ABSTRACT
The aim of the present study was to determine whether adipokines monocyte chemoattractant protein-1 (MCP-1) and plasminogen activator inhibitor-1 (PAI-1) can affect the functions of ovarian cells in cats. The addition of either MCP-1 or PAI-1 increased viability; promoted the accumulation of proliferation markers and progesterone and estradiol release; and decreased the accumulation of apoptosis markers in cultured feline granulosa cells. The present observations suggest that MCP-1 or PAI-1 can be physiological stimulators of ovarian granulosa cell functions.
ABSTRACT
Autophagy, an intrinsic catabolic mechanism that eliminates misfolded proteins, dysfunctional organelles, and lipid droplets, plays a vital function in energy balance and cytoplasmic quality control, in addition to maintaining cellular homeostasis. Liver cancer such as hepatocellular carcinoma (HCC) is one of the most common causes of cancer deaths globally and shows resistance to several anticancer drugs. Despite the rising incidence and poor prognosis of malignant HCC, the underlying molecular mechanisms driving this aggressive cancer remain unclear. Several natural compounds, such as phytochemicals of dietary and non-dietary origin, affect hepatocarcinogenesis signaling pathways in vitro and in vivo, which may help prevent and treat HCC cells. Current HCC cells treatments include chemotherapy, radiation, and surgery. However, these standard therapies have substantial side effects, and combination therapy enhances side effects for an acceptable therapeutic benefit. Therefore, there is a need to develop treatment strategies for HCC cells that are more efficacious and have fewer adverse effects. Multiple genetic and epigenetic factors are responsible for the HCC cells to become resistant to standard treatment. Autophagy contributes to maintain cellular homeostasis, which activates autophagy for biosynthesis and mitochondrial regulation and recycling. Therefore, modifying autophagic signaling would present a promising opportunity to identify novel therapies to treat HCC cells resistant to current standard treatments. This comprehensive review illustrates how natural compounds demonstrate their anti-hepatocellular carcinoma function through autophagy.
Subject(s)
Autophagy , Biological Products , Liver Neoplasms , Humans , Autophagy/drug effects , Liver Neoplasms/drug therapy , Liver Neoplasms/pathology , Liver Neoplasms/metabolism , Liver Neoplasms/genetics , Biological Products/pharmacology , Biological Products/therapeutic use , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/pathology , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/genetics , Animals , Signal Transduction/drug effectsABSTRACT
The aim of the present study was to determine the effects of the adipokines progranulin and omentin on the basic functions of feline ovarian cells. For this purpose, we investigated the effects of the addition of progranulin and omentin (0, 0.1, 1, or 10 ng/ml) on the proliferation (accumulation of PCNA and cyclin B1), apoptosis (accumulation of Bax and caspase 3) and progesterone release of cultured feline ovarian granulosa cells by quantitative immunocytochemistry and enzyme-linked immunosorbent assays (ELISAs). Both progranulin and omentin increased cell proliferation and decreased apoptosis. Both progranulin and omentin promoted progesterone release. The present findings demonstrate that the adipokines progranulin and omentin can directly regulate basic feline ovarian cell functions.
Subject(s)
Apoptosis , Cell Proliferation , Granulosa Cells , Animals , Female , Cats , Granulosa Cells/metabolism , Granulosa Cells/drug effects , Apoptosis/drug effects , Cell Proliferation/drug effects , Progesterone/metabolism , Progesterone/pharmacology , Progranulins/metabolism , Cytokines/metabolism , Cells, Cultured , Lectins/metabolism , Lectins/pharmacologyABSTRACT
This study aimed to investigate the impact of the epidermal growth factor receptor ligands amphiregulin (AREG) and epiregulin (EREG) on the fundamental functions of feline ovarian granulosa cells. Granulosa cells isolated from feline ovaries were incubated with AREG and EREG (0, 0.1, 1 or 10 ng/mL). The effects of these growth factors on cell viability, proliferation (assessed through BrdU incorporation), nuclear apoptosis (evaluated through nuclear DNA fragmentation) and the release of progesterone and estradiol were determined using Cell Counting Kit-8 assays, BrdU analysis, TUNEL assays and ELISAs, respectively. Both AREG and EREG increased cell viability, proliferation and steroid hormone release and reduced apoptosis. The present findings suggest that these epidermal growth factor receptor ligands may serve as physiological stimulators of feline ovarian cell functions.
