Development and validation of a rapid, specific and sensitive LC-MS/MS bioanalytical method for eicosanoid quantification - assessment of arachidonic acid metabolic pathway activity in hypertensive rats

Kij, Agnieszka; Kuś, Kamil; Czyżyńska-Cichoń, Izabela; Chłopicki, Stefan; Walczak, Maria

doi:10.1016/j.biochi.2020.03.010

Cited by 15 publications

(20 citation statements)

References 42 publications

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“…Analytes were separated under the gradient elution mode. The MS detection of studied eicosanoid and their deuterated internal standards was carried out in negative ion electrospray ionisation by applying the multiple reaction monitoring mode (MRM); this method and the instrument operation parameters were described previously [ 20 ].…”

Section: Methodsmentioning

confidence: 99%

Exogenous Vitamins K Exert Anti-Inflammatory Effects Dissociated from Their Role as Substrates for Synthesis of Endogenous MK-4 in Murine Macrophages Cell Line

Kieronska‐Rudek

Kij

Kaczara

et al. 2021

Cells

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Vitamins K exert a range of activities that extend far beyond coagulation and include anti-inflammatory effects, but the mechanisms involved in anti-inflammatory action remain unclear. In the present study, we showed that various forms of exogenous vitamins—K1, K3, K2 (MK-4, MK-5, MK-6 and MK-7)—regulated a wide scope of inflammatory pathways in murine macrophages in vitro, including NOS-2, COX-2, cytokines and MMPs. Moreover, we demonstrated for the first time that macrophages are able to synthesise endogenous MK-4 on their own. Vitamins with shorter isoprenoid chains—K1, K3 and MK-5—exhibited stronger anti-inflammatory potential than vitamins with longer isoprenoid chains (MK-6 and MK-7) and simultaneously were preferably used as a substrate for MK-4 endogenous production. Most interesting, atorvastatin pretreatment inhibited endogenous MK-4 production but had no impact on the anti-inflammatory activity of vitamin K. In summary, our results demonstrate that macrophages are able to synthesise endogenous MK-4 using exogenous vitamin K, and statin inhibits this process. However, the anti-inflammatory effect of exogenous vitamin K was independent of endogenous MK-4 synthesis.

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Section: Methodsmentioning

confidence: 99%

Exogenous Vitamins K Exert Anti-Inflammatory Effects Dissociated from Their Role as Substrates for Synthesis of Endogenous MK-4 in Murine Macrophages Cell Line

Kieronska‐Rudek

Kij

Kaczara

et al. 2021

Cells

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“…Selected eicosanoids (5-, 12-, 15-, and 20-HETE, 8,9-, 11,12-, and 14,15-EET, 8,9-, 11,12-, and 14,15-DHET) were quantified in plasma, Xyzyk-derived plasma, and Krebs-Hepes buffer collected after aorta incubation using a UPLC-MS/MS technique with the application of an already published methodology [53]. In short, each sample was spiked with a mixture of internal standards and gently mixed.…”

Section: Uplc-ms/ms Eicosanoid Analysismentioning

confidence: 99%

Thrombin Inhibition Prevents Endothelial Dysfunction and Reverses 20-HETE Overproduction without Affecting Blood Pressure in Angiotensin II-Induced Hypertension in Mice

Kij

Bar

Przyborowski

et al. 2021

IJMS

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Angiotensin II (Ang II) induces hypertension and endothelial dysfunction, but the involvement of thrombin in these responses is not clear. Here, we assessed the effects of the inhibition of thrombin activity by dabigatran on Ang II-induced hypertension and endothelial dysfunction in mice with a particular focus on NO- and 20-HETE-dependent pathways. As expected, dabigatran administration significantly delayed thrombin generation (CAT assay) in Ang II-treated hypertensive mice, and interestingly, it prevented endothelial dysfunction development, but it did not affect elevated blood pressure nor excessive aortic wall thickening. Dabigatran’s effects on endothelial function in Ang II-treated mice were evidenced by improved NO-dependent relaxation in the aorta in response to acetylcholine in vivo (MRI measurements) and increased systemic NO bioavailability (NO2− quantification) with a concomitant increased ex vivo production of endothelium-derived NO (EPR analysis). Dabigatran treatment also contributed to the reduction in the endothelial expression of pro-inflammatory vWF and ICAM-1. Interestingly, the fall in systemic NO bioavailability in Ang II-treated mice was associated with increased 20-HETE concentration in plasma (UPLC-MS/MS analysis), which was normalised by dabigatran treatment. Taking together, the inhibition of thrombin activity in Ang II-induced hypertension in mice improves the NO-dependent function of vascular endothelium and normalises the 20-HETE-depedent pathway without affecting the blood pressure and vascular remodelling.

