scholarly journals The in vitro toxicity of nitrile and epithionitrile derivatives of glucosinolates from rutabaga in human and bovine liver cells

2018 ◽  
Author(s):  
Ian Latimer ◽  
Mark Collett ◽  
Zoe Matthews ◽  
Brian Tapper ◽  
Belinda Cridge
Keyword(s):  
Cell Viability ◽  
No Reduction ◽  
Bovine Liver ◽  
Liver Cells ◽  
In Vitro Toxicity ◽  
Forage Crops ◽  
Previous Evidence ◽  
Toxic Event ◽  
Derivatives Of

Previous evidence suggests that select nitrile and epithionitrile derivatives of glucosinolates can cause liver disease in cows grazing on brassica forage crops. A toxic incidence in New Zealand in cattle grazing brassica led us to investigate the direct in vitro hepatotoxicity and possible inhibition of the ABCG2 transporter of five nitrile compounds. In this study, we investigated 1-cyano-2-hydroxy-3-butene (CHB, epithionitrile derivative of progoitrin), 1-cyano-2-hydroxy-3,4-epithiobutane (CHEB, nitrile derivative of progoitrin), 3-butenenitrile (nitrile from sinigrin), 4-pentenenitrile (nitrile from gluconapin), and 5-hexenenitrile (nitrile from glucobrassicanapin). Cell viability was assessed following 24- and 72-hr treatments with the 5 different compounds using the MTT assay (HepG2 cells and bovine primary liver cells). Additionally, ABCG2 transporter function was assessed. The results showed that none of the tested compounds caused cytotoxicity at concentrations up to 2 mM for 24hr. Over 72-hr the maximum concentration was 20 μM but no reduction in cell viability was observed. No inhibition of the ABCG2 transporter occured at concentrations up to 1 mM. Overall this study suggests that direct or secondary toxicity due to selected nitrile or epithionitrile derivatives of these glucosinolates was not the cause of the toxic event in cattle.

STUDIES ON OESTROGENS IN CATTLE

1960 ◽  
Vol XXXIV (I) ◽  
pp. 27-32 ◽  
Author(s):  
Stian Erichsen ◽  
Weiert Velle
Keyword(s):  
Bovine Liver ◽  
Liver Cells ◽  
Cell Sheets ◽  
Confluent Cell

ABSTRACT The metabolism of some oestrogenic hormones was studied in vitro by the use of cells grown on a medium free from blood. The methods used for the culture of cells from bovine testis, endometrium, amnion, and liver in confluent cell sheets on glass are described. The interconversion of oestrone and oestradiol-17β was demonstrated in the presence of cells from amnion, endometrium, and also from testicles of young calves and bulls. Only trace amounts of oestrone were found following incubations with oestradiol-17α in these tissues. Bovine liver cells grown in vitro showed a very poor capacity to bring about the interconversion mentioned above.

scholarly journals Huperzine A attenuates nonalcoholic fatty liver disease by regulating hepatocyte senescence and apoptosis: an in vitro study

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5145 ◽  
Author(s):  
Xiao-na Hu ◽  
Jiao-feng Wang ◽  
Yi-qin Huang ◽  
Zheng Wang ◽  
Fang-yuan Dong ◽  
...  
Keyword(s):  
Fatty Liver ◽  
Cell Viability ◽  
Accumulation Rate ◽  
Aging Effect ◽  
Liver Cells ◽  
Huperzine A ◽  
Inflammatory Factors ◽  
Fat Accumulation ◽  
Expression Levels