Subject(s)
Amphiregulin , Apoptosis , Epiregulin , Granulosa Cells , Animals , Cats , Female , Amphiregulin/metabolism , Amphiregulin/genetics , Cell Proliferation , Cell Survival , Cells, Cultured , Epiregulin/metabolism , Epiregulin/genetics , Estradiol/metabolism , Granulosa Cells/cytology , Granulosa Cells/metabolism , Progesterone/metabolismABSTRACT
Autophagy is a globally conserved cellular activity that plays a critical role in maintaining cellular homeostasis through the breakdown and recycling of cellular constituents. In recent years, there has been much emphasis given to its complex role in cancer stem cells (CSCs) and stem cell treatment. This study examines the molecular processes that support autophagy and how it is regulated in the context of CSCs and stem cell treatment. Although autophagy plays a dual role in the management of CSCs, affecting their removal as well as their maintenance, the intricate interaction between the several signaling channels that control cellular survival and death as part of the molecular mechanism of autophagy has not been well elucidated. Given that CSCs have a role in the development, progression, and resistance to treatment of tumors, it is imperative to comprehend their biological activities. CSCs are important for cancer biology because they also show a tissue regeneration model that helps with organoid regeneration. In other words, the manipulation of autophagy is a viable therapeutic approach in the treatment of cancer and stem cell therapy. Both synthetic and natural substances that target autophagy pathways have demonstrated promise in improving stem cell-based therapies and eliminating CSCs. Nevertheless, there are difficulties associated with the limitations of autophagy in CSC regulation, including resistance mechanisms and off-target effects. Thus, the regulation of autophagy offers a versatile strategy for focusing on CSCs and enhancing the results of stem cell therapy. Therefore, understanding the complex interactions between autophagy and CSC biology would be essential for creating therapeutic treatments that work in both regenerative medicine and cancer treatment.
Subject(s)
Autophagy , Neoplastic Stem Cells , Humans , Neoplastic Stem Cells/metabolism , Neoplastic Stem Cells/pathology , Neoplasms/pathology , Neoplasms/therapy , Neoplasms/metabolism , Animals , Signal Transduction , Cell- and Tissue-Based Therapy/methods , Stem Cell TransplantationABSTRACT
The current understanding of the role of circular RNAs (circRNAs) in regulating ovarian functions is inadequate. To assess the impact of ciR-00596 and ciR-00646 on the regulation of basic porcine ovarian granulosa cell functions, we conducted upregulation (utilizing overexpressing vectors) and downregulation (utilizing shRNA vectors) of these circRNAs. The relative expression of both circRNAs, cell viability and proliferation (accumulation of PCNA, cyclin B1, and XTT-positive cells), cytoplasmic (accumulation of bax and caspase-3) and nuclear (DNA fragmentation) apoptosis, and the release of progesterone, testosterone, estradiol, IGF-I, and oxytocin were evaluated. Transfection of cells with the ciR-00596 overexpression vector resulted in increases in cell viability and proliferation and the release of progesterone and IGF-I, while it decreased the cytoplasmic and nuclear apoptosis, testosterone, estradiol, and oxytocin output. CiR-00596 inhibition had the opposite effects. The overexpression of ciR-00646 decreased cell viability and proliferation, and the release of progesterone, IGF-I, and oxytocin, while increasing cytoplasmic and nuclear apoptosis and the output of testosterone and estradiol. Our findings are the first to show the stimulatory action of ciR-00596 and the inhibitory effect of ciR-00646 on ovarian cell functions, including the cell cycle, apoptosis, and secretory activity.
