Peroxiredoxin 6 (Prdx6) is a multifunctional enzyme, a unique member of the peroxiredoxin family, with an important role in antioxidant defense. Moreover, it has also been linked with the biosynthesis of anti-inflammatory and anti-diabetic lipids called fatty acid esters of hydroxy fatty acids (FAHFAs) and many diseases, including cancer, inflammation, and metabolic disorders. Here, we performed metabolomic and lipidomic profiling of subcutaneous adipose tissue from mouse models with genetically modified Prdx6. Deletion of Prdx6 resulted in reduced levels of FAHFAs containing 13-hydroxylinoleic acid (13-HLA). Mutation of Prdx6 C47S impaired the glutathione peroxidase activity and reduced FAHFA levels, while D140A mutation, responsible for phospholipase A2 activity, showed only minor effects. Targeted analysis of oxidized phospholipids and triacylglycerols in adipocytes highlighted a correlation between FAHFA and hydroxy fatty acid production by Prdx6 or glutathione peroxidase 4. FAHFA regioisomer abundance was negatively affected by the Prdx6 deletion, and this effect was more pronounced in longer and more unsaturated FAHFAs. The predicted protein model of Prdx6 suggested that the monomer-dimer transition mechanism might be involved in the repair of longer-chain peroxidized phospholipids bound over two monomers and that the role of Prdx6 in FAHFA synthesis might be restricted to branching positions further from carbon 9. In conclusion, our work linked the peroxidase activity of Prdx6 with the levels of FAHFAs in adipose tissue.
- MeSH
- antioxidancia MeSH
- fosfolipidy MeSH
- mastné kyseliny MeSH
- metabolomika * MeSH
- myši MeSH
- peroxiredoxin VI * genetika MeSH
- peroxiredoxiny MeSH
- tukové buňky MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
To date, the effects of specific modification types and sites on protein lifetime have not been systematically illustrated. Here, we describe a proteomic method, DeltaSILAC, to quantitatively assess the impact of site-specific phosphorylation on the turnover of thousands of proteins in live cells. Based on the accurate and reproducible mass spectrometry-based method, a pulse labeling approach using stable isotope-labeled amino acids in cells (pSILAC), phosphoproteomics, and a unique peptide-level matching strategy, our DeltaSILAC profiling revealed a global, unexpected delaying effect of many phosphosites on protein turnover. We further found that phosphorylated sites accelerating protein turnover are functionally selected for cell fitness, enriched in Cyclin-dependent kinase substrates, and evolutionarily conserved, whereas the glutamic acids surrounding phosphosites significantly delay protein turnover. Our method represents a generalizable approach and provides a rich resource for prioritizing the effects of phosphorylation sites on protein lifetime in the context of cell signaling and disease biology.
- MeSH
- buněčný cyklus fyziologie MeSH
- cyklin-dependentní kinasy genetika metabolismus MeSH
- fosfoproteiny chemie metabolismus MeSH
- fosforylace MeSH
- glutamáty metabolismus MeSH
- hmotnostní spektrometrie metody MeSH
- izotopové značení metody MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- peptidy metabolismus MeSH
- peroxiredoxin VI chemie metabolismus MeSH
- proteolýza * MeSH
- proteom genetika metabolismus MeSH
- proteomika metody MeSH
- sekvence aminokyselin MeSH
- sestřihové faktory chemie metabolismus MeSH
- signální transdukce genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
The patatin-like phospholipase domain containing 3 (PNPLA3) gene (viz. its I148M variant) is one of the key players in the pathogenesis of nonalcoholic fatty liver disease (NAFLD). We have identified a novel insertion/deletion variant of 1114 bp, localized in the second intron of the PNPLA3 gene, which corresponds to the 3' terminal sequence of the long-interspersed element (LINE-1). DNA analysis of 122 NAFLD patients and 167 control subjects as well as RNA analysis of 19 liver biopsies revealed that the novel variant is very common (frequency = 0.41), fully linked to the clinically important I148M variant, and clinically silent. Although the LINE-1 insertion does not seem to have any biological effect, it can impede genotyping of the I148M variant. If insertion prevents the attachment of the diagnostic primer, then the non-insertion allele will be selectively amplified; and thus the frequency of the 148M "risk" allele will be significantly overestimated due to the complete linkage of the LINE-1 insertion and the 148I allele of the PNPLA3 gene. Therefore, our findings underline the importance of careful design and consistent documentation of the methodology, including primer sequences. Critical revisions of the results of some studies that have already been reported may therefore be needed.