Objective This study was undertaken to detect if free fatty acids (FFA) induce hepatocyte senescence in L-02 cells and if huperzine A has an anti-aging effect in fatty liver cells. Methods L-02 cells were treated with a FFA mixture (oleate/palmitate, at 3:0, 2:1, 1:1, 1:2 and 0:3 ratios) at different concentrations. Cell viability and fat accumulation rate were assessed by a Cell Counting Kit 8 and Nile Red staining, respectively. The mixture with the highest cell viability and fat accumulation rate was selected to continue with the following experiment. The L-02 cells were divided into five groups, including the control group, FFA group, FFA + 0.1 μmol/L huperzine A (LH) group, FFA + 1.0 μmol/L huperzine A (MH) group and FFA + 10 μmol/L huperzine A (HH) group, and were cultured for 24 h. The expression of senescence-associated β-galactosidase (SA-β-gal) was detected by an SA-β-gal staining kit. The expression levels of aging genes were measured by qRT-PCR. The expression levels of apoptosis proteins were detected by a Western blot. ELISA kits were used to detect inflammatory factors and oxidative stress products. The expression of nuclear factor (NF-κB) and IκBα were detected by immunofluorescence. Results The FFA mixture (oleate/palmitate, at a 2:1 ratio) of 0.5 mmol/L had the highest cell viability and fat accumulation rate, which was preferable for establishing an in vitro fatty liver model. The expression of inflammatory factors (TNF-α and IL-6) and oxidants Malonaldehyde (MDA), 4-hydroxynonenal (HNE) and reactive oxygen species (ROS) also increased in the L-02 fatty liver cells. The expression levels of aging markers and aging genes, such as SA-β-gal, p16, p21, p53 and pRb, increased more in the L-02 fatty liver cells than in the L-02 cells. The total levels of the apoptosis-associated proteins Bcl2, Bax, Bax/Bcl-2, CyCt and cleaved caspase 9 were also upregulated in the L-02 fatty liver cells. All of the above genes and proteins were downregulated in the huperzine A and FFA co-treatment group. In the L-02 fatty liver cells, the expression of IκBα decreased, while the expression of NF-κB increased. After the huperzine A and FFA co-treatment, the expression of IκBα increased, while the expression of NF-κB decreased. Conclusion Fatty liver cells showed an obvious senescence and apoptosis phenomenon. Huperzine A suppressed hepatocyte senescence, and it might exert its anti-aging effect via the NF-κB pathway.

scholarly journals Comparative Proteomic Identification of Protein Disulphide Isomerase A6 Associated with Tert-Butylhydroperoxide-Induced Liver Injury in Rat Hepatocytes

2018 ◽  
Vol 45 (5) ◽  
pp. 1915-1926 ◽  
Author(s):  
Chien-Heng Shen ◽  
Shui-Yi Tung ◽  
Wen-Shih Huang ◽  
Kam-Fai Lee ◽  
Yung-Yu Hsieh ◽  
...  
Keyword(s):  
Liver Injury ◽  
Cell Viability ◽  
Er Stress ◽  
Primary Cultures ◽  
Liver Cells ◽  
Cytochrome C Release ◽  
Dapi Staining ◽  
Tert Butylhydroperoxide

Background/Aims: Oxidants are important human toxicants. They have been implicated in the occurrence and development of liver diseases. Increased intracellular tert-butylhydroperoxide (t-BHP) may be critical for oxidant toxicity, and is commonly used for evaluating mechanisms involving oxidative stress, but the method remains controversial. Methods: Primary cultures of hepatocytes as well as human Hep G2 and mouse FL83B liver cells were obtained. Cell viability was measured by annexin V–FITC/propidium iodide and DAPI staining to determine the effects of t-BHP treatment on acute liver injury. A proteomic assay provided information that was used to identify the differentially expressed proteins following t-BHP treatment; immunohistochemistry and western blotting were performed to detect the expression of PDIA6 activity in apoptotic and endoplasmic reticulum (ER) stress pathways. Results: Our results demonstrate that t-BHP treatment of liver cells increased cell cytotoxicity and the generation of reactive oxygen species. This treatment also increased the level of PDIA6; this was validated in vitro and in vivo based on a comparison of t-BHP-treated and -untreated groups. Treatment of mouse liver FL83B cells with t-BHP activated caspase 3, increased the expression of apoptotic molecules, caused cytochrome c release, and induced Bcl-2, Bax and IRE1α/TRAF2 complex formation. t-BHP-dependent induction of apoptosis was accompanied by sustained phosphorylation of the IRE1α/ASK1/JNK1/2/p38 pathways and PDIA6 expression. Furthermore, t-BHP induced liver FL83B cell viability and apoptosis by upregulating the levels of PDIA6; this process could be involved in the activation of the IRE1α/ASK1/JNK1/2/p38 signalling pathways. Conclusions: We conclude that t-BHP induced an apoptosis cascade and ER stress in hepatocytes by upregulation of PDIA6, providing a new mechanism underlying the effects of t-BHP on liver injury.

