TISSUE & CELL
·
2025
·
IF 2.5
Guixiang Zeng,Jingjing Lian,Jiajia Shen,Yuan Shi
Objective The Klotho protein is associated with various aging-related diseases. Previous studies have shown that Klotho expression is reduced in placental and cord blood samples from growth-restricted fetuses, but the underlying mechanisms affecting fetal development remain unclear. This study investigates the effects of Klotho protein on the human placental choriocarcinoma cell line (JEG-3) and explores the mechanisms involved. Methods The JEG-3 cell line, which retains characteristics of trophoblast cells, is widely used to study pregnancy-related pathologies. Placental trophoblast cells act as “multifunctional hubs” at the maternal-fetal interface, performing transportation, endocrine regulation, and immune modulation. The GH/IGF-1 pathway plays a crucial role in embryonic growth and immune tolerance, and its abnormal activation is linked to preeclampsia and fetal growth restriction. To investigate the impacts of Klotho protein on the functional characteristics of JEG-3 cells, we designed two experimental setups concentrating on the low expression and high expression of Klotho protein. The research examined the expression levels of growth hormone (GH) and insulin-like growth factor 1 (IGF-1) in JEG-3 cells, along with changes in multiple cellular functions, under these two experimental conditions. Results qPCR and flow cytometry analyses showed that in the Klotho low - expression (KL -) tests, the levels of IGF - 1 mRNA in the KL - group were significantly elevated (P < 0.01, P < 0.05), while the levels of GH mRNA were significantly reduced (P < 0.001, P < 0.0001). The cell scratch test demonstrated that the cell migration capacity in the KL - group was improved (P < 0.05). The Cell Counting Kit - 8 (CCK8) test verified an increase in cell viability (P < 0.0001). Reactive oxygen species (ROS) detection showed a significant rise in ROS levels (P < < 0.001). The glucose uptake experiment indicated a decrease in glucose uptake in the KL - group (P < 0.0001). The human recombinant Klotho protein was over - expressed in the JEG - 3 cell line. Quantitative polymerase chain reaction (qPCR) and flow cytometry analyses disclosed that in the rKL + group, the levels of IGF - 1 were lowered and the levels of GH were raised (P < 0.05, P < 0.05). The cell scratch test showed that the cell migration rate in the rKL + group was diminished (P < 0.01). The CCK8 test confirmed a decrease in cell viability (P < 0.0001). The ROS detection indicated a significant decrease in ROS levels (P < 0.05). The glucose uptake experiment showed a increase in glucose uptake (P < 0.0001). Conclusions Klotho protein affects the expression of GH and IGF-1 in JEG-3 cells, leading to the dysfunction of GH/IGF-1 axis, which subsequently impacts the proliferation, migration, anti-inflammatory response, and glucose metabolism of placental trophoblast cells, ultimately affecting placental function and fetal intrauterine development. Summary sentence Reduced Klotho protein expression in JEG-3 cells leads to increased IGF-1, decreased GH, enhanced cell proliferation and migration, elevated inflammatory responses, and metabolic imbalances, potentially contributing to placental dysfunction in growth-restricted fetuses.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Functional Foods
·
2023
·
IF 5.6
Meng-ling Zhang,Qian Yang,Xiao-peng Li,Jia-hao Chen,Zi-jun Bi,Yang Shan,Li-fang Gao,Huan-ling Yu
Nobiletin (NOB) is one of the major polymethoxyflavone in citrus peels, which has antioxidant activities. The purpose of this study was to investigate the effects of NOB on hypoxia-induced placental oxidative stress and its mechanism. In vitro experiments, JEG-3 hypoxic model was constructed, and treated with 100 μM NOB. In vivo experiment, pregnant rats were injected intraperitoneally with 50 mg/kg body weight NOB daily, and the placental hypoxia model was established by RUPP. NOB hampered the reducing of reducing power in hypoxic cells, promoted NADPH to provide hydrogen donors to regenerate antioxidants GPX and GSH, thereby promoting the decomposition of ROS. NOB alleviated the adverse reaction of hypoxic placental one-carbon metabolism disorder by regulating the levels of homocysteine, DNMTs and TETs. Moreover, NOB suppressed offspring brain damage induced by RUPP. Taken together, NOB attenuated hypoxia-induced placental redox perturbation and subsequent offspring brain injury by modulating one-carbon metabolism.