- MeSH
- acyltransferasy genetika MeSH
- alely MeSH
- dlouhé rozptýlené jaderné elementy genetika MeSH
- fosfolipasy A2 nezávislé na vápníku genetika MeSH
- genetická predispozice k nemoci genetika MeSH
- genotyp MeSH
- játra patologie MeSH
- jednonukleotidový polymorfismus genetika MeSH
- lidé MeSH
- nealkoholová steatóza jater genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The human iPSC cell lines, PLANFiPS1-Sv4F-1 (RCPFi004-A), PLANFiPS2-Sv4F-1 (RCPFi005-A), PLANFiPS3-Sv4F-1 RCPFi006-A), derived from dermal fibroblast from three patients suffering PLAN (PLA2G6-associated neurodegeneration; MIM 256600) caused by mutations in the PLA2G6 gene, was generated by non-integrative reprogramming technology using OCT3/4, SOX2, CMYC and KLF4 reprogramming factors. The pluripotency was assessed by immunocytochemistry and RT-PCR. Differentiation capacity was verified in vitro. This iPSC line can be further differentiated toward affected cells to better understand molecular mechanisms of disease and pathophysiology.
The relationship between baroreflex sensitivity (BRS) and inflammatory vascular biomarker Lipoprotein associated phospholipase A2 (Lp-PLA2) in subjects with high normal blood pressure (HNBP, prehypertensives) with a positive family history of hypertension (FHH+) and hypertension history free control subjects (FHH-) was evaluated. A total of 24 HNBP participants (age 39.5 ± 2.5 years, 18 male/ 6 female) were studied. 14 HNBP subjects FHH+ were compared to 10 HNBP participants FHH-, being of similar age and body mass index. BRS (ms/mmHg) was determined by the sequence and spectral methods (five-minute non-invasive beat-to-beat recording of blood pressure and RR interval, controlled breathing at a frequency of 0.33 Hz). Venous blood was analyzed for Lp-PLA2 biomarker of vascular inflammation and atherothrombotic activity. A significant negative correlation between spontaneous BRS obtained by both methods and systolic blood pressure (BP) was present (BRS spect r = -0.54, P<0.001, BRS seq r = -0.59, P<0.001). BRS obtained by sequence and spectral methods were reduced in HNBP FHH+ compared to the group of HNBP FHH- (P = 0.0317 BRS seq, P = 0.0395 BRS spect). Lp-PLA2 was significantly higher in HNBP FHH+ compared to FHH- controls (P<0.05). Lp-PLA2 was negatively correlated with BRS obtained by sequence method (r = -0.798, R2 = 0.636, P<0.001) in the HNBP FHH+ subjects. These findings demonstrate that reduced baroreflex sensitivity, as a marker of autonomic dysfunction, is associated with vascular inflammation, predominantly in otherwise healthy participants with a positive family history of hypertension who could predispose to increased risk of hypertension. We conclude that our transversal study suggests that a lowbaroreflex sensitivity could be an early sign of autonomic dysfunction even in the prehypertensive period, and to corroborate these findings, a longitudinal study is needed.
- MeSH
- 1-alkyl-2-acetylglycerofosfocholinesterasa krev MeSH
- anamnéza MeSH
- autonomní nervový systém patofyziologie MeSH
- baroreflex * MeSH
- biologické markery krev MeSH
- dospělí MeSH
- hypertenze krev diagnóza enzymologie patofyziologie MeSH
- krevní tlak * MeSH
- lidé středního věku MeSH
- lidé MeSH
- prediktivní hodnota testů MeSH
- rizikové faktory MeSH
- studie případů a kontrol MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Obesity is believed to be associated with a dysregulated endocannabinoid system which may reflect enhanced inflammation. However, reports of this in human white adipose tissue (WAT) are limited and inconclusive. Marine long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFAs) have anti-inflammatory actions and therefore may improve obesity-associated adipose tissue inflammation. Therefore, fatty acid (FA) concentrations, endocannabinoid concentrations, and gene expression were assessed in subcutaneous WAT (scWAT) biopsies from healthy normal weight individuals (BMI 18.5-25 kg/m2) and individuals living with metabolically healthy obesity (BMI 30-40 kg/m2) prior to and following a 12-week intervention with 3 g fish oil/day (1.1 g eicosapentaenoic acid (EPA) + 0.8 g DHA) or 3 g corn oil/day (placebo). WAT from individuals living with metabolically healthy obesity had higher n-6 PUFAs and EPA, higher concentrations of two endocannabinoids (anandamide (AEA) and eicosapentaenoyl ethanolamide (EPEA)), higher expression of phospholipase A2 Group IID (PLA2G2D) and phospholipase A2 Group IVA (PLA2G4A), and lower expression of CNR1. In response to fish oil intervention, WAT EPA increased to a similar extent in both BMI groups, and WAT DHA increased by a greater extent in normal weight individuals. WAT EPEA and docosahexaenoyl ethanolamide (DHEA) increased in normal weight individuals only and WAT 2-arachidonyl glycerol (2-AG) decreased in individuals living with metabolically healthy obesity only. Altered WAT fatty acid, endocannabinoid, and gene expression profiles in metabolically healthy obesity at baseline may be linked. WAT incorporates n-3 PUFAs when their intake is increased which affects the endocannabinoid system; however, effects appear greater in normal weight individuals than in those living with metabolically healthy obesity.
