scholarly journals Comparison of extracellular ATP and UTP signalling in rat renal mesangial cells. No indications for the involvement of separate purino- and pyrimidino-ceptors

1990 ◽  
Vol 272 (2) ◽  
pp. 469-472 ◽  
Author(s):  
J Pfeilschifter
Keyword(s):  
Pertussis Toxin ◽  
Mesangial Cells ◽  
Partial Agonist ◽  
Peptide Hormone ◽  
Extracellular Atp ◽  
Phosphoinositide Hydrolysis ◽  
Response Curves ◽  
Reactive Blue 2 ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition

Extracellular ATP and UTP caused a rapid formation of InsP3, with similar kinetics and dose-dependences. ITP also displayed strong agonistic properties in terms of InsP3 production, whereas CTP was almost inactive. Pretreatment of the cells with pertussis toxin attenuated ATP- and UTP-stimulated InsP3 generation to a comparable extent, indicating that both nucleotides couple to phospholipase C by a pertussis-toxin-sensitive G-protein. Short-term (15 min) treatment of the cells with phorbol 12-myristate 13-acetate (PMA) produced a dose-dependent inhibition of ATP- and UTP-induced InsP3 formation. Furthermore, down-regulation of protein kinase C by long-term (24 h) exposure of the cells to PMA resulted in a comparable potentiation of phosphoinositide hydrolysis by both nucleotides. Preincubation of mesangial cells with ATP or UTP caused a pronounced cross-desensitization of subsequent nucleotide-stimulated InsP3 production. ATP and UTP displayed no additivity in terms of InsP3 formation, when used at maximally effective concentrations. In contrast, the peptide hormone angiotensin II interacted in an additive manner with either nucleotide in stimulating phosphoinositide hydrolysis. Reactive Blue 2, a putative P2y-purinoceptor antagonist, caused a rightward shift of both the ATP and UTP dose-response curves. However, since 2-methylthio-ATP was only a partial agonist in stimulating InsP3 formation, the mesangial-cell ATP receptor appears to be different from a classic P2y-receptor. In summary, these results provide no evidence for separate purino- and pyrimidino-ceptors on mesangial cells. In contrast, ATP and UTP may use a common nucleotide receptor for transducing their signals in mesangial cells.

Haloperidol-mediated phosphoinositide hydrolysis via direct activation of α1-adrenoceptors in frontal cerebral rat cortex

1999 ◽  
Vol 77 (1) ◽  
pp. 22-28 ◽  
Author(s):  
Tania Gabriela Borda ◽  
Graciela Cremaschi ◽  
Leonor Sterin-Borda
Keyword(s):  
Inositol Phosphate ◽  
Partial Agonist ◽  
Phosphoinositide Hydrolysis ◽  
Maximal Effect ◽  
Response Curves ◽  
Pi Turnover ◽  
Direct Activation ◽  
Adrenoceptor Blocker ◽  
6 Hydroxydopamine ◽  
Pi Hydrolysis

In addition to its effect on D2 dopamine receptor blockades, haloperidol is able to interact with multiple neurotransmitters (NTs). Its action on phosphoinositide (PI) turnover was studied on cerebral cortex preparations. It induces an increase in inositol phosphate (IP) accumulation, which was only blunted by the α1-adrenoceptor blocker prazosin. Haloperidol maximal effect (Emax) was less than the effect of the full agonist norepinephrine (NE), and dose-response curves for both NE in the presence of submaximal doses of haloperidol and haloperidol in the presence of Emax doses of NE showed that haloperidol behaves as a partial agonist of cerebral α1-adrenoceptors. Its effect on PI hydrolysis is mediated through phospholipase C activation, as 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate (NCDC) and 1-[6-([(17β)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino)hexyl]-1H-pyrrole-2,5-dione) (U-73122) were able to abrogate both haloperidol and NE actions. Further, the typical neuroleptic exerts a direct activation of α1-adrenoceptors as its actions were not modified by cocaine and persisted in spite of chemical rat cerebral denervation with 6-hydroxydopamine (6-OHDA). The possibility that this agonistic action on α1-adrenoceptors would be involved in haloperidol side effects is also discussed.Key words: haloperidol, neuroleptics, α1-adrenoceptor, phosphoinositide hydrolysis, cerebral frontal cortex.