Subject(s)
Apoptosis , Down-Regulation , Granulosa Cells , RNA, Circular , Up-Regulation , Animals , Female , RNA, Circular/metabolism , RNA, Circular/genetics , Swine , Granulosa Cells/metabolism , Granulosa Cells/physiology , Cell Proliferation/physiology , Cell Survival/physiology , Ovary/metabolism , Progesterone/metabolism , Estradiol/metabolism , Gene Expression Regulation/physiologyABSTRACT
The aim of this study was to analyze the phytochemical profile of Acacia cyclops trunk bark ethyl acetate extract using LC-tandem mass spectrometry for the first time, along with evaluating its antioxidant and anti-tyrosinase properties. Consequently, we determined the total phenolic and flavonoid contents of the extract under investigation and identified and quantified 19 compounds, including phenolic acids and flavonoids. In addition to assessing their antioxidant potential against DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis-[3-ethylbenzothiazoline-6-sulfonic] acid) assays, in vitro and in silico studies were conducted to evaluate the tyrosinase inhibitory properties of the A. cyclops extract. The ethyl acetate trunk bark extract exhibited a substantial total phenolic content and demonstrated significant antioxidant activity in terms of free radical scavenging, as well as notable tyrosinase inhibitory action (half-maximal inhibitory concentration [IC50] = 14.08 ± 1.10 µg/mL). The substantial anti-tyrosinase activity of the examined extract was revealed through molecular docking analysis and druglikeness prediction of the main selected compounds. The findings suggest that A. cyclops extract holds promise as a potential treatment for skin hyperpigmentation disorders.
Subject(s)
Acacia , Antioxidants , Enzyme Inhibitors , Molecular Docking Simulation , Monophenol Monooxygenase , Plant Bark , Plant Extracts , Monophenol Monooxygenase/antagonists & inhibitors , Acacia/chemistry , Plant Bark/chemistry , Antioxidants/pharmacology , Antioxidants/chemistry , Antioxidants/analysis , Plant Extracts/chemistry , Plant Extracts/pharmacology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/analysis , Phytochemicals/chemistry , Phytochemicals/pharmacology , Phytochemicals/analysis , Tandem Mass Spectrometry/methods , Flavonoids/chemistry , Flavonoids/analysis , Flavonoids/pharmacology , Phenols/chemistry , Phenols/analysis , Phenols/pharmacology , Chromatography, Liquid/methodsABSTRACT
The aim of the present study was to examine the influence of monocyte chemoattractant protein-1 (MCP-1) and plasminogen activator inhibitor-1 (PAI-1) on ovarian cell functions. Rabbit ovarian granulosa cells were cultured with or without MCP-1 or PAI-1 (at 0, 0.1, 1, or 10 ng/ml). Cell viability, proliferation, cytoplasmic apoptosis and release of progesterone and estradiol were measured by Cell Counting Kit-8 (CCK-8), BrdU incorporation, and cell death detection assays and ELISA. The addition of either MCP-1 or PAI-1 increased cell viability and proliferation and decreased apoptosis. MCP-1 promoted, while PAI-1 suppressed, progesterone release. Both MCP-1 and PAI-1 reduced estradiol output. The present results suggest that MCP-1 or PAI-1 can be physiological promoters of rabbit ovarian cell viability and proliferation, inhibitors of apoptosis and regulators of ovarian steroidogenesis.
Subject(s)
Apoptosis , Chemokine CCL2 , Granulosa Cells , Plasminogen Activator Inhibitor 1 , Progesterone , Animals , Female , Rabbits , Plasminogen Activator Inhibitor 1/genetics , Plasminogen Activator Inhibitor 1/metabolism , Granulosa Cells/drug effects , Granulosa Cells/physiology , Chemokine CCL2/genetics , Chemokine CCL2/metabolism , Apoptosis/drug effects , Progesterone/pharmacology , Estradiol/pharmacology , Cell Survival/drug effects , Cell Proliferation/drug effects , Cells, CulturedABSTRACT
Epigenetic methods to prevent the reproductive toxicity of oil-related environmental contaminants are currently unavailable. The present study aimed to examine the ability of the microRNA miR-152 to mitigate the effects of benzene on ovarian cells. Porcine ovarian granulosa cells transfected or not transfected with miR-152 mimics were cultured with or without benzene (0, 10 and 100 ng/ml). The expression of miR-152; viability; proliferation (cell proliferation and expression of mRNAs and accumulation of PCNA and cyclin B1); apoptosis (expression of mRNAs and accumulation of bax and caspase 3; and the proportion of cells with fragmented DNA); and release of progesterone, estradiol and IGF-I were analyzed via RT-qPCR; the Trypan blue exclusion test; quantitative immunocytochemistry; BrdU; XTT; TUNEL assays; and ELISA. Administration of benzene promoted the expression of apoptosis markers and reduced cell viability, all measured markers of proliferation, the release of steroid hormones and IGF-I. Overexpression of miR-152 was associated with increased cell viability, proliferation, progesterone and IGF-I release and reduced apoptosis and estradiol output. Moreover, miR-152 mitigated or prevented the effects of benzene on all the measured parameters in addition to estradiol release. The present observations suggest the toxic effect of benzene and the stimulatory influence of miR-152 on ovarian cell functions. Moreover, this is the first demonstration of the ability of miRNAs to mitigate and prevent the reproductive toxicity of benzene.
