Roles of vitamin D and its receptor in the proliferation and apoptosis of luteinised granulosa cells in the goat

2020 ◽  
Vol 32 (3) ◽  
pp. 335 ◽  
Author(s):  
Xiaolei Yao ◽  
Zhibo Wang ◽  
M. A. El-Samahy ◽  
Caifang Ren ◽  
Zifei Liu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Vitamin D ◽  
Granulosa Cells ◽  
Expression Patterns ◽  
Follicular Development ◽  
Proliferation Rate ◽  
Control Group ◽  
Dependent Manner ◽  
Proliferation And Apoptosis ◽  
Dose Dependent

The objective of this study was to investigate the dose-dependent effect of 1α,25-(OH)2VD3 (Vit D3) on invitro proliferation of goat luteinised granulosa cells (LGCs) and to determine the underlying mechanisms of its action by overexpressing and silencing vitamin D receptor (VDR) in LGCs. Results showed that VDR was prominently localised in GCs and theca cells (TCs) and its expression increased with follicle diameter, but was lower in atretic follicles than in healthy follicles. The proliferation rate of LGCs was significantly higher in the Vit D3-treated groups than in the control group, with the highest proliferation rate observed in the 10nM group; this was accompanied by changes in the expression of cell cycle-related genes. These data indicate that Vit D3 affects LGC proliferation in a dose-dependent manner. Contrary to the VDR knockdown effects, its overexpression upregulated and downregulated cell cycle- and apoptosis-related genes respectively; moreover, supplementation with 10nM of Vit D3 significantly enhanced these effects. These results suggest that changes in VDR expression patterns in LGCs may be associated with follicular development by regulation of cell proliferation and apoptosis. These findings will enhance the understanding of the roles of Vit D3 and VDR in goat ovarian follicular development.

scholarly journals Neuropeptide Y regulates proliferation and apoptosis in granulosa cells in a follicular stage-dependent manner

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Yoko Urata ◽  
Reza Salehi ◽  
Patricia D. A. Lima ◽  
Yutaka Osuga ◽  
Benjamin K. Tsang
Keyword(s):  
Neuropeptide Y ◽  
Expression Pattern ◽  
Granulosa Cells ◽  
Signal Intensity ◽  
Follicular Development ◽  
Mitogen Activated Protein Kinase ◽  
Dependent Manner ◽  
Npy Receptor ◽  
Proliferation And Apoptosis ◽  
Ovarian Follicular Development

Abstract Background The complex regulatory mechanism involved in ovarian follicular development is not completely understood. Neuronal neuropeptide Y (NPY) is involved in the regulation of feeding behavior, energy homeostasis, and reproduction behavior, while its function in ovarian follicular development is not clear. The objective of this study was to investigate if and how NPY regulates follicle development in the ovary. Methods All experiments were performed using Sprague Dawley rats. To understand NPY expression pattern at different stages of follicular development, NPY content was assessed using immunohistochemistry in individual follicles. NPY and its receptors expression pattern were evaluated in granulosa cells isolated from preantral (PA), early antral (EA) and late antral follicles (LAF). The influence of NPY on granulosa cell proliferation and apoptosis were further assessed in vitro, using Ki67- and TUNEL-positivity assays. To investigate whether NPY induced-proliferation in EA granulosa cells is mediated through the activation of NPY receptor Y5 (NPY5R) and Mitogen-activated protein kinase (MEK) signal pathway, EA granulosa cells were treated with NPY5R antagonist (CGP71683) and MEK inhibitors (PD98059 and U0126), and Ki67-positive cells were assessed. Results NPY protein expression was follicular stage-dependent and cell type-specific. NPY signal intensity in EA was higher than those in PA and LAF. Antral granulosa cells showed the highest signal intensity compared to mural granulosa cells, cumulus cells and theca cells. Granulosa cells NPY protein content and mRNA abundance were higher in EA than in LAF. NPY receptor contents in granulosa cells were follicular stage-dependent. While NPY reduced apoptosis of EA granulosa cells, it increased the proliferation through NPY5R and MEK pathway. In contrast, in LAF granulosa cells, NPY reduced proliferation and increased the number of apoptotic cells, with no significant effects on PA granulosa cells. Conclusion This study is the first to evaluate the intraovarian role of NPY in granulosa cells at various stage of follicular development. These results indicate that NPY regulates granulosa cells proliferation and apoptosis in a follicular stage-dependent and autocrine manner. NPY may play a role in pathogenesis of ovarian follicular disorders.