scholarly journals Acute inhibition of the betaine transporter by ATP and adenosine in renal MDCK cells

2008 ◽  
Vol 295 (1) ◽  
pp. F108-F117 ◽  
Author(s):  
Stephen A. Kempson ◽  
Jason M. Edwards ◽  
Alyssa Osborn ◽  
Michael Sturek
Keyword(s):  
Atp Hydrolysis ◽  
Mdck Cells ◽  
Extracellular Atp ◽  
Mechanical Stimuli ◽  
Autocrine Signaling ◽  
Dependent Inhibition ◽  
Intracellular Ca ◽  
Inhibitory G Protein ◽  
Dose Dependent Inhibition ◽  
Immunohistochemical Studies

Extracellular ATP interacts with purinergic P2 receptors to regulate a range of physiological responses, including downregulation of transport activity in the nephron. ATP is released from cells by mechanical stimuli such as cell volume changes, and autocrine signaling by extracellular ATP could occur in renal medullary cells during diuresis. This was tested in Madin-Darby canine kidney (MDCK) cells, a model used frequently to study P1 and P2 receptor activity. ATP was released within 1 min after transfer from 500 to 300 mosmol/kgH2O medium. A 30-min incubation with ATP produced dose-dependent inhibition (0.01–0.10 mM) of the renal betaine/GABA transporter (BGT1) with little effect on other osmolyte transporters. Inhibition was reproduced by specific agonists for P2X (α,β-methylene-ATP) and P2Y (UTP) receptors. Adenosine, the final product of ATP hydrolysis, also inhibited BGT1 but not taurine transport. Inhibition by ATP and adenosine was blocked by pertussis toxin and A73122, suggesting involvement of inhibitory G protein and PLC in postreceptor signaling. Both ATP and adenosine (0.1 mM) produced rapid increases in intracellular Ca2+, due to the mobilization of intracellular Ca2+ stores and Ca2+ influx. Blocking these Ca2+ increases with BAPTA-AM also blocked the action of ATP and adenosine on BGT1 transport. Finally, immunohistochemical studies indicated that inhibition of BGT1 transport may be due to endocytic accumulation of BGT1 proteins from the plasma membrane. We conclude that ATP and adenosine, through stimulation of PLC and intracellular Ca2+, may be rapidly acting regulators of BGT1 transport especially in response to a fall in extracellular osmolarity.

Inhibition of rat mesangial cell growth by heparan sulfate

1990 ◽  
Vol 258 (2) ◽  
pp. F259-F265
Author(s):  
G. C. Groggel ◽  
G. N. Marinides ◽  
P. Hovingh ◽  
E. Hammond ◽  
A. Linker
Keyword(s):  
Cell Growth ◽  
Heparan Sulfate ◽  
Mesangial Cell ◽  
Mesangial Cells ◽  
Significant Inhibition ◽  
3T3 Fibroblasts ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Dose Dependent ◽  
Endogenous Component

The ability of heparan sulfate, an endogenous component of the glomerulus, to regulate the growth of cultured rat mesangial cells was investigated. Heparan sulfate caused a dose-dependent inhibition of rat mesangial cell growth, 85% inhibition compared with controls at the highest dose (1,000 micrograms/ml). Chondroitin sulfate produced no inhibition. The low-sulfated fraction of heparan sulfate (9%) produced more inhibition than the high-sulfated fraction (13%), 90 +/- 1 vs. 71 +/- 2% (P = 0.002). The effects of the heparan sulfate were completely reversible. Treatment of heparan sulfate with heparitinase increased the degree of inhibition, 71 +/- 1 vs. 84 +/- 1% (P less than 0.001). Four different oligosaccharides derived from heparan sulfate and heparin were tested for their ability to inhibit growth. One of the oligosaccharides, low-sulfated (10%), caused significant inhibition, 76 +/- 2%. Heparan sulfate was also able to inhibit the growth of Swiss 3T3 fibroblasts (63 +/- 5%). This inhibition was less marked than that seen with mesangial cells. Thus heparan sulfate was able to significantly inhibit rat mesangial cell growth in culture. Alterations in glomerular heparan sulfate may play an important role in alterations in mesangial cell growth.

