Effects of aristolochic acid on phospholipase A2 activity and arachidonate metabolism of human neutrophils

Rosenthal, Miriam D.; Vishwanath, Bannikuppe S.; Franson, Richard C.

doi:10.1016/0005-2760(89)90299-3

Cited by 101 publications

(41 citation statements)

References 42 publications

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“…4). Following pre-incubation with the generic cellular PLA 2 inhibitor AA (Vishwanath et al 1988;Rosenthal et al 1989;Chandra et al 2002), no reduction in the LysoPLs production by Tetx was detected, whereas a small decrease (significative with p < 0.05) was observed in the case of Ntx. This latter observation might be because of the slight inhibition of the Ntx enzymatic activity itself by AA as detected by performing an in vitro PLA 2 assay (not shown).…”

Section: Limited Span Hydrolysis Of a Neuroblastoma Cell Linementioning

confidence: 85%

Mass spectrometry analysis of the phospholipase A₂ activity of snake pre‐synaptic neurotoxins in cultured neurons

Paoli

Rigoni

Koster

et al. 2009

Journal of Neurochemistry

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Snake pre-synaptic phospholipase A 2 neurotoxins paralyse the neuromuscular junction by releasing phospholipid hydrolysis products that alter curvature and permeability of the presynaptic membrane. Here, we report results deriving from the first chemical analysis of the action of these neurotoxic phospholipases in neurons, made possible by the use of high sensitivity mass spectrometry. The time-course of the phospholipase A 2 activity (PLA 2 ) hydrolysis of notexin, b-bungarotoxin, taipoxin and textilotoxin acting in cultured neurons was determined. At variance from their enzymatic activities in vitro, these neurotoxins display comparable kinetics of lysophospholipid release in neurons, reconciling the large discrepancy between their in vivo toxicities and their in vitro enzymatic activities. The ratios of the lyso derivatives of phosphatidyl choline, ethanolamine and serine obtained here together with the known distribution of these phospholipids among cell membranes, suggest that most PLA 2 hydrolysis takes place on the cell surface. Although these toxins were recently shown to enter neurons, their intracellular hydrolytic action and the activation of intracellular PLA 2 s appear to contribute little, if any, to the phospholipid hydrolysis measured here. Keywords: lysophospholipids, phospholipase A 2 activity, snake neurotoxins, toxicity. It is still impossible to analyse quantitatively the lipid products released by SPANs at the NMJ, their principal target in humans, for obvious technical reasons. However, it is well established that SPANs are highly toxic when injected into the CNS (Gandolfo et al. 1996;Kolko et al. 1999) and act on isolated brain-derived preparations (Rehm and Betz 1982;Nicholls et al. 1985;Rugolo et al. 1986). Therefore, data obtained with cultured CNS neurons were relevant and were as close to the in vivo situation as is currently experimentally possible. Methods are available to maintain many CNS neurons in primary cultures, but these cultures are mixtures of neurons and glial cells except for the granular neurons of the cerebellum (CGNs, cerebellar granule neurons), wherein cultures consist almost entirely of neurons (Lasher and Zaigon 1972;Levi et al. 1984). We have shown previously that SPANs are very active on these neurons (Rigoni et al. 2004). Using a pure neuronal culture is essential to achieve consistent and reliable MS analysis of changes in lipid compositions, as the presence of a large component of SPAN-resistant glial cells would dilute the changes induced by the toxins. Moreover, glial cells and neurons will have distinct and different compositions of membrane lipid, which will further complicate the MS analysis. CGN neurons are highly sensitive to SPANs and develop a well-defined bulging at axon and dendrite terminals within few minutes from toxin addition; such morphological alteration is accompanied by cytosolic calcium increase at nerve terminals and glutamate release from neurons (Rigoni et al. 2004(Rigoni et al. , 2007. These effects are mimicked by the additi...