Fibronectin production by chicken granulosa cells in vitro: effect of follicular development

1992 ◽  
Vol 127 (5) ◽  
pp. 466-470 ◽  
Author(s):  
Elikplimi K Asem ◽  
Jacqueline A Carnegie ◽  
Benjamin K Tsang
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
Ovarian Granulosa Cells ◽  
Preovulatory Follicles ◽  
Vitro Effect ◽  
Dose Dependent

In vitro studies were conducted to investigate the role of chicken ovarian granulosa cells in the production of fibronectin, a component of the basal lamina of ovarian follicles. Collagenase dispersed granulosa cells obtained from the first (F1; about 35 mm in diameter) and third (F3; 15–20 mm in diameter) largest preovulatory follicles, as well as from a pool of small yellow follicles (SF; 6–10 mm in diameter), were incubated in serum-free medium-199 for 24 to 96 h in the absence and presence of luteinizing hormone (LH) or forskolin. Fibronectin secreted in the medium was quantitated by enzyme linked immunosorbent assay. Basal fibronectin production (which increased with the duration of incubation) was significantly greater (p<0.001) in granulosa cells derived from mature follicle (F1) than in F3 or SF cells. Both LH and forskolin stimulated fibronectin production in SF and F3 cells in a dose-dependent manner; however, they were without effect in F1 cells. The magnitude of increase in fibronectin production elicited by LH or forskolin was greater in SF cells than in F3 cells. The cytoplasm of cultured granulosa cells taken at all stages of follicular development stained positively for fibronectin. These findings indicate that chicken granulosa cells produce fibronectin. This ability is acquired early in follicular development and the stimulatory effect of the gonadotropin (LH) diminished as the follicle approached ovulation.

scholarly journals Anti-Proliferation Activity of a Decapeptide from Perinereies aibuhitensis toward Human Lung Cancer H1299 Cells

Marine Drugs ◽  
2019 ◽  
Vol 17 (2) ◽  
pp. 122 ◽  
Author(s):  
Shuoqi Jiang ◽  
Yinglu Jia ◽  
Yunping Tang ◽  
Die Zheng ◽  
Xingbiao Han ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cell Cycle ◽  
Human Lung ◽  
Human Lung Cancer ◽  
Cycle Phase ◽  
Control Group ◽  
Dependent Manner ◽  
Apoptotic Protein ◽  
Perinereis Aibuhitensis ◽  
Dose Dependent

Perinereis aibuhitensis peptide (PAP) is a decapeptide (Ile-Glu-Pro-Gly-Thr-Val-Gly-Met-Met-Phe, IEPGTVGMMF) with anticancer activity that was purified from an enzymatic hydrolysate of Perinereis aibuhitensis. In the present study, the anticancer effect of PAP on H1299 cell proliferation was investigated. Our results showed that PAP promoted apoptosis and inhibited the proliferation of H1299 cells in a time- and dose-dependent manner. When the PAP concentration reached 0.92 mM, more than 95% of treated cells died after 72 h of treatment. Changes in cell morphology were further analyzed using an inverted microscope and AO/EB staining and flow cytometry was adopted for detecting apoptosis and cell cycle phase. The results showed that the early and late apoptosis rates of H1299 cells increased significantly after treatment with PAP and the total apoptosis rate was significantly higher than that of the control group. Moreover, after treatment with PAP, the number of cells in the S phase of cells was significantly reduced and the ability for the cells to proliferate was also reduced. H1299 cells were arrested in the G2/M phase and cell cycle progression was inhibited. Furthermore, the results of western blotting showed that nm23-H1 and vascular endothelial growth factor (VEGF) protein levels decreased in a dose-dependent manner, while the pro-apoptotic protein and anti-apoptotic protein ratios and the level of apoptosis-related caspase protein increased in a dose-dependent manner. In conclusion, our results indicated that PAP, as a natural marine bioactive substance, inhibited proliferation and induced apoptosis of human lung cancer H1299 cells. PAP is likely to be exploited as the functional food or adjuvant that may be used for prevention or treatment of human non-small cell lung cancer in the future.

