scholarly journals Spasmolytic Effects of Aphanizomenon Flos Aquae (AFA) Extract on the Human Colon Contractility

Nutrients ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 3445
Author(s):  
Antonella Amato ◽  
Simona Terzo ◽  
Pierenrico Marchesa ◽  
Angela Maffongelli ◽  
Martina Martorana ◽  
...  
Keyword(s):  
Nervous System ◽  
Serotonin Receptor ◽  
Human Colon ◽  
Mechanical Activity ◽  
Pcr Analysis ◽  
Organ Bath ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Concentration Dependent Manner ◽  
Trace Amine

The blue-green algae Aphanizomenon flos aquae (AFA), rich in beneficial nutrients, exerts various beneficial effects, acting in different organs including the gut. Klamin® is an AFA extract particularly rich in β-PEA, a trace-amine considered a neuromodulator in the central nervous system. To date, it is not clear if β-PEA exerts a role in the enteric nervous system. The aims of the present study were to investigate the effects induced by Klamin® on the human distal colon mechanical activity, to analyze the mechanism of action, and to verify a β-PEA involvement. The organ bath technique, RT-PCR, and immunohistochemistry (IHC) were used. Klamin® reduced, in a concentration-dependent manner, the amplitude of the spontaneous contractions. EPPTB, a trace-amine receptor (TAAR1) antagonist, significantly antagonized the inhibitory effects of both Klamin® and exogenous β-PEA, suggesting a trace-amine involvement in the Klamin® effects. Accordingly, AphaMax®, an AFA extract containing lesser amount of β-PEA, failed to modify colon contractility. Moreover, the Klamin® effects were abolished by tetrodotoxin, a neural blocker, but not by L-NAME, a nitric oxide-synthase inhibitor. On the contrary methysergide, a serotonin receptor antagonist, significantly antagonized the Klamin® effects, as well as the contractility reduction induced by 5-HT. The RT-PCR analysis revealed TAAR1 gene expression in the colon and the IHC experiments showed that 5-HT-positive neurons are co-expressed with TAAR1 positive neurons. In conclusion, the results of this study suggest that Klamin® exerts spasmolytic effects in human colon contractility through β-PEA, that, by activating neural TAAR1, induce serotonin release from serotoninergic neurons of the myenteric plexus.

scholarly journals Exogenous glucagon-like peptide 1 reduces contractions in human colon circular muscle

2014 ◽  
Vol 221 (1) ◽  
pp. 29-37 ◽  
Author(s):  
Antonella Amato ◽  
Sara Baldassano ◽  
Rosa Liotta ◽  
Rosa Serio ◽  
Flavia Mulè
Keyword(s):  
Colonic Motility ◽  
Circular Muscle ◽  
Human Colon ◽  
Inhibitory Effect ◽  
Mechanical Activity ◽  
Organ Bath ◽  
Dependent Manner ◽  
Double Labeling ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1

Glucagon-like peptide 1 (GLP1) is a naturally occurring peptide secreted by intestinal L-cells. Though its primary function is to serve as an incretin, GLP1 reduces gastrointestinal motility. However, only a handful of animal studies have specifically evaluated the influence of GLP1 on colonic motility. Consequently, the aims of this study were to investigate the effects induced by exogenous GLP1, to analyze the mechanism of action, and to verify the presence of GLP1 receptors (GLP1Rs) in human colon circular muscular strips. Organ bath technique, RT-PCR, western blotting, and immunofluorescence were used. In human colon, exogenous GLP1 reduced, in a concentration-dependent manner, the amplitude of the spontaneous contractions without affecting the frequency and the resting basal tone. This inhibitory effect was significantly reduced by exendin (9–39), a GLP1R antagonist, which per se significantly increased the spontaneous mechanical activity. Moreover, it was abolished by tetrodotoxin, a neural blocker, or Nω-nitro-l-arginine – a blocker of neuronal nitric oxide synthase (nNOS). The biomolecular analysis revealed a genic and protein expression of the GLP1R in the human colon. The double-labeling experiments with anti-neurofilament or anti-nNOS showed, for the first time, that immunoreactivity for the GLP1R was expressed in nitrergic neurons of the myenteric plexus. In conclusion, the results of this study suggest that GLP1R is expressed in the human colon and, once activated by exogenous GLP1, mediates an inhibitory effect on large intestine motility through NO neural release.

scholarly journals Cytotoxic Effects of Flubendazole in Human Lung Cancer Cell Line A549