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Section: Limited Span Hydrolysis Of a Neuroblastoma Cell Linementioning

confidence: 85%

Mass spectrometry analysis of the phospholipase A₂ activity of snake pre‐synaptic neurotoxins in cultured neurons

Paoli

Rigoni

Koster

et al. 2009

Journal of Neurochemistry

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“…We have therefore tested whether lysoPC, a metabolite generated by PLA2, affects the ability of rab3AL to enhance exocytosis. To concentrate on events downstream of PLA2, spermatozoa were treated with the PLA2 inhibitor aristolochic acid [29,30], a reagent found to inhibit acrosomal exocytosis [13]; test reagents were then included to examine whether they would restore exocytosis.…”

Section: Resultsmentioning

confidence: 99%

rab3‐Peptide stimulates exocytosis of the ram sperm acrosome via interaction with cyclic AMP and phospholipase A₂ metabolites

Garde

Roldán

1996

FEBS Letters

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Aerosomal exocytosis triggered with A23187/Ca 2+ was enhanced by rab3AL, a synthetic peptide corresponding to the effector domain of the small GTP-binding protein rab3. Exocytosis was further enhanced when spermatozoa were also exposed to dihutyryl-cAMP, but was prevented when H-89, a protein kinase A (PKA) inhibitor, was included. The action of rab3AL was not on, or upstream of, phospholipase A2 (PLA2). Inhibition of exocytosis by the PLA2 inhibitor aristolochic acid was overcome by rab3AL when it was included together with lysophosphatidylcholine; this effect was prevented by H-89. These results suggest a functional coupling between rab3 protein, metabolites generated by PLA2, and cAMP-activated PKA, in the final steps leading to membrane fusion during acrosomal exocytosis.Key words: Exocytosis; rab3; Phospholipase Az; Lysophosphatidylcholine; cAMP; Spermatozoa that regulate docking and fusion of secretory granules with the plasma membrane [16][17][18][19].Synthetic peptides corresponding to a putative effector domain of rab3 proteins have been shown to stimulate exocytosis in permeabilized [20 23] and intact cells [24], and membrane fusion in a cell-free assay system [25]. We have therefore used a sperm model system, where cells are stimulated to undergo exocytosis with A23187/Ca 2+, to explore whether rab3 might be involved in the final steps of membrane fusion during acrosomal exocytosis. We present, for the first time, evidence suggesting that rab3 protein, together with PLA2-derived metabolites and the cAMP/PKA signalling pathway, play an important role during the final steps of acrosomal exocytosis in mammalian spermatozoa. Materials and methods

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“…Furthermore, AA was also reported to inhibit Group I PLA 2 in humans with sepsis [11] . From in vitro studies, AA has been shown to suppress phospholipohydration of PLA 2 derived from human synovial fluid, cobra venom, porcine pancreas, and human platelets [12] . The anti-inflammatory activities of AA in different models of inflammation have promoted its use in many countries in herbal formulations for arthritis, rheumatism, gout and chronic inflammatory skin diseases [13,14] .…”

Section: Introductionmentioning

confidence: 99%

Microarray analysis reveals the inhibition of nuclear factor-kappa B signaling by aristolochic acid in normal human kidney (HK-2) cells