scholarly journals Effect of large dietary doses of ß-carotene on plasma retinoid AND ß-carotene levels and ON progesterone production in the granulosa cells of Japanese quail

2000 ◽  
Vol 48 (1) ◽  
pp. 81-87 ◽  
Author(s):  
Gabriella Ágota ◽  
L. Bárdos ◽  
A. Pusztai
Keyword(s):  
Japanese Quail ◽  
Granulosa Cells ◽  
Basal Diet ◽  
Retinyl Palmitate ◽  
Control Group ◽  
Dependent Manner ◽  
Blood Samples ◽  
Progesterone Secretion ◽  
Dose Dependent ◽  
Control Group Group

An experiment was conducted to study the effect of large-dose (-carotene supplementation on blood retinoid and (-carotene levels as well as on the progesterone secretion of the granulosa cells in Japanese quail. Laying quails were assigned to three dietary groups. The control group (Group C) received the basal diet (laying feed containing 9000 IU vitamin A/kg). In the treated groups (Groups BC1 and BC2) the basal diet was supplemented with 102and 103mg/kg (-carotene (BC), respectively. At the end of the two-week feeding period, 10 birds from each group were euthanised. Blood samples were analysed for retinol, retinyl palmitate and (-carotene concentrations. Granulosa cells were isolated from ovarian follicles (F1and F2), and PMSG-inducedin vitroprogesterone (P4) secretion was measured. Similar retinol concentrations were found in both (-carotene supplemented groups, indicating saturation of the retinol-transporting system. (-carotene supplementation was accompanied by hypercarotenaemia, but did not increase the retinyl palmitate levels in the blood. PMSG-induced P4production of the granulosa cells decreased significantly in Groups BC1 and BC2 in a dose-dependent manner.

scholarly journals MiR-23a induced the activation of CDC42/PAK1 pathway and cell cycle arrest in human ovarian granulosa cells by targeting FGD4

2019 ◽  
Author(s):  
Ji Lin ◽  
Huijuan Huang ◽  
Liheng Lin ◽  
Weiwei Li ◽  
Jianfen Huang
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Granulosa Cells ◽  
Follicular Development ◽  
Inhibitory Effect ◽  
Luciferase Reporter ◽  
Ovarian Granulosa Cells ◽  
Cycle Arrest ◽  
Proliferation And Apoptosis ◽  
Healthy Control

Abstract Background: MiRNAs play important roles in the development of ovarian cancer, activation of primitive follicles, follicular development, oocyte maturation and ovulation. In the present study, we investigated the specific role of miR-23a in human ovarian granulosa cells. Results: Downregulation of miR-23a was observed in in serum of PCOS patients compared with the healthy control, suggesting the inhibitory effect of miR-23a in PCOS. MiR-23a was positively correlated with Body Mass Index (BMI) and negatively correlated with Luteinizing hormone (LH) of PCOS patients. MiR-23a mimic inhibited the proliferation and promoted apoptosis of human ovarian granulosa cells. In addition, flow cytometry assay confirmed that miR-23a blocked cell cycle on G0/G1 phase. MiR-23a inhibitor showed opposite results. Furthermore, double luciferase reporter assay proved that miR-23a could bind to the 3’UTR of FGD4 directly through sites predicted on Target Scan. FGD4 level was significantly suppressed by miR-23a mimic, but was significantly enhanced by miR-23a inhibitor. We further proved that miR-23a increased the expression of activated CDC42 (GTP bround) and p-PAK-1, suggesting that miR-23a induced cell cycle arrest through CDC42/PAK1 pathway. Conclusions: In conclusion, our study reveals that miR-23a participates in the regulation of proliferation and apoptosis of ovarian granulosa cells through target FGD4, which may have potential for clinical diagnosis and treatment of PCOS patients.

