The effect of Apium Graveolens L., Levisticum Officinale and Calendula Officinalis L. on cell viability, membrane integrity, steroidogenesis, and intercellular communication in mice Leydig cells in vitro

Several plants have the potential to protect essential reproductive processes such as spermatogenesis or steroidogenesis, however, effective concentrations and main mechanisms of action are still unknown. This in vitro study was aimed to assess the effects of Apium graveolens L., Levisticum officinale, and Calendula officinalis L. extracts on the structural integrity, functional activity and gap junctional intercellular communication (GJIC) in mice Leydig cells. TM3 cells were grown in the presence of experimental extracts (37.5, 75, 150 and 300 µg/ml) for 24 h. For the present study, high-performance liquid chromatography analysis was used to quantify flavonoids or phenolic acids. Subsequently, Leydig cell viability was assessed by alamarBlue assay, while the cell membrane integrity was detected by 5 carboxyfluorescein diacetate-acetoxymethyl ester. The level of steroid hormones production was determined by enzyme-linked immunosorbent assay. Additionally, GJIC was assessed by scalpel loading/dye transfer assay. According to our results, Apium graveolens L. significantly increased the viability and cell membrane integrity at 75 µg/ml (109.0±4.3 %) followed by a decline at 300 µg/ml (89.4±2.3 %). In case of Levisticum officinale and Calendula officinalis L. was observed significant decrease at 150 µg/ml (88.8±11.66 %, 87.4±6.0 %) and 300 µg/ml (86.2±9.3 %, 84.1±4.6 %). Furthermore, Apium graveolens L. significantly increased the progesterone and testosterone production (75 and 150 µg/ml) however, Levisticum officinale and Calendula officinalis L. significantly reduced steroid hormones synthesis at 150 and 300 µg/ml. Finally, the disturbance of GJIC was significantly affected at 300 µg/ml of Levisticum officinale (82.5±7.7 %) and Calendula officinalis L. (79.8±7.0 %). The balanced concentration ratio may support the Leydig cell function, steroidogenesis as well as all essential parameters that may significantly improve reproductive functions.

Download Full-text

Assessment of changes in sperm cell membrane integrity occurring during the storage of semen from genetically different males using two diagnostic methods

Canadian Journal of Animal Science ◽

10.4141/cjas2013-095 ◽

2014 ◽

Vol 94 (4) ◽

pp. 601-606 ◽

Cited By ~ 2

Author(s):

Anna Wysokińska ◽

Stanislaw Kondracki

Keyword(s):

Cell Membrane ◽

Sperm Cell ◽

Cell Membranes ◽

Intact Cell ◽

Membrane Integrity ◽

Diagnostic Methods ◽

Cell Membrane Integrity ◽

Staining Methods ◽

Breed Crosses

Wysokińska, A. and Kondracki, S. 2014. Assessment of changes in sperm cell membrane integrity occurring during the storage of semen from genetically different males using two diagnostic methods. Can. J. Anim. Sci. 94: 601–606. The present study was carried out to assess changes in sperm cell membrane integrity occurring during the storage of semen collected from genetically different domestic male pigs. The study was aimed at assessing differences in the course of changes in the integrity of cell membranes in spermatozoa produced by males with different degrees of genetic diversity (pure-bred males, two-breed hybrids and multi-breed crosses) and testing the usefulness of two methods of sperm cell membrane integrity evaluation, based on material collected from genetically different males. The experiments were conducted on 56 ejaculates collected from 28 domestic male pigs. The examination of sperm cell membrane integrity was performed three times for each ejaculate, i.e., after 1 h, after 24 h and after 48 h from collection. The preparations for analysing cell membrane integrity were made using two methods: the SYBR 14/PI method and the eosin–nigrosin method. It was found that both SYBR 14/PI and eosin–nigrosin staining methods make it possible to successfully assess the integrity of the plasma membrane of domestic pig sperm cells under in vitro conditions. Hybrid pig spermatozoa, especially those from multi-breed crosses, better retain the integrity of their plasmalemmas than the spermatozoa of pure-bred boars. The ejaculates of Hypor cross-breed boars assessed after 1, 24 and 48 h of storage contain more spermatozoa with intact cell membranes than the ejaculates of pure-bred Duroc and Pietrain boars. The ejaculates of Hypor boars also show fewer decaying spermatozoa than those produced by pure-bred boars.