Subject(s)
Apoptosis , Benzene , Cell Proliferation , Cell Survival , Granulosa Cells , MicroRNAs , Animals , MicroRNAs/genetics , MicroRNAs/metabolism , Female , Granulosa Cells/drug effects , Granulosa Cells/metabolism , Apoptosis/drug effects , Swine , Cell Survival/drug effects , Benzene/toxicity , Cell Proliferation/drug effects , Cells, Cultured , Progesterone , Estradiol , Insulin-Like Growth Factor I/genetics , Insulin-Like Growth Factor I/metabolism , Environmental Pollutants/toxicityABSTRACT
The objective of this study was to elucidate the intricate interplay among miR-105-1, kisspeptin, and their synergistic influence on basic ovarian granulosa cell functions. The effects of miR-105-1 mimics or miR-105-1 inhibitor, kisspeptin (0, 1, and 10 ng/ml), and its combinations with miR-105-1 mimics on porcine granulosa cells were assessed. The expression levels of miR-105-1, viability, proliferation (accumulation of PCNA, cyclin B1, XTT-, and BrdU-positive cells), apoptosis (accumulation of bcl-2, bax, caspase 3, p53, TUNEL-positive cells), proportion of kisspeptin-positive cells, and the release of steroid hormones and IGF-I were analyzed. Transfection of cells with miR-105-1 mimics promoted cell viability and proliferation, the occurrence of kisspeptin, and the release of progesterone and IGF-I; in contrast, miR-105-1 mimics inhibited apoptosis and estradiol output. MiR-105-1 inhibitor had the opposite effect. Kisspeptin amplified the expression of miR-105-1, cell viability, proliferation, steroid hormones, and IGF-I release and reduced apoptosis. Furthermore, the collaborative action of miR-105-1 mimics and kisspeptin revealed a synergistic relationship wherein miR-105-1 mimics predominantly supported the actions of kisspeptin, while kisspeptin exhibited a dual role in modulating the effects of miR-105-1 mimics. These findings not only affirm the pivotal role of kisspeptin in regulating basic ovarian cell functions but also represent the inaugural evidence underscoring the significance of miR-105-1 in this regulatory framework. Additionally, our results show the ability of kisspeptin to promote miR-105-1 expression and the ability of miR-105-1 to promote the occurrence and effects of kisspeptin and, therefore, indicate the existence of the self-stimulating kisspeptin-miR-105-1 axis.
Subject(s)
Apoptosis , Cell Proliferation , Cell Survival , Granulosa Cells , Kisspeptins , MicroRNAs , Animals , MicroRNAs/metabolism , MicroRNAs/genetics , Female , Granulosa Cells/metabolism , Granulosa Cells/drug effects , Kisspeptins/metabolism , Kisspeptins/genetics , Swine , Cell Survival/physiology , Cell Survival/drug effects , Insulin-Like Growth Factor I/metabolism , Cells, Cultured , Progesterone/metabolism , Progesterone/pharmacology , Estradiol/pharmacology , Estradiol/metabolismABSTRACT
The present experiments are aimed to examine the effect of copper nanoparticles supported on charcoal (CuNPs/C), growth factor betacellulin (BTC) and their interrelationships in the control of ovarian cell functions. Porcine ovarian granulosa cells were cultured in the presence of CuNPs/C (0, 1, 10 or 100 ng/ml), BTC (100 ng/ml) and the combination of both, CuNPs/C + BTC. Markers of cell proliferation (BrDU incorporation), of the S-phase (PCNA) and G-phase (cyclin B1) of the cell cycle, markers of extrinsic (nuclear DNA fragmentation) and cytoplasmic/mitochondrial apoptosis (bax and caspase 3), and the release of progesterone and estradiol were assessed by BrDU test, TUNEL, quantitative immunocytochemistry and ELISA. Both CuNPs/C and BTC, when added alone, increased the expression of all the markers of cell proliferation, reduced the expression of all apoptosis markers and stimulated progesterone and estradiol release. Moreover, BTC was able to promote the CuNPs/C action on the accumulation of PCNA, cyclin B1, bax and estradiol output. These observations demonstrate the stimulatory action of both CuNPs/C and BTC on ovarian cell functions, as well as the ability of BTC to promote the action of CuNPs/C on ovarian cell functions.