Chen

Chiang

et al. 2010

Acta Pharmacol Sin

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Aim: To study the molecular mechanism underlying the effect of aristolochic acid (AA), a major active component of plants from the Aristolochiaceae family using microarray analysis. Methods: Human kidney (HK-2) cells were treated with AA (0, 10, 30, and 90 μmol/L) for 24 h, and the cell viability was measured by a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide assay. Complementary DNA microarrays were used to investigate the gene expression pattern of HK-2 cells exposed to AA in triplicate. A quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assay was used to verify the microarray data for selected nuclear factor kappa B (NF-κB)-regulated genes. Furthermore, the subcellular localization of NF-κB p65 was visualized by immunofluorescence confocal microscopy in HK-2 cells. The NF-κB activity was examined by a luciferase reporter assay in HK-2/NF-κB transgenic cells. Results: AA exhibited a dose-dependent cytotoxic effect in HK-2 cells and induced alterations in the gene expression profiles related to the DNA damage response, DNA repair, macromolecule metabolic process, carbohydrate metabolic process, DNA metabolic process, apoptosis, cell cycle, and transcription. In addition, 9 biological pathways associated with immunomodulatory functions were downregulated in AA-treated HK-2 cells. A network analysis revealed that NF-κB played a central role in the network topology. Among NF-κB-regulated genes, 8 differentially expressed genes were verified by qRT-PCR. The inhibition of NF-κB activity by AA was further confirmed by immunofluorescence confocal microscopy and by NF-κB luciferase reporter assay. Conclusion: Our data revealed that AA could suppress NF-κB activity in normal human cells, perhaps partially accounting for the reported anti-inflammatory effects of some plants from the genus Aristolochia.Keywords: aristolochic acid; microarray analysis; nuclear factor-kappa B; human kidney HK-2 cells; confocal microscopy; luciferase reporter assay Acta Pharmacologica Sinica (2010) 31: 227-236; doi: 10.1038/aps.2009 Original Article # These authors contributed equally to this work. * To whom correspondence should be addressed. [7,8] . AA was found to possess anti-inflammation effects as demonstrated by its ability to inhibit phospholipase A 2 (PLA 2 ) when administered by intramuscular or intraperitoneal injection [9,10] . Furthermore, AA was also reported to inhibit Group I PLA 2 in humans with sepsis [11] . From in vitro studies, AA has been shown to suppress phospholipohydration of PLA 2 derived from human synovial fluid, cobra venom, porcine pancreas, and human platelets [12] . The anti-inflammatory activities of AA in different models of inflammation have promoted its use in many countries in herbal formulations for arthritis, rheumatism, gout and chronic inflammatory skin diseases [13,14] . Moreover, double-blind studies in healthy volunteers show that AA increased the phagocytic activity of peripheral granulocytes after treatment with AA 0.9 mg/d for three to ten consecutive days [...

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Effects of aristolochic acid on phospholipase A2 activity and arachidonate metabolism of human neutrophils

Cited by 101 publications

References 42 publications

Mass spectrometry analysis of the phospholipase A₂ activity of snake pre‐synaptic neurotoxins in cultured neurons

Mass spectrometry analysis of the phospholipase A₂ activity of snake pre‐synaptic neurotoxins in cultured neurons

rab3‐Peptide stimulates exocytosis of the ram sperm acrosome via interaction with cyclic AMP and phospholipase A₂ metabolites

Microarray analysis reveals the inhibition of nuclear factor-kappa B signaling by aristolochic acid in normal human kidney (HK-2) cells

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Effects of aristolochic acid on phospholipase A2 activity and arachidonate metabolism of human neutrophils

Cited by 101 publications

References 42 publications

Mass spectrometry analysis of the phospholipase A2 activity of snake pre‐synaptic neurotoxins in cultured neurons

Mass spectrometry analysis of the phospholipase A2 activity of snake pre‐synaptic neurotoxins in cultured neurons

rab3‐Peptide stimulates exocytosis of the ram sperm acrosome via interaction with cyclic AMP and phospholipase A2 metabolites

Microarray analysis reveals the inhibition of nuclear factor-kappa B signaling by aristolochic acid in normal human kidney (HK-2) cells

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Mass spectrometry analysis of the phospholipase A₂ activity of snake pre‐synaptic neurotoxins in cultured neurons

Mass spectrometry analysis of the phospholipase A₂ activity of snake pre‐synaptic neurotoxins in cultured neurons

rab3‐Peptide stimulates exocytosis of the ram sperm acrosome via interaction with cyclic AMP and phospholipase A₂ metabolites