Madecassic acid protects human periodontal ligament fibroblasts against hydrogen peroxide-induced cell damage by maintaining mitochondrial membrane potential

Hydrogen peroxide (H2O2) and hypochlorite (HOCl) cause a variety of cellular dysfunctions. In this study we examined the effects of these agents on the electrical potential gradient across the inner membrane of mitochondria in situ in isolated rat heart myocytes. Myocytes were prepared by collagenase digestion and incubated in the presence of H2O2 or HOCl. Transmembrane electrical gradients were measured by distribution of [3H]triphenylmethylphosphonium+, a lipophilic cation. The particulate fraction was separated from the cytosolic compartment first by permeabilization using digitonin, followed by rapid centrifugal sedimentation through a bromododecane layer. We found that the mitochondrial membrane potential (161 ± 7 mV, negative inside) was relatively well maintained under oxidant stress, i.e., the potential was decreased only at high concentrations of HOCl and H2O2 and gradually with time. The membrane potential of isolated rat heart mitochondria was affected similarly by H2O2 and HOCl in a concentration- and time-dependent manner. High concentrations of oxidants also reduced the cellular ATP level but did not significantly change the matrix volume. When the extra-mitochondrial free calcium concentration was increased in permeabilized myocytes, the transmembrane potential was decreased proportionally, and this decrease was potentiated further by H2O2. These results support the view that heart mitochondria are equipped with well-developed defense mechanisms against oxidants, but the action of H2O2 on the transmembrane electrical gradient is exacerbated by an increase in cytosolic calcium. Keywords: ATP, calcium, cardiomyocyte, cell defense, mitochondrial membrane potential, oxidant, triphenylmethylphosphonium.

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Seminal plasma has limited counteracting effects following induction of oxidative stress in donkey spermatozoa

Reproduction Fertility and Development ◽

10.1071/rd19192 ◽

2020 ◽

Vol 32 (6) ◽

pp. 619

Author(s):

Marion Papas ◽

Jaime Catalan ◽

Sebastián Bonilla-Correal ◽

Sabrina Gacem ◽

Jordi Miró ◽

...

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Sperm Motility ◽

Seminal Plasma ◽

Mitochondrial Membrane ◽

Skim Milk ◽

Sperm Parameters ◽

High Concentration

The aim of this study was to evaluate the response of donkey spermatozoa to oxidative stress induced by hydrogen peroxide, and to determine whether the presence of seminal plasma modulates the sperm response to that stress. Nine ejaculates were collected, extended in skim milk extender and split into two aliquots. Seminal plasma was removed from the first but not second aliquot. Samples were subsequently split into four aliquots supplemented with different concentrations of commercial hydrogen peroxide (0, 100 and 250µM and 50mM). Aliquots were incubated at 37°C under aerobic conditions and several sperm parameters, namely motility, viability, intracellular levels of peroxides and superoxides and mitochondrial membrane potential, were evaluated at 0, 1 and 3h. Exposure to hydrogen peroxide markedly decreased sperm motility but had much less of an effect on sperm viability, mitochondrial membrane potential and intracellular reactive oxygen species levels. A protective effect of seminal plasma against the loss of sperm motility was not apparent, but some kinetic parameters and relative levels of superoxides were better maintained when seminal plasma was present together with high concentration of hydrogen peroxide. In conclusion, oxidative stress induced by hydrogen peroxide reduces donkey sperm motility and has a less apparent effect on other sperm parameters. Finally, seminal plasma is only able to partially ameliorate the detrimental effect of this induced stress.

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Involvement of Hydrogen Peroxide in the Manganese-Induced Myocytes Mitochondrial Membrane Potential Loss

Toxicology Mechanisms and Methods ◽

10.1080/15376510802428583 ◽

2008 ◽

Vol 19 (1) ◽

pp. 66-72 ◽

Cited By ~ 2

Author(s):

Huijuan Yang ◽

Ling Gu ◽

Le Zhang ◽

Ming Yan ◽

Xiaoxiang Zheng

Keyword(s):

Hydrogen Peroxide ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Potential Loss ◽

Mitochondrial Membrane Potential Loss

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Rise in cytosolic Ca2+ and collapse of mitochondrial potential in anoxic, but not hypoxic, rat proximal tubules.

