scholarly journals Mitochondrial membrane potential played crucial roles in the accumulation of berberine in HepG2 cells

2019 ◽  
Vol 39 (4) ◽  
Author(s):  
Qiao Li ◽  
Ting Zhou ◽  
Chang Liu ◽  
Xiao-Yu Wang ◽  
Ji-Quan Zhang ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Cell Membrane ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Cell Membrane Potential ◽  
Cell Organelles ◽  
Chromatography Method

Abstract Berberine is a natural alkaloid that has antineoplastic effects. However, in hepatoma cells like HepG2, the expressions of uptake transporters are minimal but efflux transporters are relatively high. Hence, how berberine enters and reaches a cytocidal concentration remains to be elucidated. In the present study, we revealed the accumulation mechanism of berberine in HepG2 cells. Cell organelles were isolated based on differential centrifugation; berberine concentration was measured using a liquid chromatography-tandem mass chromatography method or flow cytometry. Subcellular distribution of berberine was observed using a laser scanning confocal microscopy. The results showed that berberine was concentration-, temperature-, and time-dependently taken up and accumulated in HepG2 cells. Membrane drug transporters and cell membrane potential had limited effects in berberine uptake. However, qualitative and quantitative studies showed that berberine was enriched in the mitochondria; inhibition of mitochondrial membrane potential (MMP) by carbonyl cyanide 3-chlorophenylhydrazone (CCCP) significantly decreased the intracellular berberine by up to 70%. More importantly, MMP not only significantly enhanced berberine uptake driven by cell membrane potential (P<0.01) but also inhibited p-glycoprotein (P-gp)-mediated berberine efflux (P<0.01). In brief, our results for the first time showed that MMP played crucial roles in berberine accumulation in HepG2 cells.

scholarly journals Does Cryopreservation of Ovarian Tissue Affect the Distribution and Function of Germinal Vesicle Oocytes Mitochondria?

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Mojdeh Salehnia ◽  
Virpi Töhönen ◽  
Saeed Zavareh ◽  
Jose Inzunza
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Laser Scanning ◽  
Ovarian Tissue ◽  
Germinal Vesicle ◽  
Free Calcium ◽  
Atp Content ◽  
Fertilization Rates

The aim of this study was to evaluate mitochondrial alteration and ATP content of germinal vesicle (GV) oocytes isolated from fresh and vitrified ovaries. After superovulation, the ovaries from adult mice were collected and divided into control and vitrified groups. GV oocytes were isolated mechanically from each group. Half were cultured for 24 hours and their maturation was assessed. Metaphase II oocytes were collected and submitted toin vitrofertilization and their fertilization rates and development to the blastocyst stage were evaluated. In the remaining GV oocytes, ATP levels were quantified, and mitochondrial distribution, mitochondrial membrane potential, and intracellular free calcium were detected with rhodamine 123, JC-1 and Flou-4 AM staining, using laser-scanning confocal microscopy. Maturation and fertilization rates of GV oocytes and the developmental rates of subsequent embryos were significantly lower in vitrified samples (P<0.05). The ATP content and Ca2+levels differed significantly in fresh and vitrified GV oocytes (P<0.05). Most mitochondria were seen as large and homogenous aggregates (66.6%) in fresh GV oocytes compared to vitrified oocytes (50%). No significant differences in mitochondrial membrane potential were found between the groups. The lower maturation and fertilization rates of GV oocytes from vitrified ovaries may be due to changes in their mitochondrial function and distribution.

Lysosome Inhibitors Enhance the Chemotherapeutic Activity of Doxorubicin in HepG2 Cells

Chemotherapy ◽  
2016 ◽  
Vol 62 (2) ◽  
pp. 85-93 ◽  
Author(s):  
Yuyin Li ◽  
Yuejun Sun ◽  
Lifang Jing ◽  
Jianjun Wang ◽  
Yali Yan ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Inhibitory Effect ◽  
Membrane Permeabilization ◽  
Lysosomal Membrane ◽  
Bafilomycin A1 ◽  
Lysosomal Membrane Permeabilization ◽  
Proliferative Ability

The lysosome inhibitors bafilomycin A1 and chloroquine have both lysosomotropic properties and autophagy inhibition ability, and are promising clinical agents to be used in combination with anticancer drugs. In order to investigate this combination effect, HepG2 cells were treated with bafilomycin A1, chloroquine, or/and doxorubicin, and their proliferative ability, induction of apoptosis, and the changes of lysosomal membrane permeabilization and mitochondrial membrane potential were studied. The results demonstrate that treatment with bafilomycin A1 or chloroquine alone at a relatively low concentration promotes the inhibitory effect of doxorubicin on cell growth and apoptosis. Further studies reveal that bafilomycin A1 and chloroquine promote lysosomal membrane permeabilization and the reduction of mitochondrial membrane potential induced by doxorubicin. Our findings suggest that bafilomycin A1 and chloroquine potentiate the anticancer effect of doxorubicin in hepatic cancer cells and that supplementation of conventional chemotherapy with lysosome inhibitors may provide a more efficient anticancer therapy.