2020 ◽  
Vol 11 (4) ◽  
Author(s):  
Elham Hoveizi ◽  
Fatemeh Fakharzadeh Jahromi
Keyword(s):  
Lung Cancer ◽  
Cell Viability ◽  
Human Lung ◽  
A549 Cells ◽  
Beclin 1 ◽  
Human Lung Cancer ◽  
Pcr Analysis ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Acridine Orange Staining

Background: The development of effective anticancer drugs is a significant health issue. Previous studies showed that members of the benzimidazole family have anticancer effects on several cancers Objectives: The present study investigated the cytotoxic effect of flubendazole on A549 human lung cancer cells. Methods: The A549 cells were treated with flubendazole at 1, 2, 5, and 10 µM concentrations for three days. Cell viability was measured by the MTT assay and Acridine orange staining. Also, the expressions of P62 and Beclin -1 were analyzed by qRT-PCR analysis. Results: Cell viability of A549 cells, in a concentration-dependent manner, showed significant differences between the treatment and control groups, and the IC50 value was determined to be 2 µM. Also, flubendazole reduced the expression of P62 and increased the expression of Beclin 1 in treated cells. Conclusions: Flubendazole induces cell death in A549 cells in a dose and time-dependent manner and can offer new factors in lung cancer therapeutic strategies.

scholarly journals Relaxant effect of quercetin on rabbit isolated bladder smooth muscles contractions

2020 ◽  
Vol 10 (1) ◽  
pp. 61-67
Author(s):  
Hassan Sadraei ◽  
Sabihe Tabesh
Keyword(s):  
Smooth Muscles ◽  
Electrical Field Stimulation ◽  
Negative Control ◽  
Flavonoid Compound ◽  
Drug Candidate ◽  
Control Group ◽  
Organ Bath ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Relaxant Effect

Introduction: Quercetin is a flavonoid compound found in many medicinal plants. Antispasmodic effect of quercetin has been reported in ileum and uterus smooth muscles but not in bladder. Therefore, the objective of this research was to investigate relaxant effect of quercetin in rabbit isolated bladder. Methods: Male rabbit was asphyxiated with carbon dioxide and then sacrificed. The whole bladder was dissected out and placed in oxygenated Tyrode’s solution. Isolated bladder was cut into longitudinal strips and placed in an organ bath for contraction studies. Contractions were induced with KCl (20mM), acetylcholine (5μM) and electrical field stimulation (EFS). Full inhibitory concentration–response curve was constructed for quercetin following addition of above spasmogens. Quercetin was added into the organ bath with 2 fold increments in concentration until maximum response was achieved. Nifedipine was used as positive control group and equivalent volume of quercetin vehicle (water + DMSO) was used as negative control group.Results: Quercetin (4 μg/mL to 640 μg/mL) in a concentration dependent manner inhibited isolated bladder strips contracted by KCl (IC50=159±25 μg/mL), acetylcholine (IC50=43±9.1 μg/mL) and EFS (IC50=38±9.3 μg/mL). In the highest used concentration, quercetin completely removed contractile responses to KCl, acetylcholine and electrical filed stimulation (EFS). Nifedipine totally inhibited KCl response (IC50=115±36 ng/mL) but only partially inhibited acetylcholine and EFS responses. Conclusion: These results confirm the relaxant effect of quercetin on rabbit bladder and if similar effects are seen in human studies, then quercetin would be a suitable drug candidate to be investigated for bladder incontinence.

Naked Poly(amidoamine) Dendrimer Nanoparticles Exhibit Intrinsic Embryotoxicity During the Early Stages of Normal Development

2020 ◽  
Vol 16 (10) ◽  
pp. 1454-1462
Author(s):  
Hadeel Kheraldine ◽  
Ishita Gupta ◽  
Hashim Alhussain ◽  
Ayesha Jabeen ◽  
Saghir Akhtar ◽  
...  
Keyword(s):  
Normal Development ◽  
Chorioallantoic Membrane ◽  
Pamam Dendrimers ◽  
Pcr Analysis ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Future Studies ◽  
Early Stages ◽  
Expression Of Genes ◽  
The Impact