Journal of the American Society of Nephrology ◽

10.1681/asn.v7112348 ◽

1996 ◽

Vol 7 (11) ◽

pp. 2348-2356

Author(s):

S M Peters ◽

M J Tijsen ◽

R J Bindels ◽

C H Van Os ◽

J F Wetzels

Keyword(s):

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Cell Injury ◽

Cell Damage ◽

Energy State ◽

Oxygen Deprivation ◽

Proximal Tubules ◽

Rhodamine 123 ◽

Anoxic Conditions

It has been suggested that ischemic renal proximal tubular cell injury is mediated by an increase in cytosolic calcium concentrations ((Ca2+)i). However, measurements of (Ca2+)i in rat or rabbit proximal tubules exposed to hypoxia or anoxia have yielded ambiguous results. This study explored the possibility that the severity of oxygen deprivation and the energy state of the mitochondria are important determinants of (Ca2+)i. To this end, (Ca2+)i (measured with fura-2) and the mitochondrial membrane potential (measured with rhodamine 123) were studied simultaneously in individual rat proximal tubules in hypoxic and anoxic conditions. (Ca2+)i did not change during hypoxia, but increased rapidly during anoxia. Increases in (Ca2+)i were only observed in parallel with a decrease of rhodamine 123 fluorescence, which indicates a collapse of the mitochondrial membrane potential. The increase in (Ca2+)i during anoxia was prevented by incubating the tubules in a low Ca2+ medium, which did not interfere with the collapse of the mitochondrial membrane potential. Both hypoxic and anoxic incubation led to cell death, as assessed by the fluorescent dye propidium iodide. These results clearly demonstrate that the level of oxygen deprivation is critical in determining changes in (Ca2+)i. Because cell damage occurred in both hypoxic and anoxic conditions. It was concluded that an increase in (Ca2+)i is not a necessary prerequisite for the development of ischemic cell injury.

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CoCl2 induces apoptosis via a ROS-dependent pathway and Drp1-mediated mitochondria fission in periodontal ligament stem cells

AJP Cell Physiology ◽

10.1152/ajpcell.00248.2017 ◽

2018 ◽

Vol 315 (3) ◽

pp. C389-C397 ◽

Cited By ~ 14

Author(s):

Yuting He ◽

Xueqi Gan ◽

Ling Zhang ◽

Beilei Liu ◽

Zhuoli Zhu ◽

...

Keyword(s):

Stem Cells ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Periodontal Ligament ◽

Mitochondrial Membrane ◽

Mitochondrial Fission ◽

Orthodontic Appliances ◽

Dependent Manner ◽

Periodontal Ligament Stem Cells ◽

Atp Level

Oxygen deficiency is associated with various oral diseases, including chronic periodontitis, age-related alveolar bone loss, and mechanical stress-linked cell injury from orthodontic appliances. Nevertheless, our understanding of the impact of hypoxia on periodontal tissues and its biochemical mechanism is still rudimentary. The purpose of this research was to elucidate the effects of hypoxia on the apoptosis of human periodontal ligament stem cells (PDLSCs) in vitro and the underlying mechanism. Herein, we showed that cobalt chloride (CoCl2) triggered cell dysfunction in human PDLSCs in a concentration-dependent manner and resulted in cell apoptosis and oxidative stress overproduction and accumulation in PDLSCs. In addition, CoCl2 promoted mitochondrial fission in PDLSCs. Importantly, CoCl2 increased the expression of dynamin-related protein 1 (Drp1), the major regulator in mitochondrial fission, in PDLSCs. Mitochondrial division inhibitor-1, pharmacological inhibition of Drp1, not only inhibited mitochondrial fission but also protected against CoCl2-induced PDLSC dysfunction, as shown by increased mitochondrial membrane potential, increased ATP level, reduced reactive oxygen species (ROS) level, and decreased apoptosis. Furthermore, N-acety-l-cysteine, a pharmacological inhibitor of ROS, also abolished CoCl2-induced expression of Drp1 and protected against CoCl2-induced PDLSC dysfunction, as shown by restored mitochondrial membrane potential, ATP level, inhibited mitochondrial fission, and decreased apoptosis. Collectively, our data provide new insights into the role of the ROS-Drp1-dependent mitochondrial pathway in CoCl2-induced apoptosis in PDLSCs, indicating that ROS and Drp1 are promising therapeutic targets for the treatment of CoCl2-induced PDLSC dysfunction.