scholarly journals Ginsenoside-Rb1 Protects Hypoxic- and Ischemic-Damaged Cardiomyocytes by Regulating Expression of miRNAs

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Xu Yan ◽  
Jinrong Xue ◽  
Hongjin Wu ◽  
Shengqi Wang ◽  
Yuna Liu ◽  
...  
Keyword(s):  
Neonatal Rat ◽  
Control Group ◽  
Dependent Manner ◽  
The Novel ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes ◽  
Proliferation And Apoptosis ◽  
Novel Strategy ◽  
Dose Dependent

Ginsenoside (GS-Rb1) is one of the most important active compounds of ginseng, with extensive evidence of its cardioprotective properties. However, the miRNA mediated mechanism of GS-Rb1 on cardiomyocytes remains unclear. Here, the roles of miRNAs in cardioprotective activity of GS-Rb1 were investigated in hypoxic- and ischemic-damaged cardiomyocytes. Neonatal rat cardiomyocytes (NRCMs) were first isolated, cultured, and then incubated with or without GS-Rb1 (2.5–40μM)in vitrounder conditions of hypoxia and ischemia. Cell growth, proliferation, and apoptosis were detected by MTT and flow cytometry. Expressions of various microRNAs were analyzed by real-time PCR. Compared with that of the control group, GS-Rb1 significantly decreased cell death in a dose-dependent manner and expressions of mir-1, mir-29a, and mir-208 obviously increased in the experimental model groups. In contrast, expressions of mir-21 and mir-320 were significantly downregulated and GS-Rb1 could reverse the differences in a certain extent. The miRNAs might be involved in the protective effect of GS-Rb1 on the hypoxia/ischemia injuries in cardiomyocytes. The effect might be based on the upregulation of mir-1, mir-29a, and mir-208 and downregulation of mir-21 and mir-320. This might provide us a new target to explore the novel strategy for ischemic cardioprotection.

scholarly journals MiR-23a induced the activation of CDC42/PAK1 pathway and cell cycle arrest in human granulosa cells by targeting FGD4

2020 ◽  
Author(s):  
Ji Lin ◽  
Huijuan Huang ◽  
Liheng Lin ◽  
Weiwei Li ◽  
Jianfen Huang
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Granulosa Cells ◽  
Follicular Development ◽  
Inhibitory Effect ◽  
Luciferase Reporter ◽  
Human Granulosa Cells ◽  
Cycle Arrest ◽  
Proliferation And Apoptosis ◽  
Healthy Control

Abstract Background MiRNAs play important roles in the development of ovarian cancer, activation of primitive follicles, follicular development, oocyte maturation and ovulation. In the present study, we investigated the specific role of miR-23a in human granulosa cells. Results Downregulation of miR-23a was observed in in serum of PCOS patients compared with the healthy control, suggesting the inhibitory effect of miR-23a in PCOS. MiR-23a was positively correlated with Body Mass Index (BMI) and negatively correlated with Luteinizing hormone (LH), Testostrone (T), Glucose (Glu) and Insulin (INS) of PCOS patients. MiR-23a mimic inhibited the proliferation and promoted apoptosis of human granulosa cells. In addition, flow cytometry assay confirmed that miR-23a blocked cell cycle on G0/G1 phase. MiR-23a inhibitor showed opposite results. Furthermore, double luciferase reporter assay proved that miR-23a could bind to the 3’UTR of FGD4 directly through sites predicted on Target Scan. FGD4 level was significantly suppressed by miR-23a mimic, but was significantly enhanced by miR-23a inhibitor. We further proved that miR-23a increased the expression of activated CDC42 (GTP bround) and p-PAK-1, suggesting that miR-23a induced cell cycle arrest through CDC42/PAK1 pathway. Conclusions In conclusion, our study reveals that miR-23a participates in the regulation of proliferation and apoptosis of ovarian granulosa cells through target FGD4, and may play a role in the pathophysiology of PCOS.

