scholarly journals Non-esterified Fatty Acid-Induced Reactive Oxygen Species Mediated Granulosa Cells Apoptosis Is Regulated by Nrf2/p53 Signaling Pathway

Antioxidants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 523
Author(s):  
Yiru Wang ◽  
Chengmin Li ◽  
Julang Li ◽  
Genlin Wang ◽  
Lian Li
Keyword(s):  
Reactive Oxygen Species ◽  
Fatty Acid ◽  
Granulosa Cells ◽  
Ovarian Function ◽  
Reactive Oxygen ◽  
Follicular Development ◽  
Oxygen Species ◽  
Related Protein ◽  
Induced Apoptosis

Negative energy balance (NEB) during the perinatal period can affect dairy cow follicular development and reduce the fecundity. Non-esterified fatty acid (NEFA) concentration is elevated during NEB, and is known to be toxic for multiple cell types. In the ovary, NEB increased NEFA, and may influences follicular growth and development. However, the effect and mechanism of NEFA on granulosa cells (GCs) in vitro remains unknown. In this study, we found that NEFA dose-dependently induced apoptosis in primary cultured granulosa cells. Mechanistically, our data showed that NEFA significantly increased reactive oxygen species (ROS) levels, resulting in the activation of endoplasmic reticulum stress (ERS) and eventually cell apoptosis in GCs. Moreover, NEFA also increased the phosphorylation levels of ERK1/2 and p38MAPK pathways, upregulated the expression of p53 and potentially promoted its translocation to the nuclear, thus transcriptionally activated Bax, a downstream gene of this pathway. NEFA also promoted nuclear factor E2 (Nrf2) expression and its level in the nuclear. To elucidate the mechanism of NEFA action, N-acetyl-l-cysteine (NAC), a ROS scavenger was used to verify the role of ROS in NEFA induced apoptosis of GCs. NAC pretreatment reversed the NEFA-induced ERS-related protein and apoptosis-related protein levels. Meanwhile, NAC pretreatment also blocked the phosphorylation of ERK1/2 and p38 induced by NEFA, and the nucleation of Nrf2 and p53, suggesting that ROS plays a crucial role in regulating the NEFA-induced apoptosis of GCs. Together, these findings provide an improved understanding of the mechanisms underlying GCs apoptosis, which could potentially be useful for improving ovarian function.

scholarly journals In vitro Studies on Calcium Oxalate Induced Apoptosis Attenuated by Didymocarpus pedicellata

2021 ◽  
Vol 12 (6) ◽  
pp. 7342-7355
Keyword(s):  
Oxidative Stress ◽  
Reactive Oxygen Species ◽  
Cell Death ◽  
Calcium Oxalate ◽  
Reactive Oxygen ◽  
Ethanolic Extract ◽  
Sem Analysis ◽  
Oxygen Species ◽  
Induced Apoptosis

The present study focuses on exploring the antilithiatic potential of Didymocarpus pedicellata, which is valuable in managing renal disorders. Urolithiasis is an idiopathic disorder with a high recurrence and an incidence rate and is of major concern worldwide due to partial and unsatisfactory relief. Calcium oxalate crystals in contact with renal epithelial cells (HK2), causing reactive oxygen species overproduction, oxidative stress, apoptosis resulting in crystal adhesion and internalization. Crystals were modulated by cotreatment with ethanolic extract of D. pedicellata. Cell toxicity assay was assessed using flow cytometry. Cell-crystal interaction, adhesion, and internalization were visualized through Scanning electron microscopy (SEM) analysis and hematoxylin-eosin staining. The lithogenic induction caused impairment of renal function due to oxidative stress, measured by ROS levels. Cell death assays were detected by dual staining methods. Fluorimeter evaluation pointed to active caspase 3 mediated cell death (apoptotic) in oxalate injured cells was attenuated by Didymocarpus pedicellata extract. Alterations in cell adhesion were observed by immunocytochemistry. The current study revealed that the Didymocarpus pedicellata was endowed with antiurolithiatic activity as it displayed increased viability, reduced oxidative stress due to lowered production of intracellular reactive oxygen species (ROS), and decreased apoptosis when oxalate injured HK2 cells were cotreated with the extract.

scholarly journals Arachidonic acid-induced apoptosis in rat hepatoma AS-30D cells is mediated by reactive oxygen species.