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“…Plasma and urine samples were prepared for the LC-MS/MS assay according to the protocol slightly changed from that previously described in [22,23]. Briefly, after the addition of internal standard mixture, all samples were purified applying liquid-liquid extraction using acidified ethyl acetate.…”

Section: Measurement Of Eicosanoid Production By Lc-ms/msmentioning

confidence: 99%

“…Calibration and quality control samples were prepared using artificial plasma or urine [22,23] spiked with known concentration of eicosanoid working standard solutions and extracted according to the same procedure applied for the biological samples.…”

Section: Measurement Of Eicosanoid Production By Lc-ms/msmentioning

confidence: 99%

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Unexpected effects of long-term treatment with acetylsalicylic acid on late phase of pulmonary metastasis in murine model of orthotopic breast cancer

et al. 2020

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Long-term administration of acetylsalicylic acid (ASA) was effective in prevention of colorectal cancer, whereas the efficacy of this compound in other cancer types, including breast cancer, has been less convincingly documented. Indeed, the antimetastatic effect of lowdose ASA was observed only in the early intravascular phase of metastasis of breast cancer. In the present work, we characterized the effects of long-term treatment with ASA on the late phase of pulmonary metastasis in a mouse orthotopic 4T1 breast cancer model. Mice were treated with ASA at a dose of 12 mg�kg-1 of body weight daily starting one week prior to inoculation of 4T1 breast cancer cells, and the treatment was continued throughout progression of the disease. ASA administration decreased platelet TXB 2 production in ex vivo assays but did not change thrombin-induced platelet reactivity. Although the number of metastases in the lungs remained unchanged in ASA-treated mice, infiltration of inflammatory cells was increased concomitantly with higher G-CSF and serotonin concentrations in the lungs. Pulmonary NO production was compromised compared to control 4T1 mice. ASA treatment also evoked an increase in platelet and granulocyte counts and decreased systemic NO bioavailability along with increased markers of systemic oxidant stress such as higher GSSG/lower GSH concentrations in RBC. Analysis of eicosanoids in stirred blood demonstrated that administration of ASA at a dose of 12 mg�kg-1 to cancer-bearing mice had an effect beyond inhibition of platelet COX-1, suggesting long-term treatment with lowdose aspirin is not a selective murine platelet COX-1/TXA 2 pathway inhibitor in cancer-bearing mice. In summary, quite surprisingly, long-term treatment with low-dose ASA administered until the advanced phase of breast cancer in a murine orthotopic model of 4T1 breast cancer negatively affected the phenotype of the disease.

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Development and validation of a rapid, specific and sensitive LC-MS/MS bioanalytical method for eicosanoid quantification - assessment of arachidonic acid metabolic pathway activity in hypertensive rats

Cited by 15 publications

References 42 publications

Exogenous Vitamins K Exert Anti-Inflammatory Effects Dissociated from Their Role as Substrates for Synthesis of Endogenous MK-4 in Murine Macrophages Cell Line

Exogenous Vitamins K Exert Anti-Inflammatory Effects Dissociated from Their Role as Substrates for Synthesis of Endogenous MK-4 in Murine Macrophages Cell Line

Thrombin Inhibition Prevents Endothelial Dysfunction and Reverses 20-HETE Overproduction without Affecting Blood Pressure in Angiotensin II-Induced Hypertension in Mice

Unexpected effects of long-term treatment with acetylsalicylic acid on late phase of pulmonary metastasis in murine model of orthotopic breast cancer

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