scholarly journals Apoptosis of rat hepatic stellate cells induced by diallyl trisulfide and proteomics profiling in vitro

2017 ◽  
Vol 95 (5) ◽  
pp. 463-473
Author(s):  
Yajie Zhang ◽  
Xiaoming Zhou ◽  
Lipeng Xu ◽  
Lulu Wang ◽  
Jinling Liu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Protein Expression ◽  
Hepatic Stellate Cells ◽  
Colorimetric Assay ◽  
Expression Patterns ◽  
Stellate Cells ◽  
Control Group ◽  
Dependent Manner ◽  
Diallyl Trisulfide ◽  
Proliferation And Apoptosis

Diallyl trisulfide (DATS), a major garlic derivative, inhibits cell proliferation and triggers apoptosis in a variety of cancer cell lines. However, the effects of DATS on hepatic stellate cells (HSCs) remain unknown. The aim of this study was to analyze the effects of DATS on cell proliferation and apoptosis, as well as the protein expression profile in rat HSCs. Rat HSCs were treated with or without 12 and 24 μg/mL DATS for various time intervals. Cell proliferation and apoptosis were determined using tetrazolium dye (MTT) colorimetric assay, bromodeoxyuridine (5-bromo-2′-deoxyuridine; BrdU) assay, Hoechst 33342 staining, electroscopy, and flow cytometry. Protein expression patterns in HSCs were systematically studied using 2-dimensional electrophoresis and mass spectrometry. DATS inhibited cell proliferation and induced apoptosis of HSCs in a time-dependent manner. We observed clear morphological changes in apoptotic HSCs and dramatically increased annexin V-positive – propidium iodide negative apoptosis compared with the untreated control group. Twenty-one significant differentially expressed proteins, including 9 downregulated proteins and 12 upregulated proteins, were identified after DATS administration, and most of them were involved in apoptosis. Our results suggest that DATS is an inducer of apoptosis in HSCs, and several key proteins may be involved in the molecular mechanism of apoptosis induced by DATS.

Synthesis and Biological Evaluation of Novel Heterocyclic Imines Linked Coumarin- Thiazole Hybrids as Anticancer Agents

2019 ◽  
Vol 19 (4) ◽  
pp. 557-566 ◽  
Author(s):  
Nerella S. Goud ◽  
Mahammad S. Ghouse ◽  
Jatoth Vishnu ◽  
Jakkula Pranay ◽  
Ravi Alvala ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Membrane Potential ◽  
Fluorescence Spectroscopy ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Annexin V ◽  
Biological Evaluation ◽  
Dependent Manner ◽  
Dapi Staining ◽  
Dose Dependent