Study on Capparis spionosa L. Polysaccharide (CSPS) Induced HepG2 Apoptosis by Controlling Ca2+ Path

2011 ◽  
Vol 282-283 ◽  
pp. 203-208 ◽  
Author(s):  
Yu Bin Ji ◽  
Fang Dong ◽  
Shi Yong Gao ◽  
Miao Yu
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Hepatoma Cell ◽  
Scanning Microscope ◽  
Human Hepatoma Cell ◽  
Bax Protein ◽  
Confocal Scanning ◽  
Labeling Method

To investigate the mechanism on Capparis spionosa L polysaccharide(CSPS) inducing apoptosis in HepG2 human hepatoma cell. MTT was ddopted to determine if CSPS had cytotoxic effect on HepG2. Morphology of HepG2 changed with dosages of CSPS was detected by laser confocal scanning microscope. Flow cytometry(FCM) was used to detect the apoptosis by PI labeling method. Calcium, mitochondrial membrane potential, Bcl-2/Bax of HepG2 cells were detected by laser confocal scanning microscope. The result of MTT showed that CSPS could inhibit the growth of HepG2 significantly. HepG2 cells were shrinkage, fragmentation, appearance of apoptotic bodies by laser confocal scanning microscope. HepG2 cell apoptosis rate was increased gradually with dosage by FCM. Calcium concentration, mitochondrial membrane potential, Bcl-2 protein of HepG2 were decreased, Bax protein content of HepG2 was increased by laser confocal scanning microscope. CSPS induced HepG2 apoptosis by controlling Bax/Bcl-2 in Ca2+ path.

scholarly journals Virulent Shigella flexneri Causes Damage to Mitochondria and Triggers Necrosis in Infected Human Monocyte-Derived Macrophages

2005 ◽  
Vol 73 (1) ◽  
pp. 504-513 ◽  
Author(s):  
James F. Koterski ◽  
Massoumeh Nahvi ◽  
Malabi M. Venkatesan ◽  
Beatrice Haimovich
Keyword(s):  
Membrane Potential ◽  
Cell Membrane ◽  
Membrane Permeability ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Shigella Flexneri ◽  
Human Monocyte ◽  
Cell Membrane Permeability ◽  
Vital Dyes

ABSTRACT Shigella flexneri is a gram-negative bacterium that causes bacillary dysentery in humans that is characterized by an acute inflammatory response of the colon. The fate of phagocytes that are infected in vitro with virulent Shigella has been the subject of some investigation and debate. In this study we found that virulent Shigella caused a rapid increase in the cell membrane permeability of infected human monocyte-derived macrophages (HMDM) but not in the cell membrane permeability of monocytes, as demonstrated by the uptake of fluorescent vital dyes. Within 2 h of infection, 59% ± 6% of the HMDM and ≤4% of the monocytes were stained with propidium iodide. Treatment of the cells with the inhibitors of caspases YVAD and zVAD, the antioxidants N-acetyl-l-cysteine and butylated hydroxyanisole, or an inhibitor of NADPH oxidase, diphenyleniodonium, did not alter the infection outcome. Importantly, we found that virulent Shigella caused a rapid drop in the ATP level to about 50% in infected HMDM. Furthermore, using a combination of fluorescent vital dyes and mitochondrial membrane potential-sensitive dyes, we observed that cells that exhibited a permeable cell membrane were not stained by the mitochondrion-specific dyes, indicating that the mitochondrial membrane potential was lost in these cells. We also observed infected cells that were not stained with either type of dye, indicating that the loss of the mitochondrial membrane potential preceded the increase in cell membrane permeability. Taken together, our studies showed that virulent Shigella flexneri targets the host cell mitochondria for destruction. This activity may account for the necrotic cell death precipitated by these pathogens.

scholarly journals Antiproliferative activity of Syzygium coriaceum, an endemic plant of Mauritius, with its UPLC-MS metabolite fingerprint: A mechanistic study

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252276
Author(s):  
Nawraj Rummun ◽  
Ahmed Serag ◽  
Philippe Rondeau ◽  
Srishti Ramsaha ◽  
Emmanuel Bourdon ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Untreated Control ◽  
Metabolite Profiling ◽  
Mitochondrial Membrane ◽  
Leaf Extract ◽  
Biological Activities ◽  
Mechanistic Study ◽  
Endemic Plant

Flowering plants from the Syzygium genus have long been used in different ethnomedicinal systems worldwide and have been under scrutiny for their biological activities. Syzygium coriaceum, an endemic plant of Mauritius has been poorly studied for its potential application against cancer. Herein, Syzygium coriaceum leaf extract has been investigated for its anticancer effect against hepatocellular carcinoma (HepG2) cells. The anticancer activity was assessed using cell proliferation assays, flow cytometry, JC-1 mitochondrial membrane potential assay, and the COMET assay. Un-targeted metabolite profiling via ultra-performance liquid chromatography coupled to high-resolution qTOF-MS (UPLC-MS) and aided by molecular networking was employed to identify the crude extract metabolites. S. coriaceum treatment induced a dose-dependent increase in lactate dehydrogenase leakage into the culture media, peaking up to 47% (p ≤ 0.0001), compared to untreated control. Moreover, at 40 μg/mL, S. coriaceum led to 88.1% (p ≤ 0.0001) drop in mitochondrial membrane potential and 5.7% (p ≤ 0.001) increased in the number of the cell population in G0/G1 phase as well as increased (p < 0.05) the proportion of cells undergoing apoptotic/necrotic cell death. More so, at 10 μg/mL, S. coriaceum induced DNA damage which was 19 folds (p < 0.001) higher than that of untreated control cells. Metabolite profiling indicated the presence of 65 metabolites, out of which 59 were identified. Tannins, flavonoids, nitrogenous compounds, and organic acids were the most predominant classes of compounds detected. Our findings showed that the presence of tannins and flavonoids in S. coriaceum leaf extract could account for the multiple mechanisms of actions underlying the antiproliferative effect against HepG2 cells.

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