2009 ◽  
Vol 56 (4) ◽  
Author(s):  
Dorota Dymkowska ◽  
Lech Wojtczak
Keyword(s):  
Reactive Oxygen Species ◽  
Arachidonic Acid ◽  
Apoptotic Cell ◽  
Reactive Oxygen ◽  
Water Soluble ◽  
Oxygen Species ◽  
Irreversible Inhibitor ◽  
Induced Apoptosis ◽  
Rat Hepatoma

Arachidonic acid at micromolar concentrations produced a drastic increase of the generation of reactive oxygen species (ROS) in rat hepatoma AS-30D cells cultivated in vitro along with an increase in the incidence of apoptotic cell death. Both processes were prevented by trolox, a water-soluble tocopherol derivative, and tempol, a known antioxidative agent. A synthetic hybrid of lipoic acid and trolox or preincubation with N-acetylcysteine were less effective. Preincubation of the cells with etomoxir, a known highly specific irreversible inhibitor of carnitine-palmitoyltransferase I, partly decreased the ROS formation induced by arachidonic acid but it did not affect the increase in apoptosis. Cumulatively, these results indicate that apoptosis induced in hepatoma cells by arachidonic acid is mediated by ROS. They also suggest that this effect is due to arachidonic acid as such and not to its mitochondrial oxidative metabolites.

Antifilarial activity of azadirachtin fuelled through reactive oxygen species induced apoptosis: a thorough molecular study on Setaria cervi

2018 ◽  
Vol 93 (05) ◽  
pp. 519-528 ◽  
Author(s):  
N. Mukherjee ◽  
N. Joardar ◽  
S.P. Sinha Babu
Keyword(s):  
Reactive Oxygen Species ◽  
Lethal Dose ◽  
Reactive Oxygen ◽  
Molecular Study ◽  
Oxygen Species ◽  
Gene Expressions ◽  
Setaria Cervi ◽  
Induced Apoptosis

AbstractEfficacious therapeutic strategies against lymphatic filariasis are always sought after. However, natural products are a promising resource for developing effective antifilarial agents. Azadirachtin, a significant tetranortriterpenoid phytocompound found in Azadirachta indica, was evaluated in vitro for antifilarial potential against the filarial parasite Setaria cervi. Dye exclusion and MTT assay confirmed the antifilarial potential of azadirachtin against S. cervi with a median lethal dose (LC50) of 6.28 μg/ml for microfilariae (mf), and 9.55 μg/ml for adult parasites. Morphological aberrations were prominent in the histological sections of the azadirachtin-exposed parasites. Moreover, alterations in the reactive oxygen species (ROS) parameters in treated parasites were evident. Induction of apoptosis in treated parasites was confirmed by DNA laddering, acridine orange (AO)/ethidium bromide (EtBr) double staining and in situ DNA fragmentation. The downregulation of anti-apoptotic CED-9 and upregulation of proapoptotic EGL-1, CED-4 and CED-3 at both the transcription and translation levels confirmed apoptosis execution at the molecular level. Changes in the gene expressions of nuc-1, cps-6 and crn-1 further clarified the molecular cause of DNA degradation. Furthermore, azadirachtin was found to be non-toxic in both in vitro and in vivo toxicity analyses. Therefore, the experimental evidence detailed the pharmacological effectiveness of azadirachtin as a possible therapeutic agent against filariasis.