- MeSH
- časové faktory MeSH
- dospělí MeSH
- dvojitá slepá metoda MeSH
- endokanabinoidy metabolismus MeSH
- fixní kombinace léků MeSH
- fosfolipasy A2, skupina II metabolismus MeSH
- fosfolipasy A2, skupina IV metabolismus MeSH
- kyselina eikosapentaenová aplikace a dávkování MeSH
- kyseliny arachidonové metabolismus MeSH
- kyseliny dokosahexaenové aplikace a dávkování MeSH
- lidé středního věku MeSH
- lidé MeSH
- metabolicky zdravá obezita diagnóza farmakoterapie metabolismus MeSH
- mladiství MeSH
- mladý dospělý MeSH
- podkožní tuk účinky léků metabolismus MeSH
- polynenasycené alkamidy metabolismus MeSH
- potravní doplňky * MeSH
- receptor kanabinoidní CB1 metabolismus MeSH
- výsledek terapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- randomizované kontrolované studie MeSH
- Geografické názvy
- Anglie MeSH
Lipoprotein-asociovaná fosfolipáza A2 (Lp-PLA2) je silně exprimována v pokročilých stadiích aterosklerózy a může hrát roli v nestabilitě aterosklerotického pásu. Vybrali jsme skupinu seniorů, kteří podstoupili transkatétrovou implantaci aortální chlopně (TAVI) nebo balónkovou angioplastiku (BA) a rozdělili je do dvou skupin (diabetici a nediabetici). Následně jsme mezi nimi porovnali koncentraci Lp-PLA2. Metodika: Do studie bylo zahrnuto 44 pacientů ve věku 79,6 ± 5,6 roku s těžkou významnou aortální stenózou. 35 z nich podstoupilo TAVI, zbytek BA. U 21 pacientů byl potvrzen diabetes mellitus. V kontrolní skupině zdravých seniorů (pacienti bez významnějších komorbidit) bylo 48 subjektů. Koncentrace Lp-PLA2 se měřila pomocí ELISA metody, a to před zákrokem a 3 dny po něm. Výsledky: Hladina Lp-PLA2 byla u pacientů s aortální stenózou výrazně elevována (1296 ± 437 ng/ml před TAVI, 1413 ± 268 ng/ml před BA) a po výkonu se ještě zvýšila; po TAVI 1604 ± 437 ng/ml (P < 0,01) a po BA 1808 ± 303 ng/ml (P < 0,01). Zároveň ve skupině diabetiků byla hladina Lp-PLA2 signifikantně vyšší než u nemocných bez diabetu. Závěr: U starších pacientů by se Lp-PLA2 mohla stát novým biomarkerem přítomnosti aterosklerotické léze hrozící rupturou plátu.