scholarly journals Different effects of phorbol ester on angiotensin II- and stable GTP analogue-induced activation of polyphosphoinositide phosphodiesterase in membranes isolated from rat renal mesangial cells

1987 ◽  
Vol 248 (1) ◽  
pp. 209-215 ◽  
Author(s):  
J Pfeilschifter ◽  
C Bauer
Keyword(s):  
Protein Kinase C ◽  
Angiotensin Ii ◽  
Protein Kinase ◽  
Phorbol Ester ◽  
Mesangial Cells ◽  
Angiotensin Ii Receptor ◽  
Kinase C ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Gamma S

Pretreatment with pertussis toxin inhibits angiotensin II-induced activation of polyphosphoinositide phosphodiesterase in rat renal mesangial cells [Pfeilschifter & Bauer (1986) Biochem. J. 236, 289-294]. Furthermore, activation of protein kinase C by the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) and by 1-oleoyl-2-acetylglycerol (OAG) abolishes angiotensin II-induced formation of inositol trisphosphate (IP3) in mesangial cells [Pfeilschifter (1986) FEBS Lett. 203, 262-266]. Using membrane preparations of [3H]inositol-labelled mesangial cells we tried to obtain further insight as to the step at which protein kinase C might interfere with the signal transduction mechanism in mesangial cells. Angiotensin II (100 nM) stimulates IP3 formation from membrane preparations of [3H]inositol-labelled mesangial cells with a half-maximal potency of 1.1 nM. The angiotensin II-induced formation of IP3 is enhanced by GTP. This effect of angiotensin II is completely blocked by the competitive antagonist [Sar1,Ala8]angiotensin II. Guanosine 5′-[gamma-thio]triphosphate (GTP gamma S) and guanosine 5′-[beta gamma-imido]triphosphate (Gpp[NH]p), non-hydrolysable analogues of GTP, stimulate IP3 production in the absence of angiotensin II with Kd values of 0.19 microM and 2.4 microM, respectively. Angiotensin II augments the increase in IP3 formation induced by GTP gamma S. However, when mesangial cells were pretreated with TPA there was a dose-dependent inhibition of the synergistic action of angiotensin II on GTP gamma S-induced IP3 production. Comparable results are obtained with OAG, while the non-tumour-promoting phorbol ester 4 alpha-phorbol 12,13-didecanoate is without effect. These results suggest that activation of protein kinase C in mesangial cells does not impair phosphoinositide hydrolysis by stable GTP analogues but somehow seems to interfere with the stimulatory interaction of the occupied angiotensin II receptor with the transducing G-protein.

LiCl and CCK inhibit gastric emptying and feeding and stimulate OT secretion in rats

1989 ◽  
Vol 256 (2) ◽  
pp. R463-R468 ◽  
Author(s):  
M. J. McCann ◽  
J. G. Verbalis ◽  
E. M. Stricker
Keyword(s):  
Gastric Emptying ◽  
Food Intake ◽  
Peptide Hormone ◽  
Autonomic Functions ◽  
Systemic Injection ◽  
Similar Relation ◽  
Dependent Inhibition ◽  
Parvocellular Neurons ◽  
Dose Dependent Inhibition ◽  
Dose Dependent

Systemic injection of the nauseogenic agent LiCl is known to increase neurohypophyseal secretion of oxytocin (OT) in rats. The present results indicated that the induced OT secretion was related exponentially to the inhibition of food intake. A similar relation between OT secretion and food intake also was observed after systemic injection of the peptide hormone cholecystokinin (CCK). However, the effects of each agent on food intake lasted much longer than the observed increases in OT secretion. In contrast, both LiCl and CCK produced a dose-dependent inhibition of gastric emptying in rats that was closely related temporally to the inhibition of food intake. The similarity of these three responses to LiCl and CCK suggests that both agents stimulate common central mechanisms, apparently involving both magnocellular and parvocellular neurons in the paraventricular nucleus of the hypothalamus, whereby these neuroendocrine, behavioral, and autonomic functions are integrated.