Subject(s)
Nanoparticles , Progesterone , Female , Swine , Animals , Cyclin B1/metabolism , Progesterone/pharmacology , Charcoal/metabolism , Charcoal/pharmacology , Proliferating Cell Nuclear Antigen/metabolism , bcl-2-Associated X Protein/metabolism , Betacellulin/metabolism , Betacellulin/pharmacology , Bromodeoxyuridine/metabolism , Bromodeoxyuridine/pharmacology , Granulosa Cells , Estradiol/pharmacology , Cell Proliferation , Apoptosis , Cells, Cultured , Insulin-Like Growth Factor I/metabolismABSTRACT
The existing knowledge of the involvement of vinculin (VCL) in the control of ovarian cell functions is insufficient. To understand the role of VCL in the control of basic porcine ovarian granulosa cell functions, we decreased VCL activity by small interfering RNA (VCL siRNA). The expression of VCL, accumulation of VCL protein, cell viability, proliferation (accumulation of PCNA and cyclin B1), proportion of proliferative active cells, apoptosis (accumulation of bax, caspase 3, p53, antiapoptotic marker bcl2, and bax/bcl-2 ratio), DNA fragmentation, and release of steroid hormones and IGF-I were analyzed by RTâqPCR, Trypan blue exclusion test, quantitative immunocytochemistry, XTT assay, TUNEL assay, and ELISA. The suppression of VCL activity inhibited cell viability, the accumulation of the proliferation-related proteins PCNA and cyclin B1, the antiapoptotic protein bcl2, and the proportion of proliferative active cells. Moreover, VCL siRNA inhibited the release of progesterone, estradiol, and IGF-1. VCL siRNA increased the proportion of the proapoptotic proteins bax, caspase 3, p53, the proportion of DNA fragmented cells, and stimulated testosterone release. Taken together, the present study is the first evidence that inhibition of VCL suppresses porcine granulosa cell functions. Moreover, the results suggest that VCL can be a potent physiological stimulator of ovarian functions.
Subject(s)
Progesterone , Tumor Suppressor Protein p53 , Female , Swine , Animals , Cyclin B1/metabolism , Cyclin B1/pharmacology , Caspase 3/genetics , Caspase 3/metabolism , Proliferating Cell Nuclear Antigen/genetics , Proliferating Cell Nuclear Antigen/metabolism , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolism , Vinculin/genetics , Vinculin/metabolism , Progesterone/pharmacology , Apoptosis , Proto-Oncogene Proteins c-bcl-2/metabolism , Cell Proliferation , RNA, Small Interfering/genetics , RNA, Small Interfering/pharmacology , Cells, Cultured , Insulin-Like Growth Factor I/metabolismABSTRACT
Mycotoxins are produced by more than one hundred fungi and produce secondary metabolites that contaminate various agricultural commodities, especially rice and corn. Their presence in the food chain is considered a serious problem worldwide. In recent years, a link between exposure to mycotoxins and impaired fertility has been suggested. Consequently, it has become vital to investigate the interactive effects of these mycotoxins on ovarian function. In this study, we investigated the intergenerational effects of the mycotoxin fumonisin B1 (FB1) on ovarian structure and function. Virgin Wistar albino female rats were separated into control and FB1 treatment groups and examined from day 6 of pregnancy until delivery (20 and 50 mg/kg b.w./day). The obtained female rats of the first (F1) and second generations (F2) were euthanized at 4 weeks of age, and ovary samples were collected. We found that the ovary weight index increased with the high dose of the treatment (50 mg/kg b.w./day) among both F1 and F2, in a manner similar to that observed