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Effect of Single Layer Centrifugation on reactive oxygen species and sperm mitochondrial membrane potential in cooled stallion semen

Reproduction Fertility and Development ◽

10.1071/rd15440 ◽

2017 ◽

Vol 29 (5) ◽

pp. 1039 ◽

Cited By ~ 16

Author(s):

J. M. Morrell ◽

A. Lagerqvist ◽

P. Humblot ◽

A. Johannisson

Keyword(s):

Reactive Oxygen Species ◽

Hydrogen Peroxide ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Reactive Oxygen ◽

Single Layer ◽

Ros Production ◽

Oxygen Species ◽

Species Production

Additional means are needed for evaluating the quality of stallion spermatozoa in semen doses for AI. Mitochondrial membrane potential (ΔΨm) has been linked to fertility in some species, but is rarely used in the evaluation of cooled stallion semen; metabolic activity may be associated with reactive oxygen species production (ROS). In the present study, ΔΨm and ROS production were measured in doses of cooled stallion semen. The effect of colloid centrifugation on these parameters was also investigated. In this case, colloid centrifugation involves centrifuging a sperm sample through a silane-coated silica colloid formulation to retrieve the most robust spermatozoa. High and low ΔΨm in cooled stallion semen varied between stallions and between ejaculates, but was not affected by single-layer centrifugation (SLC). The SLC-selected spermatozoa produced significantly less hydrogen peroxide than controls (P < 0.001), which could explain the increased longevity and retention of fertilising capacity seen in previous studies. For SLC samples, ΔΨm was positively associated with viable spermatozoa that were not producing reactive oxygen species (r = 0.49; P < 0.001) and negatively associated with ROS production (for superoxide: r = –0.4, P < 0.01; for hydrogen peroxide: r = –0.39, P < 0.05). There was no clear association between ΔΨm and ROS production in control samples.

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Flurochloridone Induced Abnormal Spermatogenesis by Damaging Testicular Sertoli Cells in Mice

10.21203/rs.3.rs-1151621/v1 ◽

2021 ◽

Author(s):

Weiqi Sun ◽

Fang Tian ◽

Hongjie Pan ◽

Xiuli Chang ◽

Minjie Xia ◽

...

Keyword(s):

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Sertoli Cell ◽

Sertoli Cells ◽

Mitochondrial Membrane ◽

Cell Damage ◽

Sperm Quality ◽

Reproductive Toxicity ◽

Sperm Concentration ◽

Mitochondrial Damage

Abstract BackgroundFlurochloridone (FLC), a selective herbicide used on a global scale, has been reported to have male reproductive toxicity which evidence is limited and the mechanism is still unclear. The present study was conducted to systematically explore the male reproductive toxicity of FLC, including sperm quality, spermatogenesis process, toxicity targets and possible mechanisms. MethodsMale C57BL/6 mice aged 6-7 weeks received gavage administration of FLC (365/730 mg/kg body weight) for 28 consecutive days. Then the tissue and sperm of mice were collected for analysis. We measured the coefficient of male reproductive organs, and analyzed sperm concentration, motility, malformation rate and mitochondrial membrane potential. Spermatocyte immunofluorescence staining was performed to analyze meiosis processes. At the same time, we performed pathological staining on the testis and epididymis tissue, and performed TUNEL staining, immunohistochemical analysis and ultrastructural observation on the testicular tissue.ResultsThe results showed that FLC caused mice testicular weight reduction, dysfunction and architectural damage, but no significant adverse effect was found in epididymis. The exposure interfered with the proliferation of spermatogonia and the process of meiosis, affecting sperm concentration, motility, kinematic parameters, morphology and mitochondrial membrane potential, leading to sperm quality decline. Furthermore, mitochondrial damage and apoptosis of testicular Sertoli cells were observed in mice treated with FLC. ConclusionWe found that FLC has significant adverse effects on spermatogonia proliferation and meiosis. Meanwhile, apoptosis and mitochondrial damage may be the potential mechanism of Sertoli cell damage. Our study demonstrated that FLC could induce testicular Sertoli cell damage, leading to abnormal spermatogenesis which resulted in sperm quality decline and provided a methodological reference for related studies.

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Protective Effect of Ginsenoside Rg1 on Oxidative Damage Induced by Hydrogen Peroxide in Chicken Splenic Lymphocytes

Oxidative Medicine and Cellular Longevity ◽

10.1155/2019/8465030 ◽

2019 ◽

Vol 2019 ◽

pp. 1-13 ◽

Cited By ~ 5

Author(s):

Shicheng Bi ◽

Xiaodan Ma ◽

Yuemin Wang ◽

Xiaoqing Chi ◽

Yong Zhang ◽

...