scholarly journals Follicle stimulating hormone inhibits the expression of p53 up-regulated modulator of apoptosis induced by reactive oxygen species through PI3K/Akt in mouse granulosa cells

2020 ◽  
pp. 687-694
Author(s):  
XY Shi ◽  
ZQ Guan ◽  
JN Yu ◽  
HL Liu
Keyword(s):  
Reactive Oxygen Species ◽  
Mrna Level ◽  
Follicle Stimulating Hormone ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Follicular Atresia ◽  
Puma Expression ◽  
Induced Apoptosis

In mammalian ovaries, follicular atresia occurs periodically and destroys almost all the follicles in the ovary. Follicle-stimulating hormone (FSH) acts as the primary survival factor during follicular atresia by preventing apoptosis in granulosa cells (GCs). Many studies have demonstrated that oxidative stress-induced apoptosis is a main cause of follicular atresia. Reactive oxygen species (ROS)-induced GCs apoptosis is regulated by a variety of signaling pathways involving numerous genes and transcription factors. Therefore, we examined whether FSH inhibits the expression of p53 up-regulated modulator of apoptosis (PUMA) induced by reactive oxygen species (ROS) through phosphoinositide 3-kinase (PI3K) / protein kinase B (AKT) in mouse GCs. In vivo study: thirty-two-mice were randomly assigned to four groups and given FSH. We found that FSH can inhibit the 3-nitropropionic acid (3-NP) induced apoptosis and PUMA expression in mRNA level. Moreover, In vitro experiment, we found that FSH can inhibit the H(2)O(2)-induced apoptosis and PUMA expression in mRNA level. Additionally, we also found that PI3K/AKT inhibitor LY294002 abolished the downregulation of PUMA mRNA by FSH in vitro, In conclusion, FSH inhibit the expression of PUMA induced by ROS through PI3K/AKT pathway in vivo and vitro.

scholarly journals rno-miR-128-3p promotes apoptosis in rat granulosa cells (GCs) induced by norepinephrine through Wilms tumor 1 (WT1)

2021 ◽  
Author(s):  
Ming Li ◽  
Ling Xue ◽  
Weibin Xu ◽  
Pingping Liu ◽  
Feng Li
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Ovarian Function ◽  
Wilms Tumor ◽  
Sympathetic Innervation ◽  
Follicular Development ◽  
Wilms Tumor 1 ◽  
Ovarian Follicular Development ◽  
Induced Apoptosis

AbstractThe mechanism related to ovarian follicular is complex, which has not been fully elucidated. Abundant reports have confirmed that the ovarian function development is closely related to sympathetic innervation. As one of the major neurotransmitters, norepinephrine (NE) is considered an effective regulator of ovarian functions like granulosa cell (GC) apoptosis. However, the mechanism between NE and GC apoptosis in rat is still unclear. In our study, GCs were isolated and cultured in vitro with NE treatment. The apoptosis of GCs was facilitated by NE. Wilms tumor 1 (WT1) was found to be significantly downregulated in GCs after NE treatment, and overexpression of WT1 repressed apoptosis in rat GCs induced by NE. rno-miR-128-3p was found to be significantly enhanced by NE in GCs, and inhibition of rno-miR-128-3p repressed apoptosis in rat GCs induced by NE. Mechanistically, rno-miR-128-3p interacted with WT1 and repressed its expression. In summary, inhibition of rno-miR-128-3p may enhance WT1 expression, and then repress NE-induced apoptosis in rat GCs. Our research may provide a new insight for the improvement of ovarian follicular development.

scholarly journals Norepinephrine, Active Norepinephrine Transporter, and Norepinephrine-Metabolism Are Involved in the Generation of Reactive Oxygen Species in Human Ovarian Granulosa Cells