in polycystic ovary syndrome. As expected, FB1 at a high dose (50 mg/kg b.w.) reduced the number of primordial follicles in F1 and F2, leading to an accelerated age-related decline in reproductive capacity. Moreover, it reduced the fertility rate among the F1 female rats by affecting follicle growth and development, as the number of secondary and tertiary follicles decreased. Histopathological changes were evidenced by the altered structures of most of the growing follicle oocytes, as revealed by a thinning irregular zona pellucida and pyknosis in granulosa cells. These findings are concomitant with steroidogenesis- and folliculogenesis-related gene expression, as evidenced by the decrease in CYP19 activity and estrogen receptor beta (ESR2) gene expression. Additionally, GDF-9 mRNA levels were significantly decreased, and IGF-1 mRNA levels were significantly increased. However, the results from the ovaries of the F2 treatment groups were different and unexpected. While there was no significant variation in CYP19 activity compared to the control, the ESR2 significantly increased, leading to stereological and histopathological changes similar to those of the control, except for some altered follicles. The hallmark histological feature was the appearance of vacuolar structures within the oocyte and between granulosa cell layers. Interestingly, the autophagic marker LC3 was significantly increased in the F2 offspring, whereas this protein was significantly decreased in the F1 offspring. Therefore, we suggest that the promotion of autophagy in the ovaries of the F2 offspring may be considered a recovery mechanism from the effect of prenatal FB1 exposure. Thus, autophagy corrected the effect of FB1 during the early life of the F1 female rats, leading to F2 offspring with ovarian structure and function similar to those of the control. However, the offspring, treated female rats may experience early ovarian aging because their ovarian pool was affected.
ABSTRACT
The aim of the current work was to examine for the first time the nephropreventive capacity of Ephedra alata seed extract (E) against maternal exposure to acephate in rat offspring. The in vivo results revealed that E. alata supplementation for 28 days (40 mg/kg b.w.) significantly attenuated the nephrotoxicity in adult offspring induced by acephate. In fact, it decreased the levels of creatinine and uric acid and increased the albumin content compared to the intoxicated group. The in utero studies showed that E. alata inhibited the renal oxidative stress generated by acephate exposure by reducing lipid peroxidation and enhancing antioxidant biomarker activities (GSH, CAT, and SOD). The inhibition of DNA fragmentation and the improvement of the ultrastructural changes highlighted the prophylactic effect of E. alata in renal tissue. Additionally, the immunofluorescence study showed the upregulation of LC3 gene expression, suggesting the capacity of E. alata extract to stimulate autophagic processes as a protective mechanism. Molecular docking analysis indicated that hexadecasphinganine, the major compound in E. alata, has a higher affinity toward the Na+/K+-ATPase, epithelial sodium channel (ENaC), and sodium hydrogen exchanger 3 (NHE3) genes than acephate. Hexadecasphinganine could be considered a potential inhibitor of the activity of these genes and therefore exerted its preventive capacity. The obtained findings confirmed that E. alata seed extract exerted nephropreventive capacities, which could be related to its bioactive compounds, which possess antioxidant activities.
