

Sex, age, body mass index, and contraceptive use but not perceived stress influence healthy plasma lipidomic profiles

Sphingosine Kinases at the Intersection of Pro-Inflammatory LPS and Anti-Inflammatory Endocannabinoid Signaling in BV2 Mouse Microglia Cells

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