Long-term In Vitro Toxicity Models: Comparisons Between a Flow-cell Bioreactor, a Static-cell Bioreactor and Static Cell Cultures

2002 ◽  
Vol 30 (5) ◽  
pp. 515-523 ◽  
Author(s):  
Patricia Pazos ◽  
Salvador Fortaner ◽  
Pilar Prieto
Keyword(s):  
Cell Culture ◽  
Cell Viability ◽  
Flow Cell ◽  
Model Systems ◽  
In Vitro Toxicity ◽  
Protein Determination ◽  
Efficient System ◽  
Cytotoxicity Studies

In vitro long-term toxicity testing is becoming an important issue in the field of toxicology, and there is a need to develop new model systems that mimic human chronic exposure and its effects. The aim of this work was to test two long-term in vitro toxicity systems which are available, a flow-cell bioreactor (Tecnomouse) and a static cell bioreactor system (CELLine CL 6-well), and to compare them with the use of conventional cell culture flasks. A human cell line, Int 407, was exposed to cadmium chloride (CdCl2; 10–7–10–8M) for 4 weeks. Cell numbers and cell viabilities were determined by the trypan blue (TB) exclusion assay and from exclusion of propidium iodide (PI) as determined by flow cytometry; and cell viability and metabolic activity were determined by the MTT assay. In addition, total protein determination and cadmium uptake measurements were performed. The results obtained with TB and PI exclusion did not show clear differences in cell viability with increasing CdCl2 concentration. However, in the static cell-culture systems, an increase in MTT reduction was found at low concentrations of CdCl2. Expression of heat-shock protein (Hsp27 and Hsp70) increased differently, depending on the CdCl2 concentration applied and the system used. In summary, of the two bioreactors, the CELLine CL 6-well bioreactor was shown to be the more efficient system for performing long-term cytotoxicity studies. It is easy to handle, it permits the assessment of several endpoints, and sufficient replicates can be made available.

In vitro toxicity assessment of silver nanoparticles in Chang liver cells and J-774 macrophages

2010 ◽  
Vol 196 ◽  
pp. S284 ◽  
Author(s):  
L. Garza-Ocañas ◽  
M. Ramirez-Cabrera ◽  
M.T. Zanatta-Calderon ◽  
R. Lujan-Rangel ◽  
D.A. Ferrer ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Toxicity Assessment ◽  
Liver Cells ◽  
In Vitro Toxicity ◽  
In Vitro Toxicity Assessment ◽  
Chang Liver

In vitro toxicity of nanoparticles in BRL 3A rat liver cells

2005 ◽  
Vol 19 (7) ◽  
pp. 975-983 ◽  
Author(s):  
S.M. Hussain ◽  
K.L. Hess ◽  
J.M. Gearhart ◽  
K.T. Geiss ◽  
J.J. Schlager
Keyword(s):  
Rat Liver ◽  
Liver Cells ◽  
In Vitro Toxicity ◽  
Rat Liver Cells

scholarly journals In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus

2019 ◽  
Vol 5 (1) ◽  
Author(s):  
Murilo Batista Abud ◽  
Ricardo Noguera Louzada ◽  
David Leonardo Cruvinel Isaac ◽  
Leonardo Gomes Souza ◽  
Ricardo Gomes dos Reis ◽  
...  
Keyword(s):  
Cell Viability ◽  
Mtt Assay ◽  
Tunel Assay ◽  
In Vitro Toxicity ◽  
In Vivo Experiments ◽  
Significant Difference ◽  
The Difference ◽  
New Formulation