ABSTRACT
Fumonisin B1 (FB1), a mycotoxin produced by Fusarium verticillioides, is one of the most common pollutants in natural foods and agricultural crops. It can cause chronic and severe health issues in humans and animals. The aim of this study was to evaluate the transgenerational effects of FB1 exposure on the structure and function of the kidneys in offspring. Virgin female Wistar rats were randomly divided into three groups: group one (control) received sterile water, and groups two and three were intragastrically administered low (20 mg/kg) and high (50 mg/kg) doses of FB1, respectively, from day 6 of pregnancy until delivery. Our results showed that exposure to either dose of FB1 caused histopathological changes, such as atrophy, hypercellularity, hemorrhage, calcification, and a decrease in the glomerular diameter, in both the first and second generations. The levels of the antioxidant markers glutathione, glutathione S-transferase, and catalase significantly decreased, while malondialdehyde levels increased. Moreover, autophagy was induced, as immunofluorescence analysis revealed that LC-3 protein expression was significantly increased in both generations after exposure to either dose of FB1. However, a significant decrease in methyltransferase (DNMT3) protein expression was observed in the first generation in both treatment groups (20 mg/kg and 50 mg/kg), indicating a decrease in DNA methylation as a result of early-life exposure to FB1. Interestingly, global hypomethylation was also observed in the second generation in both treatment groups despite the fact that the mothers of these rats were not exposed to FB1. Thus, early-life exposure to FB1 induced nephrotoxicity in offspring of the first and second generations. The mechanisms of action underlying this transgenerational effect may include oxidative stress, autophagy, and DNA hypomethylation.
Subject(s)
Fumonisins , Mycotoxins , Humans , Rats , Female , Animals , Mycotoxins/toxicity , DNA Methylation , Rats, Wistar , Fumonisins/toxicity , Oxidative Stress , Autophagy , DNAABSTRACT
The action of buckwheat, rooibos and vitex on healthy female reproductive systems, as well as their ability to mitigate the reproductive toxicity of environmental contaminant toluene have not yet been examined. We analysed the influence of toluene (0, 10, 100 or 1000 ng/mL) with and without these plant extracts (10 µg/mL) on cultured porcine ovarian granulosa cells. Cell viability, proliferation (PCNA accumulation), apoptosis (accumulation of bax) and release of progesterone (P) and oestradiol (E) were measured. Toluene reduced ovarian cell viability and proliferation, increased apoptosis and suppressed E but not P release. Plant extracts, given alone, were also able to directly suppress some ovarian cell functions. The addition of buckwheat promoted toluene action on cell viability, proliferation and P release, but it did not modify other toluene effects. Rooibos mitigated toluene action on cell viability, proliferation and apoptosis but promoted its action on P and E. The addition of vitex mitigated all the tested toluene effects. These observations: (1) demonstrate the direct toxic influence of toluene on ovarian cells, (2) demonstrate the ability of food/medicinal plants to either promote or mitigate toluene effects and (3) suggest that vitex could be a natural protector against the suppressive effect of toluene on female reproduction.
Subject(s)
Plants, Medicinal , Toluene , Female , Swine , Animals , Toluene/toxicity , Cell Proliferation , Cells, Cultured , Granulosa Cells , Progesterone/pharmacology , Plant Extracts/pharmacology , ApoptosisABSTRACT
Angiotensin II (Ang II) is a major component of the renin-angiotensin or renin-angiotensin-aldosterone system, which is the main element found to be involved in cardiopathology. Recently, long-term metabolomics studies have linked high levels of angiotensin plasma to inflammatory conditions such as coronary heart disease, obesity, and type 2 diabetes. Monocyte/macrophage cellular function and phenotype orchestrate the inflammatory response in various pathological conditions, most notably cardiometabolic disease. An activation of the Ang II system is usually associated with inflammation and cardiovascular disease; however, the direct effect on monocyte/macrophages has still not been well elucidated. Herein, we have evaluated the cellular effects of Ang II on THP-1-derived macrophages. Ang II stimulated the expression of markers involved in monocyte/macrophage cell differentiation (e.g., CD116), as well as adhesion, cell-cell interaction, chemotaxis, and phagocytosis (CD15, CD44, CD33, and CD49F). Yet, Ang II increased the expression of proinflammatory markers (HLA-DR, TNF-α, CD64, CD11c, and CD38) and decreased CD206 (mannose receptor), an M2 marker. Moreover, Ang II induced cytosolic calcium overload, increased reactive oxygen species, and arrested cells in the G1 phase. Most of these effects were induced via the angiotensin II type 1 receptor (AT1R). Collectively, our results provide new evidence in support of the effect of Ang II in inflammation associated with cardiometabolic diseases.