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Membrane Potential ◽

Cell Viability ◽

Mitochondrial Membrane Potential ◽

Transcriptome Analysis ◽

Mitochondrial Membrane ◽

Receptor Interaction ◽

Apoptotic Cells ◽

Ginsenoside Rg1

Previous investigation showed that ginsenoside Rg1 (Rg1) extracted from Panax ginseng C.A. Mey has antioxidative effect on oxidative stress in chickens. The present study was designed to investigate the protective effects of Rg1 on chicken lymphocytes against hydrogen peroxide-induced oxidative stress and the potential mechanisms. Cell viability, apoptotic cells, malondialdehyde, activity of superoxide dismutase, mitochondrial membrane potential, and [Ca2+]i concentration were measured, and transcriptome analysis and quantitative real-time polymerase chain reaction were used to investigate the effect of Rg1 on gene expression of the cells. The results showed that treatment of lymphocytes with H2O2 induced oxidative stress and apoptosis. However, pretreatment of the cells with Rg1 dramatically enhanced cell viability, reduced apoptotic cells, and decreased oxidative stress induced by H2O2. In addition, Rg1 reduced these H2O2-dependent decreases in mitochondrial membrane potential and reversed [Ca2+]i overload. Transcriptome analysis showed that 323 genes were downregulated and 105 genes were upregulated in Rg1-treated cells. The differentially expressed genes were involved in Toll-like receptors, peroxisome proliferator-activated receptor signaling pathway, and cytokine-cytokine receptor interaction. The present study indicated that Rg1 may act as an antioxidative agent to protect cell damage caused by oxidative stress via regulating expression of genes such as RELT, EDA2R, and TLR4.

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Gold Nanoparticles Induce Oxidative Stress and Apoptosis in Human Kidney Cells

Nanomaterials ◽

10.3390/nano10050995 ◽

2020 ◽

Vol 10 (5) ◽

pp. 995 ◽

Cited By ~ 2

Author(s):

Maria Enea ◽

Eulália Pereira ◽

Miguel Peixoto de Almeida ◽

Ana Margarida Araújo ◽

Maria de Lourdes Bastos ◽

...

Keyword(s):

Gold Nanoparticles ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Cell Damage ◽

Human Kidney ◽

In Vivo Studies ◽

Dependent Manner ◽

Concentration Dependent Manner

Gold nanoparticles (AuNPs) are highly attractive for biomedical applications. Therefore, several in vitro and in vivo studies have addressed their safety evaluation. Nevertheless, there is a lack of knowledge regarding their potential detrimental effect on human kidney. To evaluate this effect, AuNPs with different sizes (13 nm and 60 nm), shapes (spheres and stars), and coated with 11-mercaptoundecanoic acid (MUA) or with sodium citrate, were synthesized, characterized, and their toxicological effects evaluated 24 h after incubation with a proximal tubular cell line derived from normal human kidney (HK-2). After exposure, viability was assessed by the MTT assay. Changes in lysosomal integrity, mitochondrial membrane potential (ΔΨm), reactive species (ROS/RNS), intracellular glutathione (total GSH), and ATP were also evaluated. Apoptosis was investigated through the evaluation of the activity of caspases 3, 8 and 9. Overall, the tested AuNPs targeted mainly the mitochondria in a concentration-dependent manner. The lysosomal integrity was also affected but to a lower extent. The smaller 13 nm nanospheres (both citrate- and MUA-coated) proved to be the most toxic among all types of AuNPs, increasing ROS production and decreasing mitochondrial membrane potential (p ≤ 0.01). For the MUA-coated 13 nm nanospheres, these effects were associated also to increased levels of total glutathione (p ≤ 0.01) and enhanced ATP production (p ≤ 0.05). Programmed cell death was detected through the activation of both extrinsic and intrinsic pathways (caspase 8 and 9) (p ≤ 0.05). We found that the larger 60 nm AuNPs, both nanospheres and nanostars, are apparently less toxic than their smaller counter parts. Considering the results herein presented, it should be taken into consideration that even if renal clearance of the AuNPs is desirable, since it would prevent accumulation and detrimental effects in other organs, a possible intracellular accumulation of AuNPs in kidneys can induce cell damage and later compromise kidney function.

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