Endocrinology ◽  
2012 ◽  
Vol 153 (3) ◽  
pp. 1472-1483 ◽  
Author(s):  
S. Saller ◽  
J. Merz-Lange ◽  
S. Raffael ◽  
S. Hecht ◽  
R. Pavlik ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Monoamine Oxidase ◽  
Cellular Uptake ◽  
Granulosa Cells ◽  
Reactive Oxygen ◽  
Monoamine Oxidase A ◽  
Oxygen Species ◽  
Uptake And Metabolism

The neurotransmitter norepinephrine (NE) is derived from the sympathetic nervous system and may be involved in the regulation of ovarian functions. Ovarian innervation increases in patients with polycystic ovarian syndrome (PCOS), prompting us to readdress a role of NE in the human ovary. In vitro fertilization-derived granulosa cells (GC), follicular fluids (FF), and ovarian sections were studied. NE was found in FF and freshly isolated GC, yet significantly lower levels of NE were detected in samples from PCOS patients. Furthermore, the metabolite normetanephrine was detected in FF. Together this suggests cellular uptake and metabolism of NE in GC. In accordance, the NE transporter and NE-metabolizing enzymes [catechol-o-methyltransferase (COMT) and monoamine oxidase A] were found in GC, COMT in GC and thecal cells of large human antral follicles in vivo and in cultured GC. Cellular uptake and metabolism of NE also occurred in cultured GC, events that could be blocked pharmacologically. NE, in the range present in FF, is unlikely to affect GC via activation of typical α- or β-receptors. In line with this assumption, it did not alter phosphorylation of MAPK. However, NE robustly induced the generation of reactive oxygen species (ROS). This action occurred even when receptors were blocked but was prevented by blockers of NE transporter, COMT, and monoamine oxidase A. Thus, NE contributes to the microenvironment of preovulatory human follicles and is lower in PCOS. By inducing the production of ROS in GC, NE is linked to ROS-regulated events, which are emerging as crucial factors in ovarian physiology, including ovulation.

scholarly journals A nitric oxide-releasing prodrug promotes apoptosis in human renal carcinoma cells: Involvement of reactive oxygen species

2021 ◽  
Vol 19 (1) ◽  
pp. 635-645
Author(s):  
Jindong Xie ◽  
Lieqian Chen ◽  
Dongqiang Huang ◽  
Weiwei Yue ◽  
Jingyu Chen ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Reactive Oxygen Species ◽  
Renal Carcinoma ◽  
Reactive Oxygen ◽  
Carcinoma Cells ◽  
Oxygen Species ◽  
Induced Apoptosis ◽  
Human Renal Carcinoma ◽  
Renal Carcinoma Cells

Abstract Background JS-K is a nitric oxide (NO)-releasing prodrug of the O2-arylated diazeniumdiolate group that shows pronounced cytotoxicity and antitumor properties in numerous cancer models, including in vitro as well as in vivo. Reactive oxygen species (ROS) induce carcinogenesis by altering the redox status, causing increment in vulnerability to oxidative stress. Material and methods To determine the effect of JS-K, a glutathione S-transferase (GST)-activated NO-donor prodrug, on the induction of ROS accumulation, proliferation, and apoptosis in human renal carcinoma cells, we measured the changes of cell proliferation, apoptosis, ROS growth, and initiation of the mitochondrial signaling pathway before and after JS-K treatment. Results In vitro, dose- and time-dependent development of renal carcinoma cells were controlled for JS-K, and JS-K also triggered ROS aggregation and cell apoptosis. Treatment with JS-K induces the levels of pro-apoptotic proteins (Bak and Bax) and decrease the number of anti-apoptotic protein (Bcl-2). In fact, JS-K-induced apoptosis was reversed by the antioxidant N-acetylcysteine, and oxidized glutathione, a pro-oxidant, improved JS-K-induced apoptosis. Finally, we demonstrated that in renal carcinoma cells, JS-K improved the chemosensitivity of doxorubicin. Conclusion Our data indicate that JS-K-released NO induce apoptosis of renal carcinoma cells by increasing ROS levels.

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