ABSTRACT
The objective of this study was to examine the direct effects of the medicinal plant fennel on basic functions of ovarian cells, including proliferation, apoptosis, and release of progesterone and insulin-like growth factor I (IGFI), as well as to prevent the influence of the environmental contaminant benzene on these cells. Porcine ovarian granulosa cells were cultured with or without fennel extract alone or in combination with benzene. The expression of the proliferation marker PCNA and the apoptosis marker bax was analyzed by quantitative immunocytochemistry and enzyme-linked immunosorbent assay (ELISA). Fennel was able to promote proliferation and IGF-I release, but to suppress apoptosis and progesterone release. Benzene promoted the accumulation of both the proliferation and apoptosis markers, as well as IGF-I release, but it inhibited progesterone secretion. The presence of fennel did not prevent the effects of benzene on any of the measured parameters, while benzene prevented the effects of fennel on cell proliferation, apoptosis, and IGF-I but not progesterone output. These observations demonstrate the direct influence of fennel and benzene on basic ovarian cell functions. Furthermore, they show the inability of fennel to prevent the effects of benzene on these cells. On the other hand, the environmental contaminant benzene can block the response of ovarian cells to the medicinal plant fennel.
Subject(s)
Foeniculum , Progesterone , Female , Swine , Animals , Progesterone/pharmacology , Insulin-Like Growth Factor I/metabolism , Foeniculum/metabolism , Benzene/toxicity , Benzene/metabolism , Ovary , Granulosa Cells , Cell Proliferation , Apoptosis , Cells, CulturedABSTRACT
The aim of the present in-vitro experiments was to examine the direct influence of ghrelin and obestatin on viability, proliferation and progesterone release by human ovarian granulosa cells and their response to FSH administration. Human granulosa cells were cultured in presence of ghrelin or obestatin (both at 0, 1, 10 or 100 ng/ml) alone or in the presence of FSH (10 ng/ml). Cell viability, accumulation of proliferation markers PCNA and cyclin B1 and release of progesterone were analyzed by Trypan blue extrusion test, quantitative immunocytochemistry and ELISA. Ghrelin, obestatin and FSH up-regulated all the measured ovarian cell parameters. Moreover, both ghrelin and obestatin promoted all the stimulatory effects of FSH. The obtained results demonstrate the direct stimulatory action of ghrelin, obestatin and FSH on basic ovarian cell functions, as well as the ability of metabolic hormones to improve FSH action on human ovarian cells.
Subject(s)
Ghrelin , Progesterone , Female , Humans , Progesterone/pharmacology , Progesterone/metabolism , Ghrelin/metabolism , Granulosa Cells , Ovary , Follicle Stimulating Hormone/pharmacology , Follicle Stimulating Hormone/metabolism , Cells, Cultured , Cell Proliferation , ApoptosisABSTRACT
The aim of this in vitro study was to examine the potential effect of functional food plant extracts, namely, extracts of flaxseed (Linum usitatissimum L.), chia (Salvia hispanica) and puncture vine (Tribulus terrestris L.), on basic mare ovarian cell functions and their response to the environmental contaminant toluene. Mare granulosa cells were incubated with and without toluene (0, 0.02, 0.2 or 2.0 µg/mL) in the presence or absence of flaxseed, chia and puncture vine extracts (10 µg/mL). Markers of cell proliferation (accumulation of proliferating cell nuclear antigen, PCNA) and apoptosis (accumulation of bax), viability (Trypan blue extrusion) and the release of progesterone (P), oxytocin (OT) and prostaglandin F 2 alpha (PGF) were measured. Toluene reduced all other measured parameters except OT release. All the tested plants were able to reduce cell viability and the release of P and PGF, but they did not influence other indexes. Moreover, flaxseed mitigated toluene action on ovarian cell proliferation, apoptosis, OT and PGF, whilst puncture vine prevented and inverted toluene action on P and PGF ourput. Chia extract did not modify toluene action on any parameter. On the other hand, toluene was able to promote the inhibitory action of flaxseed on cell viability and P release and to prevent the inhibitory action of all the plant extracts on PGF release. The present study (1) is the first demonstration, that flaxseed, chia and puncture vine can directly suppress mare ovarian cell functions, (2) shows that toluene can suppress basic ovarian cell functions and modify the reproductive effect of food plants and (3) demonstrates the ability of flaxseed and puncture vine, but not of chia, to prevent some toxic effect of toluene on mare ovarian cell functions.