Download Full-text

Altering the Proclivity towards Daptomycin Resistance in Methicillin-Resistant Staphylococcus aureus Using Combinations with Other Antibiotics

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00502-12 ◽

2012 ◽

Vol 56 (10) ◽

pp. 5046-5053 ◽

Cited By ~ 40

Author(s):

Andrew D. Berti ◽

Justine E. Wergin ◽

Gary G. Girdaukas ◽

Scott J. Hetzel ◽

George Sakoulas ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Cell Wall ◽

Cell Membrane ◽

Membrane Integrity ◽

Wall Thickening ◽

Methicillin Resistant ◽

Content Type ◽

Cell Membrane Integrity ◽

Cell Membrane Fluidity

ABSTRACTDaptomycin (DAP) is increasingly used as a part of combination therapy, particularly in complex methicillin-resistantStaphylococcus aureus(MRSA) infections. While multiple studies have reported the potential for synergy between DAP and adjunctive anti-infectives, few have examined the influence of adjunctive therapy on the emergence of DAP resistance. This study examined eight adjunctive antimicrobial combinations with DAPin vitroand the emergence of DAP resistance over time (up to 4 weeks) using clinical isolates of DAP-susceptible MRSA (MIC, 0.5 μg/ml) in which DAP resistance subsequently developed during patient therapy (MIC, 3 μg/ml). In addition to DAP susceptibility testing, selected strains were examined for phenotypic changes associated with DAP resistance, including changes to cell wall thickness (CWT) and cell membrane alterations. The addition of either oxacillin or clarithromycin in medium containing DAP significantly inhibited the development of DAP resistance through the entirety of the 4-week exposure (10- to 32-fold MIC reduction from that of DAP alone). Combinations with rifampin or fosfomycin were effective in delaying the emergence of DAP resistance through the end of week one only (week one MIC, 0.5 μg/ml; week four MIC, 24 μg/ml). Cell wall thickening was observed for all antibiotic combinations regardless of their effect on the DAP MIC (14 to 70% increase in CWT), while changes in cell membrane fluidity were variable and treatment dependent. DAP showed reduced activity against strains with DAP MICs of 1 to 12 μg/ml, but cell membrane integrity was still disrupted at concentrations achieved with doses greater than 10 mg/kg of body weight. The emergence of DAP resistance in MRSA is strongly influenced by the presence of subinhibitory concentrations of adjunctive antimicrobials. These data suggest that combining DAP with oxacillin or clarithromycin may delay the development of DAP resistance in cases requiring prolonged antibiotic therapy.

Download Full-text

Red blood cell membrane integrity in primary open angle glaucoma: ex vivo and in vitro studies

Eye ◽

10.1038/eye.1999.18 ◽

1999 ◽

Vol 13 (1) ◽

pp. 101-103 ◽

Cited By ~ 23

Author(s):

L Zabala ◽

C Saldanha ◽

J Martins E Silva ◽

P Souza-Ramalho

Keyword(s):

Cell Membrane ◽

Blood Cell ◽

Red Blood Cell ◽

Primary Open Angle Glaucoma ◽

Ex Vivo ◽

Open Angle Glaucoma ◽

Membrane Integrity ◽

Open Angle ◽

Cell Membrane Integrity

Download Full-text

Carvacrol exhibits rapid bactericidal activity against Streptococcus pyogenes through cell membrane damage

Scientific Reports ◽

10.1038/s41598-020-79713-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Niluni M. Wijesundara ◽

Song F. Lee ◽

Zhenyu Cheng ◽

Ross Davidson ◽

H. P. Vasantha Rupasinghe

Keyword(s):

Cell Membrane ◽

Streptococcus Pyogenes ◽

Bactericidal Activity ◽

Membrane Damage ◽

Membrane Integrity ◽

Streptococcal Pharyngitis ◽

Cell Membrane Integrity ◽

Cell Membrane Damage ◽

Antibiotic Development

AbstractStreptococcus pyogenes is an important human pathogen worldwide. The identification of natural antibacterial phytochemicals has renewed interest due to the current scarcity of antibiotic development. Carvacrol is a monoterpenoid found in herbs. We evaluated carvacrol alone and combined with selected antibiotics against four strains of S. pyogenes in vitro. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of carvacrol against S. pyogenes were 125 µg/mL (0.53 mM) and 250 µg/mL (1.05 mM), respectively. Kill curve results showed that carvacrol exhibits instantaneous bactericidal activity against S. pyogenes. We also demonstrated the potential mechanism of action of carvacrol through compromising the cell membrane integrity. Carvacrol induced membrane integrity changes leading to leakage of cytoplasmic content such as lactate dehydrogenase enzymes and nucleic acids. We further confirmed dose-dependent rupturing of cells and cell deaths using transmission electron microscopy. The chequerboard assay results showed that carvacrol possesses an additive-synergistic effect with clindamycin or penicillin. Carvacrol alone, combined with clindamycin or penicillin, can be used as a safe and efficacious natural health product for managing streptococcal pharyngitis.