Background: Human Galectin-1, a protein of lectin family showing affinity towards β-galactosides has emerged as a critical regulator of tumor progression and metastasis, by modulating diverse biological events including homotypic cell aggregation, migration, apoptosis, angiogenesis and immune escape. Therefore, galectin-1 inhibitors might represent novel therapeutic agents for cancer. Methods: A new series of heterocyclic imines linked coumarin-thiazole hybrids (6a-6r) was synthesized and evaluated for its cytotoxic potential against a panel of six human cancer cell lines namely, lung (A549), prostate (DU-145), breast (MCF-7 & MDA-MB-231), colon (HCT-15 & HT-29) using MTT assay. Characteristic apoptotic assays like DAPI staining, cell cycle, annexin V and Mitochondrial membrane potential studies were performed for the most active compound. Furthermore, Gal-1 inhibition was confirmed by ELISA and fluorescence spectroscopy. Results: Among all, compound 6g 3-(2-(2-(pyridin-2-ylmethylene) hydrazineyl) thiazol-4-yl)-2H-chromen-2- one exhibited promising growth inhibition against HCT-15 colorectal cancer cells with an IC50 value of 1.28 ± 0.14 µM. The characteristic apoptotic morphological features like chromatin condensation, membrane blebbing and apoptotic body formation were clearly observed with compound 6g on HCT-15 cells using DAPI staining studies. Further, annexin V-FITC/PI assay confirmed effective early apoptosis induction by treatment with compound 6g. Loss of mitochondrial membrane potential and enhanced ROS generation were confirmed with JC-1 and DCFDA staining method, respectively by treatment with compound 6g, suggesting a possible mechanism for inducing apoptosis. Moreover, flow cytometric analysis revealed that compound 6g blocked G0/G1 phase of the cell cycle in a dose-dependent manner. Compound 6g effectively reduced the levels of Gal-1 protein in a dose-dependent manner. The binding constant (Ka) of 6g with Gal-1 was calculated from the intercept value which was observed as 1.9 x 107 M-1 by Fluorescence spectroscopy. Molecular docking studies showed strong interactions of compound 6g with Gal-1 protein. Conclusion: Our studies demonstrate the anticancer potential and Gal-1 inhibition of heterocyclic imines linked coumarin-thiazole hybrids.

scholarly journals Effect of Synchronous Versus Sequential Regimens on the Pharmacokinetics and Biodistribution of Regorafenib with Irradiation

Pharmaceutics ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 386
Author(s):  
Tung-Hu Tsai ◽  
Yu-Jen Chen ◽  
Li-Ying Wang ◽  
Chen-Hsi Hsieh
Keyword(s):  
Stereotactic Body Radiation Therapy ◽  
In Vivo Studies ◽  
Control Group ◽  
High Dose ◽  
Dependent Manner ◽  
Hep G2 ◽  
Time Curve ◽  
Dose Dependent

This study was performed to evaluate the interaction between conventional or high-dose radiotherapy (RT) and the pharmacokinetics (PK) of regorafenib in concurrent or sequential regimens for the treatment of hepatocellular carcinoma. Concurrent and sequential in vitro and in vivo studies of irradiation and regorafenib were designed. The interactions of RT and regorafenib in vitro were examined in the human hepatoma Huh-7, HA22T and Hep G2 cell lines. The RT–PK phenomenon and biodistribution of regorafenib under RT were confirmed in a free-moving rat model. Regorafenib inhibited the viability of Huh-7 cells in a dose-dependent manner. Apoptosis in Huh-7 cells was enhanced by RT followed by regorafenib treatment. In the concurrent regimen, RT decreased the area under the concentration versus time curve (AUC)regorafenib by 74% (p = 0.001) in the RT2 Gy × 3 fraction (f’x) group and by 69% (p = 0.001) in the RT9 Gy × 3 f’x group. The AUCregorafenib was increased by 182.8% (p = 0.011) in the sequential RT2Gy × 1 f’x group and by 213.2% (p = 0.016) in the sequential RT9Gy × 1 f’x group. Both concurrent regimens, RT2Gy × 3 f’x and RT9Gy × 3 f’x, clearly decreased the biodistribution of regorafenib in the heart, liver, lung, spleen and kidneys, compared to the control (regorafenib × 3 d) group. The concurrent regimens, both RT2Gy × 3 f’x and RT9Gy × 3 f’x, significantly decreased the biodistribution of regorafenib, compared with the control group. The PK of regorafenib can be modulated both by off-target irradiation and stereotactic body radiation therapy (SBRT).

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