To investigate the impact of poly(amidoamine) dendrimers (PAMAMs) in the embryo, we explored the outcome of different generations (G4 and G6) on the early stages of embryogenesis using the chicken embryo as a model. We also monitored their effect on angiogenesis in the chorioallantoic membrane (CAM). Our data revealed that cationic PAMAMs provoke substantial embryotoxicity, as they significantly induce death (up to 50%, p < 0 05) and inhibit angiogenesis of the CAM (up to 30%, p < 0 05) in a generation-dependent manner in comparison to controls and other types of PAMAMs (anionic and neutral). Moreover, cationic PAMAMs alter the expression of genes related to cell survival, cell cycle, proliferation, transcription factor, apoptosis, and angiogenesis, as shown by RT-PCR analysis. Our data suggest that PAMAM dendrimers exhibit intrinsic toxicity in embryos at the early stages and inhibits angiogenesis of the CAM. Thus, future studies are necessary to illustrate the exact mechanism of PAMAM dendrimers in embryotoxicity.

scholarly journals Calcitonin Induces IL-6 Production via Both PKA and PKC Pathways in the Pituitary Folliculo-Stellate Cell Line

Endocrinology ◽  
2001 ◽  
Vol 142 (8) ◽  
pp. 3563-3569 ◽  
Author(s):  
Yoshimitsu Kiriyama ◽  
Hiroyuki Tsuchiya ◽  
Takeshi Murakami ◽  
Kumi Satoh ◽  
Yukiko Tokumitsu
Keyword(s):  
Cell Line ◽  
Stellate Cell ◽  
Fold Increase ◽  
Receptor Subtype ◽  
Dependent Manner ◽  
Calcitonin Receptor ◽  
Rt Pcr ◽  
Concentration Dependent Manner ◽  
Mouse Pituitary ◽  
Inhibitor Treatment

Abstract It has been demonstrated that calcitonin-binding sites are present in a variety of tissue types, including in the pituitary gland. Interleukin-6 (IL-6) is also produced in the pituitary and it regulates the secretion of various hormones. In this study, we examined the expression of the calcitonin receptor and the mechanism of IL-6 production induced by calcitonin in the pituitary folliculo-stellate cell line (TtT/GF). The mRNA of calcitonin receptor subtype C1a, but not that of C1b, was detected by RT-PCR in TtT/GF cells and in the normal mouse pituitary. Calcitonin increased cAMP accumulation and IL-6 production in a concentration-dependent manner in TtT/GF cells. As calcitonin activates the PKA and PKC pathways, we investigated the contributions of PKA and PKC to IL-6 production. IL-6 production was only slightly increased by either 8-bromo-cAMP (1 mm) or phorbol 12-myristate 13-acetate (100 nm) alone. However, IL-6 was synergistically induced in the presence of both 8-bromo-cAMP (1 mm) and phorbol 12myristate 13-acetate (100 nm). Furthermore, calcitonin-induced IL-6 production was completely suppressed by H-89 (PKA inhibitor) or GF109203X (PKC inhibitor), indicating that the activation of both PKA and PKC is necessary for calcitonin-induced IL-6 production. On the other hand, pertussis toxin (Gi/Go signaling inhibitor) treatment achieved an approximately 9-fold increase in calcitonin-induced IL-6 production. These results show that calcitonin-stimulated IL-6 production is mediated via both PKA- and PKC-signaling pathways, whereas calcitonin also suppresses IL-6 production by activating Gi/Go proteins in folliculo-stellate cells.

Effects of Buthus martensii Karsch Venom on Cell Proliferation and Cytotoxicity in Hela Cells

2011 ◽  
Vol 345 ◽  
pp. 393-398 ◽  
Author(s):  
Zhi Wang ◽  
Wen Hui Fu ◽  
Xiang Yang Lu ◽  
Guang Xian Cai
Keyword(s):  
Hela Cell ◽  
Hela Cells ◽  
Inhibitory Concentration ◽  
Scorpion Venom ◽  
Dependent Manner ◽  
Hela Cell Line ◽  
Rt Pcr ◽  
Concentration Dependent Manner ◽  
Significant Difference ◽  
Apoptosis And Necrosis

This paper focuses on the effect of the venom of the scorpion Buthus martensii on the proliferation of human cervical carcinoma Hela cell line and the related molecular mechanism. MTT test showed that the scorpion venom inhibited proliferation of Hela cells in time-dependent and concentration-dependent manner with 50% inhibitory concentration (IC50) of 34.5 μg/mL(48 h). By using flow cytometry, it was found that the scorpion venom could induce apoptosis and necrosis in Hela cells. RT-PCR and Western blot indicated there were obviously up-regulated in the expressions of p21 protein but the expression of p21 mRNA showed no significant difference in the Hela cell by the scorpion venom. These results suggest that the possible mechanism of the scorpion venom is to activate the expressions of p21 protein and to cause Hela cell apoptosis.