Abstract Background To evaluate the in vivo and in vitro toxicity of a new formulation of liposome-encapsulated sirolimus (LES). Methods In vitro experiments were done using ARPE-19 and HRP cells. An MTT assay was used to determine cell metabolic activity and a TUNEL assay for detecting DNA fragmentation. In vivo experiments were conducted on New Zealand albino rabbits that received intravitreal injections of empty liposomes (EL) or different concentrations of LES. Histopathological and immunohistochemical analyses were performed on the rabbit’s eyes following injection. Results Eighteen eyes of nine rabbits were used. MTT assay cell viability was 95.04% in group 1 (12.5 µL/mL LES). 92.95% in group 2 (25 µL/mL LES), 91.59% in group 3 (50 µL/mL LES), 98.09% in group 4 (12.5 µL/mL EL), 95.20% on group 5 (50 µL/mL EL), 98.53% in group 6 (50 µL/mL EL), and 2.84% on group 8 (50 µL/mL DMSO). There was no statistically significant difference among groups 1 to 7 in cell viability (p = 1.0), but the comparison of all groups with group 8 was significant (p < 0.0001). The TUNEL assay comparing two groups was not statistically significant from groups 1 to 7 (p = 1.0). The difference between groups 1 to 7 and group 8 (p < 0.0001) was significant. Histopathological changes were not found in any group. No activation of Müller cells was detected. Conclusion A novel formulation of LES delivered intravitreally did not cause in vitro toxicity, as evaluated by MTT and TUNEL assays, nor in vivo toxicity as evaluated by histopathology and immunohistochemistry in rabbit eyes.

Non-Clinical investigation of Tuberculosis Drugs - Conjugated Norbornene-Based Nanocarriers Toxic Impacts on Zebrafish

2021 ◽  
Vol 12 ◽  
Author(s):  
Thangammal Anju ◽  
Radhakrishnan Preetha ◽  
Raja Shunmugam ◽  
Shivshankar R Mane ◽  
Jesu Arockiaraj ◽  
...  
Keyword(s):  
Dna Damage ◽  
Cell Viability ◽  
Hepg2 Cells ◽  
In Vitro Study ◽  
Liver Cells ◽  
Vitro Study ◽  
Ros Generation ◽  
Human Liver Cell ◽  
Zebrafish Liver

INTRODUCTION: Rifampicin conjugated (R-CP), and rifampicin -isoniazid dual conjugated (RI-CP) norbornene-derived nanocarriers are newly designed for pH stimuli-responsive delivery of tuberculosis (TB) drugs. Its biosafety level is yet to be well established. OBJECTIVES: To assess the impacts of the nanocarriers on liver cells using zebrafish animal model and human liver cell line model (HepG2). METHODS: Initially, lethal dose concentration for the norbornene-derived nanocarrier systems in zebrafish was determined. The toxic effects were analysed at the sub-lethal drug concentration by histopathological study, total GSH level, gene expression and DNA damage in zebrafish liver cells. Fish erythrocyte nuclear abnormalities were also evaluated. Cell viability and oxidative stress level (ROS generation) after exposure to the nanoconjugates was determined using HepG2 cell in the in vitro study. RESULTS: In vivo studies of both R-CP and RI-CP showed 100% mortality at 96 hours for exposure concentration >100mg/l and showed toxic changes in zebrafish liver histology, GSH, and DNA damage levels. A noticeable upregulated PXR, CYP3A and cyp2p6 genes was observed in RI-CP exposure than in RIF or R-CP molecules. The in vitro study revealed a dose-dependent effect on cell viability and ROS generation for RIF, R-CP and RI-CP exposures in HepG2 cells. CONCLUSION: The current study reports that the rifampicin conjugated (R-CP) and rifampicin-isoniazid conjugated (RI-CP) norbornene derived nanocarriers exhibit enhanced toxic responses in both adult zebrafish and HepG2 cells. The pH-sensitive norbornene derived nanocarriers on conjugation with different drugs exhibited varied impacts on hepatic cells. Hence the present investigation recommends a complete metabolomics analysis and norbornene carrier-drug interaction study to be performed for each drug conjugated norbornene nanocarrier to ensure its biosafety.

23 Na Triple-quantum signal of in vitro human liver cells, liposomes, and nanoparticles: Cell viability assessment vs. separation of intra- and extracellular signal

2019 ◽  
Vol 50 (2) ◽  
pp. 435-444 ◽  
Author(s):  
Michaela A.U. Hoesl ◽  
Dennis Kleimaier ◽  
Ruomin Hu ◽  
Matthias Malzacher ◽  
Cordula Nies ◽  
...  
Keyword(s):  
Cell Viability ◽  
Human Liver ◽  
Liver Cells ◽  
Viability Assessment ◽  
Human Liver Cells ◽  
Extracellular Signal
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