Download Full-text

In vitro Studies on PMIM-lndependent Endothelial Cell Damage in Trauma: Decrease of PMN-Endothelial Cell Adherence by Fibrinogen Degradation Products and Disturbance of Endothelial Cell Membrane Integrity by Trauma Serum

European Surgical Research ◽

10.1159/000129261 ◽

1993 ◽

Vol 25 (2) ◽

pp. 83-90

Author(s):

P. Vogel ◽

J.v.d. Beek ◽

K. Marohl ◽

E.G. Fischer ◽

C.J. Kirkpatrick

Keyword(s):

Endothelial Cell ◽

Cell Membrane ◽

Degradation Products ◽

Cell Damage ◽

Membrane Integrity ◽

In Vitro Studies ◽

Cell Adherence ◽

Endothelial Cell Damage ◽

Cell Membrane Integrity

Download Full-text

2-Phenylethyl Isothiocyanate Exerts Antifungal Activity against Alternaria alternata by Affecting Membrane Integrity and Mycotoxin Production

Toxins ◽

10.3390/toxins12020124 ◽

2020 ◽

Vol 12 (2) ◽

pp. 124

Author(s):

Miao Zhang ◽

Yongcai Li ◽

Yang Bi ◽

Tiaolan Wang ◽

Yupeng Dong ◽

...

Keyword(s):

Cell Membrane ◽

Alternaria Alternata ◽

Membrane Integrity ◽

Black Spot ◽

Cell Membrane Permeability ◽

Pear Fruit ◽

Cell Membrane Integrity ◽

Inhibitory Mechanisms ◽

Phenylethyl Isothiocyanate

Black spot caused by Alternaria alternata is one of the important diseases of pear fruit during storage. Isothiocyanates are known as being strong antifungal compounds in vitro against different fungi. The aim of this study was to assess the antifungal effects of the volatile compound 2-phenylethyl isothiocyanate (2-PEITC) against A. alternata in vitro and in pear fruit, and to explore the underlying inhibitory mechanisms. The in vitro results showed that 2-PEITC significantly inhibited spore germination and mycelial growth of A. alternata—the inhibitory effects showed a dose-dependent pattern and the minimum inhibitory concentration (MIC) was 1.22 mM. The development of black spot rot on the pear fruit inoculated with A. alternata was also significantly decreased by 2-PEITC fumigation. At 1.22 mM concentration, the lesion diameter was only 39% of that in the control fruit at 7 days after inoculation. Further results of the leakage of electrolyte, increase of intracellular OD260, and propidium iodide (PI) staining proved that 2-PEITC broke cell membrane permeability of A. alternata. Moreover, 2-PEITC treatment significantly decreased alternariol (AOH), alternariolmonomethyl ether (AME), altenuene (ALT), and tentoxin (TEN) contents of A. alternata. Taken together, these data suggest that the mechanisms underlying the antifungal effect of 2-PEITC against A. alternata might be via reduction in toxin content and breakdown of cell membrane integrity.

Download Full-text

Investigation of the properties and effects of salvia officinalis l. on the viability, steroidogenesis and Reactive Oxygen Species (ROS) production in TM3 leydig cells in vitro

Physiological Research ◽

10.33549/physiolres.934457 ◽

2020 ◽

pp. 661-673

Author(s):

T Jambor ◽

J Arvay ◽

E Ivanisova ◽

E Tvrda ◽

A Kovacik ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Cell Viability ◽

Leydig Cell ◽

Leydig Cells ◽

Membrane Integrity ◽

Salvia Officinalis ◽

Scavenging Activity ◽

Oxygen Species ◽

Salvia Officinalis L

The aim of our study was to reveal the in vitro effects of Salvia officinalis L. (37.5, 75, 150, 200, 250, 300 and 600 µg/ml) extract on the TM3 Leydig cell viability, membrane integrity, steroidogenesis and reactive oxygen species production after 24 h and 48 h cultivation. For the present study, the extract prepared from Salvia officinalis L. leaves was analysed by high performance liquid chromatography (HPLC) for selected flavonoids and phenolic acids followed by a determination of its free radicals scavenging activity (DPPH). Furthermore, Leydig cell viability was assessed by the mitochondrial toxicity assay (MTT), while the membrane integrity was evaluated by 5- carboxyfluorescein diacetate-acetoxymethyl ester (5-CFDA-AM). The level of steroid hormones was performed by enzyme-linked immunosorbent assay (ELISA) from the culture media, while the superoxide radical generation was measured by the nitroblue tetrazolium chloride (NBT) assay. The results show that experimental concentrations did not damage the cell membrane integrity and viability when present at below 300 µg/ml, it was only at 600 µg/ml that a significant (P<0.05) cell viability decline was observed after a 48 h cultivation. A significant (P<0.05) stimulation of testosterone secretion was recorded at 250 µg/ml for 24 h, while the prolonged cultivation time significantly (P<0.05) increased the testosterone and progesterone production at 150, 200, 250 and 300 µg/ml. Furthermore, none of the selected doses exhibited significant ROS-promoting effects however, the highest dose of Salvia initiated the free radical scavenging activity in cultured mice Leydig cells.

Download Full-text