Peptides PHI and VIP: comparison between vascular and nonvascular smooth muscle effect in rabbit uterus

1986 ◽  
Vol 251 (1) ◽  
pp. E48-E51 ◽  
Author(s):  
B. Bardrum ◽  
B. Ottesen ◽  
J. Fahrenkrug
Keyword(s):  
Smooth Muscle ◽  
Muscle Activity ◽  
Combined Effect ◽  
Urogenital Tract ◽  
Mechanical Activity ◽  
Response Curves ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Dose Dependent

The distribution and effects of the two neuropeptides, vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine amide (PHI), on vascular and nonvascular smooth muscle in the urogenital tract of nonpregnant rabbit female, were investigated. Immunoreactive VIP and PHI were present in all regions except the ovary with the highest concentration in the uterine cervix. By using in vitro tension recordings of myometrial specimens, it was demonstrated that both peptides displayed a dose-dependent inhibition of the mechanical activity. The dose-response curves of VIP and PHI were superimposable with and ID50 of 3 X 10(-8) mol/l, and their combined effect was additive. In addition, the influence of the two peptides on myometrial blood flow (MBF) was investigated by the xenon-133 washout technique. Both peptides were found to increase MBF with the same potency and efficacy. Their combined effect was additive. In conclusion VIP and PHI are present in the rabbit urogenital tract, and the two peptides are equipotent inhibitors of mechanical nonvascular and vascular smooth muscle activity in the uterus.

Differences between antigenic determinants of pig and cat zona pellucida proteins

Reproduction ◽  
2000 ◽  
pp. 15-23 ◽  
Author(s):  
K Jewgenow ◽  
M Rohleder ◽  
I Wegner
Keyword(s):  
In Vitro Fertilization ◽  
Zona Pellucida ◽  
Antigenic Determinants ◽  
Vitro Fertilization ◽  
Contraceptive Vaccine ◽  
Sperm Binding ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Dose Dependent

Despite many efforts, the control of reproduction in feral cat populations is still a problem in urban regions around the world. Immunocontraception is a promising approach; thus the present study examined the suitability of the widely used pig zona pellucida proteins (pZP) for contraception in feral domestic cats. Purified zona pellucida proteins obtained from pig and cat ovaries were used to produce highly specific antisera in rabbits. Antibodies against pZP raised in rabbits or lions were not effective inhibitors of either in vitro sperm binding (cat spermatozoa to cat oocytes) or in vitro fertilization in cats, whereas antibodies against feline zona pellucida proteins (fZP) raised in rabbits showed a dose-dependent inhibition of in vitro fertilization. Immunoelectrophoresis, ELISA and immunohistology of ovaries confirmed these results, showing crossreactivity of anti-fZP sera to fZP and to a lesser extent to pZP, but no interaction of anti-pZP sera with fZP. It is concluded that cat and pig zonae pellucidae express a very small number of shared antigenic determinants, making the use of pZP vaccine in cats questionable. A contraceptive vaccine based on feline zona pellucida determinants will be a better choice for the control of reproduction in feral cats if immunogenity can be achieved.

Antioxidant effects and in vitro cytotoxicity on human cancer cell lines of flavonoid-rich Flamboyant (Delonix regia (Bojer) Raf.) flower extract

2020 ◽  
Vol 21 ◽  
Author(s):  
Putthiporn Khongkaew ◽  
Phanphen Wattanaarsakit ◽  
Konstantinos I. Papadopoulos ◽  
Watcharaphong Chaemsawang
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Flower Extract ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Murine Leukemia Cell ◽  
Dose Dependent

Background: Cancer is a noncommunicable disease with increasing incidence and mortality rates both worldwide and in Thailand. Its apparent lack of effective treatments is posing challenging public health issues. Introduction: Encouraging research results indicating probable anti-cancer properties of the Delonix regia flower extract (DRE) have prompted us to evaluate the feasibility of developing a type of product for future cancer prevention or treatment. Methods and Results: In the present report, using High Performance Liquid Chromatography (HPLC), we demonstrate in the DRE, the presence of high concentrations of three identifiable flavonoids, namely rutin 4.15±0.30 % w/w, isoquercitrin 3.04±0.02 %w/w, and myricetin 2.61±0.01 % w/w respectively while the IC50 of DPPH and ABTS assay antioxidation activity was 66.88±6.30 µg/ml and 53.65±7.24 µg/ml respectively. Discussion: Our cancer cell line studies using the MTT assay demonstrated DREs potent and dose dependent inhibition of murine leukemia cell line (P-388: 35.28±4.07% of cell viability remaining), as well as of human breast adenocarcinoma (MCF-7), human cervical carcinoma (HeLa), human oral cavity carcinoma (KB), and human colon carcinoma (HT-29) cell lines in that order of magnitude. Conclusion: Three identifiable flavonoids (rutin, isoquercitrin and myricetin) with high antioxidation activity and potent and dose dependent inhibition of murine leukemia cell line and five other cancer cell lines were documented in the DRE. The extract’s lack of cytotoxicity in 3 normal cell lines is a rare advantage not usually seen in current antineoplastic agents. Yet another challenge of the DRE was its low dissolution rate and long-term storage stability, issues to be resolved before a future product can be formulated.