scholarly journals Inhibition of human ornithine decarboxylase activity by enantiomers of difluoromethylornithine

2003 ◽  
Vol 375 (2) ◽  
pp. 465-470 ◽  
Author(s):  
Ning QU ◽  
Natalia A. IGNATENKO ◽  
Phillip YAMAUCHI ◽  
David E. STRINGER ◽  
Corey LEVENSON ◽  
...  
Keyword(s):  
Ornithine Decarboxylase ◽  
Enzyme Inhibitor ◽  
Human Colon ◽  
Common Mechanism ◽  
Decarboxylase Activity ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Polyamine Biosynthesis ◽  
Irreversible Reaction ◽  
Kd Values

Racemic difluoromethylornithine (d/l-DFMO) is an inhibitor of ODC (ornithine decarboxylase), the first enzyme in eukaryotic polyamine biosynthesis. d/l-DFMO is an effective anti-parasitic agent and inhibitor of mammalian cell growth and development. Purified human ODC-catalysed ornithine decarboxylation is highly stereospecific. However, both DFMO enantiomers suppressed ODC activity in a time- and concentration-dependent manner. ODC activity failed to recover after treatment with either l- or d-DFMO and dialysis to remove free inhibitor. The inhibitor dissociation constant (KD) values for the formation of enzyme–inhibitor complexes were 28.3±3.4, 1.3±0.3 and 2.2±0.4 μM respectively for d-, l- and d/l-DFMO. The differences in these KD values were statistically significant (P<0.05). The inhibitor inactivation constants (Kinact) for the irreversible step were 0.25±0.03, 0.15±0.03 and 0.15±0.03 min−1 respectively for d-, l- and d/l-DFMO. These latter values were not statistically significantly different (P>0.1). d-DFMO was a more potent inhibitor (IC50~7.5 μM) when compared with d-ornithine (IC50~1.5 mM) of ODC-catalysed l-ornithine decarboxylation. Treatment of human colon tumour-derived HCT116 cells with either l- or d-DFMO decreased the cellular polyamine contents in a concentration-dependent manner. These results show that both enantiomers of DFMO irreversibly inactivate ODC and suggest that this inactivation occurs by a common mechanism. Both enantiomers form enzyme–inhibitor complexes with ODC, but the probability of formation of these complexes is 20 times greater for l-DFMO when compared with d-DFMO. The rate of the irreversible reaction in ODC inactivation is similar for the l- and d-enantiomer. This unexpected similarity between DFMO enantiomers, in contrast with the high degree of stereospecificity of the substrate ornithine, appears to be due to the α-substituent of the inhibitor. The d-enantiomer may have advantages, such as decreased normal tissue toxicity, over l- or d/l-DFMO in some clinical applications.

Effects of plant lectin and extracts on adhesion molecules of endothelial progenitors

2011 ◽  
Vol 6 (3) ◽  
pp. 330-341 ◽  
Author(s):  
Florin Iordache ◽  
Iordache Carmen ◽  
Pop Aneta ◽  
Marilena Lupu ◽  
Eugen Andrei ◽  
...  
Keyword(s):  
Gene Expression ◽  
Adhesion Molecules ◽  
Adult Stem Cells ◽  
Mononuclear Cells ◽  
Vascular Tissue ◽  
Dependent Manner ◽  
Wound Healing Assay ◽  
Rt Pcr ◽  
Calendula Officinalis ◽  
Concentration Dependent Manner

AbstractPromise of cell therapy has advanced the use of adult stem cells towards the development of novel approaches to promote regeneration of injured endothelium. The aim of this study was to stimulate endothelial progenitor cells (EPCs) with lectin isolated from Solanum tuberosum (potato) shoot and Calendula officinalis (marigold) extracts, in order to increase EPCs proliferation and gene expression of molecules with roles in chemotaxis and adhesion for a better attachment to injured vascular tissue. EPCs were differentiated from umbilical cord blood-derived mononuclear cells and characterized by light microscopy, flow cytometry, and vascular tube-like structures formation on Matrigel. Cell proliferation was determined by MTS assay, and gene expression of molecules involved in EPCs adhesion (VCAM-1, VE-cadherin, ICAM-1, PECAM-1, P-selectin) and chemotaxis was determined (CXCR4, Tie-2) by RT-PCR. For the assessment of cell motility, wound-healing assay was employed. Both potato shoot lectin and marigold extracts stimulated EPCs proliferation in a concentration dependent manner and were able to increase expression of adhesion and chemotactic molecules. Marigold flower extract proved to be more efficient. This study demonstrates the usefulness of potato lectin and marigold extracts to increase EPCs proliferation and modulate gene expression of chemotactic and adhesion molecules, which may facilitate EPCs attachment to injured endothelium.