Lipoprotein-associated phospholipase A2 (Lp-PLA2) is extensively expressed by advanced atherosclerotic lesions and may play a role in plaque instability. We selected a group of elderly subjects that underwent transcatheter aortic valve implantation (TAVI) or balloon angioplasty (BA) and separated them into two groups, diabetic and nondiabetic, to compare the level of Lp-PLA2 mass between them. Methods: 44 patients aged 79.6 ± 5.6 years with symptomatic severe aortic valve stenosis underwent TAVI (n = 35) or BA (n = 9). 21 subjects had confirmed type 2 diabetes mellitus. Lp-PLA2 mass was measured using an enzyme- linked immunosorbent assay kit (USCN Life Science, China) before and 3 days after the procedure. Results: Lp-PLA2 mass was significantly elevated in this population (1296 ± 358 ng/mL before TAVI; 1413 ± 268 ng/mL before BA) and further increased after TAVI (1604 ± 437 ng/mL, P < 0.01) or BA (1808 ± 303 ng/mL, P < 0.01). Lp-PLA2 mass was significantly increased on the diabetic group before these interventions. Conclusion: Lp-PLA2 may be a novel biomarker for the presence of rupture-prone atherosclerotic lesions in elderly patients. Levels of Lp-PLA2 in diabetic patients may accompany the higher amount of small dense LDL particles seen in these subjects.
- Klíčová slova
- lipoprotein-asociovaná fosfolipáza A2, transkatetrová implantace.,
- MeSH
- aortální chlopeň patologie MeSH
- aortální stenóza etiologie MeSH
- ateroskleróza * etiologie MeSH
- balónková angioplastika MeSH
- diabetes mellitus MeSH
- fosfolipasy A2 * analýza krev MeSH
- klinická studie jako téma MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
The inflammatory process is a natural self-defense response of the organism to damage agents and its action mechanism involves a series of complex reactions. However, in some cases, this process can become chronic, causing much harm to the body. Therefore, over the years, many anti-inflammatory drugs have been developed aiming to decrease the concentrations of inflammatory mediators in the organism, which is a way of controlling these abnormal chain reactions. The main target of conventional anti-inflammatory drugs is the cyclooxygenase (COX) enzyme, but its use implies several side effects. Thus, based on these limitations, many studies have been performed, aiming to create new drugs, with new action mechanisms. In this sense, the phospholipase A2 (PLA2) enzymes stand out. Among all the existing isoforms, secretory PLA2 is the major target for inhibitor development, since many studies have proven that this enzyme participates in various inflammatory conditions, such as cancer, Alzheimer and arthritis. Finally, for the purpose of developing anti-inflammatory drugs that are sPLA2 inhibitors, many molecules have been designed. Accordingly, this work presents an overview of inflammatory processes and mediators, the current available anti-inflammatory drugs, and it briefly covers the PLA2 enzymes, as well as the diverse structural array of the newest sPLA2 inhibitors as a possible target for the production of new anti-inflammatory drugs.
- MeSH
- antiflogistika farmakologie MeSH
- cyklooxygenasa 2 MeSH
- fosfolipasy A2 MeSH
- inhibitory enzymů MeSH
- inhibitory fosfolipasy A2 MeSH
- lidé MeSH
- nádory MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Kardiovaskulární onemocnění jsou ve vyspělých zemích stále vedoucí příčinou morbidity a mortality geriatrických pacientů. Patofyziologickým podkladem těchto chorob je majoritně ateroskleróza. Proto je včasné odhalení potencionálních rizikových faktorů aterosklerózy považováno za stěžejní bod v prevenci a časné diagnóze kardiovaskulárních onemocnění. V posledních letech bylo objeveno několik nových markerů, nicméně jejich přínos v kontextu s klinickou praxí (stratifikace rizika, terapeutické intervence aj.) zůstává stále sporný. Tento text shrnuje vybrané novější vyšetřovací metody a markery aterosklerózy u geriatrických pacientů se zaměřením na lipoprotein-asociovanou fosfolipázu A2, kde autoři prezentují i vlastní zkušenosti.
The leading cause of death in developed countries are still complications of cardiovascular diseases. Atherosclerosis is one of the main risk factors and the main pathophysiological mechanism of these diseases. It is thus of paramount importance to identify potential risk factors of atherosclerosis at early stages, or even as a part of primary prevention of cardiovascular diseases. In the past years many new markers were explored, however their value in the context of clinical praxis (risk stratification, therapeutic interventions…) is still questionable. This paper tries to elucidate on the available methods to examine atherosclerosis, focusing especially on the geriatric population, where this issue is very common.