Investigation on the antibacterial activity of electronic cigarette liquids (ECLs): a proof of concept study

2020 ◽  
Vol 21 ◽  
Author(s):  
Virginia Fuochi ◽  
Massimo Caruso ◽  
Rosalia Emma ◽  
Aldo Stivala ◽  
Riccardo Polosa ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Bacterial Species ◽  
A549 Cells ◽  
Bactericidal Effect ◽  
Minimal Bactericidal Concentration ◽  
Viability Assay ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Dose Dependent

Background: The key ingredients of e-cigarettes liquid are commonly propane-1,2-diol (also called propylene glycol) and propane-1,2,3-triol (vegetal glycerol) and their antimicrobial effects are already established. The nicotine and flavors which are often present in e-liquids can interfere with the growth of some microorganisms. Objective: The effect of the combining these elements in e-liquids is unknown. The aim of the study was to investigate the possible effects of these liquids on bacterial growth in the presence or absence of nicotine and flavors. Methods: Susceptibilities of pathogenic strains (Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli, Enterococcus faecalis and Sarcina lutea) were studied by means of a multidisciplinary approach. Cell viability and antioxidant assays were also evaluated. Results: All e-liquids investigated showed antibacterial activity against at least one pathogenic strain. A higher activity was correlated to the presence of flavors and nicotine. Discussion: In most cases the value of minimal bactericidal concentration is equal to the value of minimal inhibitory concentration showing that these substances have a bactericidal effect. This effect was observed in concentrations up to 6.25% v/v. Antioxidant activity was also correlated to presence of flavors. Over time, the viability assay in human epithelial lung A549 cells showed a dose-dependent inhibition of cell growth. Conclusion: Our results have shown that flavors considerably enhance the antibacterial activity of propane-1,2-diol and propane-1,2,3-triol. This study provides important evidence that should be taken into consideration in further investigative approaches, to clarify the different sensitivity of the various bacterial species to e-liquids, including the respiratory microbiota, to highlight the possible role of flavors and nicotine.

scholarly journals In VitroActivities of Hexaazatrinaphthylenes against Leishmania spp.

2015 ◽  
Vol 59 (5) ◽  
pp. 2867-2874 ◽  
Author(s):  
Atteneri López-Arencibia ◽  
Daniel García-Velázquez ◽  
Carmen M. Martín-Navarro ◽  
Ines Sifaoui ◽  
María Reyes-Batlle ◽  
...  
Keyword(s):  
Therapeutic Agents ◽  
Content Type ◽  
Inhibition Effect ◽  
Intracellular Amastigotes ◽  
Dependent Inhibition ◽  
Leishmania Spp ◽  
Dose Dependent Inhibition ◽  
Dose Dependent ◽  
Potential Use

ABSTRACTThein vitroactivity of a novel group of compounds, hexaazatrinaphthylene derivatives, against two species ofLeishmaniais described in this study. These compounds showed a significant dose-dependent inhibition effect on the proliferation of the parasites, with 50% inhibitory concentrations (IC50s) ranging from 1.23 to 25.05 μM against the promastigote stage and 0.5 to 0.7 μM against intracellular amastigotes. Also, a cytotoxicity assay was carried out to in order to evaluate the possible toxic effects of these compounds. Moreover, different assays were performed to determine the type of cell death induced after incubation with these compounds. The obtained results highlight the potential use of hexaazatrinaphthylene derivatives againstLeishmaniaspecies, and further studies should be undertaken to establish them as novel leishmanicidal therapeutic agents.

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