scholarly journals Cannabinoid CB2 receptors in the enteric nervous system modulate gastrointestinal contractility in lipopolysaccharide-treated rats

2008 ◽  
Vol 295 (1) ◽  
pp. G78-G87 ◽  
Author(s):  
Marnie Duncan ◽  
Abdeslam Mouihate ◽  
Ken Mackie ◽  
Catherine M. Keenan ◽  
Nancy E. Buckley ◽  
...  
Keyword(s):  
Nervous System ◽  
Enteric Nervous System ◽  
Myenteric Plexus ◽  
Electrical Field Stimulation ◽  
Receptor Expression ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Receptor Agonists ◽  
Lps Treatment

Enhanced intestinal transit due to lipopolysaccharide (LPS) is reversed by cannabinoid (CB)2 receptor agonists in vivo, but the site and mechanism of action are unknown. We have tested the hypothesis that CB2 receptors are expressed in the enteric nervous system and are activated in pathophysiological conditions. Tissues from either saline- or LPS-treated (2 h; 65 μg/kg ip) rats were processed for RT-PCR, Western blotting, and immunohistochemistry or were mounted in organ baths where electrical field stimulation was applied in the presence or absence of CB receptor agonists. Whereas the CB2 receptor agonist JWH133 did not affect the electrically evoked twitch response of the ileum under basal conditions, in the LPS-treated tissues JWH133 was able to reduce the enhanced contractile response in a concentration-dependent manner. Rat ileum expressed CB2 receptor mRNA and protein under physiological conditions, and this expression was not affected by LPS treatment. In the myenteric plexus, CB2 receptors were expressed on the majority of neurons, although not on those expressing nitric oxide synthase. LPS did not alter the distribution of CB2 receptor expression in the myenteric plexus. In vivo LPS treatment significantly increased Fos expression in both enteric glia and neurons. This enhanced expression was significantly attenuated by JWH133, whose action was reversed by the CB2 receptor antagonist AM630. Taking these facts together, we conclude that activation of CB2 receptors in the enteric nervous system of the gastrointestinal tract dampens endotoxin-induced enhanced intestinal contractility.

scholarly journals Rhododendron molle G. Don Extract Induces Apoptosis and Inhibits Migration in Human Colorectal Cancer Cells and Potential Anticancer Components Analysis

Molecules ◽  
2021 ◽  
Vol 26 (10) ◽  
pp. 2990
Author(s):  
Luye Zong ◽  
Yan Yang ◽  
Jin Zhang ◽  
Liangfang Dai ◽  
Yuqiang Luo ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
S Phase ◽  
Pcr Analysis ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Qrt Pcr ◽  
Medicinal Value ◽  
Colorectal Cancer Cells ◽  
And Migration ◽  
Ht 29

Rhododendron molle G. Don is one example of traditional Chinese medicine with important medicinal value. In this study, the effects of methanol extract of R. molle leaves (RLE) on colorectal cancer HT-29 cells and its potential molecular mechanism were investigated. MTT analysis showed that RLE could significantly inhibit the cell viability and migration of HT-29 cells in a concentration-dependent manner. Cell cycle analyses via flow cytometer suggested that RLE induced DNA fragmentation, indicative of apoptosis, and arrest at the S phase in HT-29 cells. Quantitative real-time PCR (qRT-PCR) analysis showed that RLE could upregulate the mRNA expression of p53 and p21 in HT-29 cells, which would result in HT-29 cells being blocked in S phase. Meanwhile, RLE could upregulate the expression of Bax, and downregulate the expression of Bcl-2, which would induce cell apoptosis. Further western blot analysis showed that the protein expression changes of Bax and P53 were basically consistent with the results of qRT-PCR. In addition, GC-MS analysis detected 17 potential anticancer components in R. molle. These results indicate that R. molle has significant anticancer activity, which provides some useful information for further study and clinical application for R. molle.

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