- MeSH
- arterioskleróza * diagnóza MeSH
- biologické markery MeSH
- fosfolipasy A2 * analýza MeSH
- kardiovaskulární nemoci diagnóza MeSH
- křehký senior MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Ciel: Fosfolipáza A2 viazaná na lipoproteíny (Lp-PLA2) je enzým kumulujúci sa v aterosklerotickom pláte, ktorý vyvoláva zápal plátu, čo môže viesť k ruptúre plátu. Cieľom práce je porovnať hodnoty sérovej koncentrácie Lp-PLA2 medzi skupinou zdravých jedincov a skupinami ľudí s arteriálnou hypertenziou (AH), ischemickou chorobou srdca (ICHS) a ischemickou cievnou mozgovou príhodou (iCMP) a tým posúdiť vhodnosť Lp-PLA2 ako biomarkera rizika iCMP. AH a ICHS srdca sú považované za rizikové faktory iCMP, preto sme sérovú koncentráciu Lp-PLA2 merali aj u pacientov s týmito ochoreniami. Metódy: Sérovú koncentráciu Lp-PLA2 sme vyšetrovali enzýmovou imunoanalýzou metódou sedvičovej ELISA diaDexus PLAC® Test ELISA Kit (Diadexus, Inc., San Francisco, USA). Štatistické spracovanie súboru sme previedli v programe IBM SPSS Statistics 24 (IBM Corp., New York, USA) použitím Fisherovho exaktného testu a neparametrických korelácií. Výsledky: Celkovo sme vyšetrili 401 jedincov (muži 43 %), v skupine zdravých ľudí 80 jedincov (muži 35 %), v skupine s AH 96 jedincov (muži 43 %), v skupine s ICHS 85 jedincov (muži 39 %) a v skupine s iCMP 140 jedincov (muži 49 %). Sérová koncentrácia Lp-PLA2 v skupine zdravých ľudí bola signifikantne nižšia ako v skupine s AH (p = 0 × 10–3), ICHS (p = 0 × 10–3) a iCMP (p = 0 × 10–3). Záver: Naša štúdia potvrdila predpoklad, že ľudia s AH, ICHS a iCMP majú vyššiu sérovú koncentráciu Lp-PLA2 ako zdraví jedinci, a teda zvýšenú pravdepodobnosť výskytu zapálených aterosklerotických plátov s rizikom ruptúry ako zdraví ľudia. Ale zvýšená hladina Lp-PLA2 sa u ľudí s AH, ICHS a iCMP v našom súbore vyskytovala aj napriek statínovej liečbe, čo nás viedlo k záveru, že úloha Lp-PLA2 pri vzniku a prehlbovaní zápalu aterosklerotického plátu môže byť zložitejšia ako len hydrolýza oxidovaných LDL v aterosklerotickom pláte.
Aim: Lipoprotein-associated phospholipase A2 (Lp-PLA2) is an enzyme accumulated in atherosclerotic plaques and causes plaques inflammation that can induce plaque rupture. The aim of this work is to compare serum Lp-PLA2 concentration in healthy subjects and in patients with arterial hypertension (AH), ischemic heart disease (IHD) and ischemic stroke (IS) so that we could assess utility of Lp-PLA2 as a biomarker for IS risk. AH and IHD are considered risk factors for IS, therefore we measured serum Lp-PLA2 concentration also in patients with these diseases. Methods: Serum Lp-PLA2 concentration was determined by diaDexus PLAC® Test ELISA Kit (Diadexus, Inc., San Francisco, USA), a sandwich enzyme immunoassay. The statistical analysis was performed with IBM SPSS Statistics 24 (IBM Corp., New York, USA) using the Fisher’s exact test and non-parametric correlations. Results: Our cohort comprised of 401 subjects in total (43% males), 80 subjects in the group of healthy individuals (35% males), 96 subjects in the group with AH (43% males), 85 subjects in the group with IHD (39% males) and 140 subjects in the group with IS (49% males). Serum Lp-PLA2 concentration in the group of healthy individuals was significantly lower than that in the group with AH (p = 0 × 10–3), IHD (p = 0 × 10–3) and IS (p = 0 × 10–3). Conclusion: Our study confirmed the assumption that people with AH, IHD and IS have higher levels of serum Lp-PLA2 concentration than healthy people hence a higher incidence of inflamed atherosclerotic plaques and higher risk of rupture of these plaques, but a higher level of serum Lp-PLA2 persisted in people with AH, IHD and IS in our cohort despite the statin therapy, leading us to conclude that the role of Lp-PLA2 in the development and intensification of atherosclerotic plaque inflammation may be more complicated than only the hydrolysis of oxidized LDL in atherosclerotic plaque.