研究领域:
药理学
·
妇产科学
·
神经科学
✓ 已验证
产品关联
DOI
高影响因子
EXPERIMENTAL CELL RESEARCH
·
2025
·
IF 3.5
Xiaodan Chu,Xuan Chen,Man Guo,Xinyue Li,Zhihai Qu,Peiling Li
Insufficient trophoblast cell infiltration is implicated in the progression of preeclampsia (PE). The immunoglobulin superfamily member 8 (IGSF8) has been shown to promote cell migration, invasion, and epithelial mesenchymal transition (EMT). However, the specific impact of IGSF8 on trophoblast cells in PE has not been definitively demonstrated. To address this, placental tissues from PE patients and normal subjects was collected. A PE-like rat model was established by administering L-NAME (60 mg/kg) intragastrically to pregnant rats from the 10th to the 19th day of gestation. Knockdown and overexpression plasmids of IGSF8 were transfected into JEG-3 cells for further experiments. Clinical samples indicated impaired spiral artery remodeling, and high IGSF8 expression in the placental tissues of PE patients. PE rats exhibited increased mean arterial pressure, elevated 24-h urine protein levels, higher abortion rates, and decreased placental and fetal weight compared to rats of sham group. Failure of physiological transformation of spiral arteries was observed in PE rats, along with increased IGSF8 expression. IGSF8 overexpression inhibited JEG-3 cell migration, invasion and EMT, as well as reduced release of VEGF in JEG-3 cells, impairing HUVEC tube formation. mRNA-sequencing analysis of JEG-3 cells transfected with shIGSF8 showed differentially expressed genes related to angiogenesis, and mesenchymal cell differentiation, with IGSF8 knockdown being associated with the activation of pathways involved in blood vessel development and cell migration. Overall, this study suggests that IGSF8 plays a role in the development of PE and provides new insights for potential treatments.
研究领域:
妇产科学
·
分子生物学
·
细胞生物学
·
病理生理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food & Nutrition Research
·
2021
·
IF 3.894
Mengling Zhang,Jian Liu,Rui Zhang,Zengenni Liang,Shenghua Ding,Huanling Yu,Yang Shan
Background Hypoxia is associated with abnormal cell apoptosis in trophoblast cells, which causes fetal growth restriction and related placental pathologies. Few effective methods for the prevention and treatment of placenta-related diseases exist. Natural products and functional foods have always been a rich source of potential anti-apoptotic drugs. Nobiletin (NOB), a hexamethoxyflavonoid derived from the citrus pomace, shows an anti-apoptotic activity, which is a non-toxic constituent of dietary phytochemicals approved by the Food and Drug Administration. However, their effects on hypoxia-induced human trophoblast cells have not been fully studied. Objective The aim of this study was to investigate the protective effects of NOB on hypoxia-induced apoptosis of human trophoblast JEG-3 and BeWo cells, and their underlying mechanisms. Design First, the protective effect of NOB on hypoxia-induced apoptosis of JEG-3 and BeWo cells was studied. Cell viability and membrane integrity were determined by CCK-8 assay and lactate dehydrogenase activity, respectively. Real Time Quantitative PCR (RT-qPCR) and Western blot analysis were used to detect the mRNA and protein levels of HIF1α. Propidium iodide (PI)-labeled flow cytometry was used to detect cell cycle distribution. Cell apoptosis was detected by flow cytometry with Annexin V-FITC and PI double staining, and the expression of apoptosis marker protein cl-PARP was detected by Western blot analysis. Then, the molecular mechanism of NOB against apoptosis was investigated. Computer molecular docking and dynamics were used to simulate the interaction between NOB and p53 protein, and this interaction was verified in vitro by Ultraviolet and visible spectrum (UV-visible spectroscopy), fluorescence spectroscopy and circular dichroism. Furthermore, the changes in the expression of p53 signaling pathway genes and proteins were detected by RT-qPCR and Western blot analysis, respectively. Results Hypoxia treatment resulted in a decreased cell viability and cell membrane integrity in JEG-3 and BeWo cell lines, and an increased expression of HIF1α, cell cycle arrest in the G1 phase, and massive cell apoptosis, which were alleviated after NOB treatment. Molecular docking and dynamics simulations found that NOB spontaneously bonded to human p53 protein, leading to the change of protein conformation. The intermolecular interaction between NOB and human p53 protein was further confirmed by UV-visible spectroscopy, fluorescence spectroscopy and circular dichroism. After the treatment of 100 μM NOB, a down-regulation of mRNA and protein levels of p53 and p21 and an up-regulation of BCL2/BAX mRNA and protein ratio were observed in JEG-3 cells; however, there was also a down-regulation of mRNA and protein levels observed for p53 and p21 in BeWo cells after the treatment of NOB. The BCL2/BAX ratio of BeWo cells did not change after the treatment of 100 μM NOB. Conclusion NOB attenuated hypoxia-induced apoptosis in JEG-3 and BeWo cell lines and might be a potential functional ingredient to prevent pregnancy-related diseases caused by hypoxia-induced apoptosis. These findings would also suggest the exploration and utilization of citrus resources, and the development of citrus industry.
研究领域:
药理学
·
细胞生物学
·
分子生物学
·
妇产科学
✓ 已验证
DOI
PubMed
高影响因子
REPRODUCTION FERTILITY AND DEVELOPMENT
·
2021
·Vol. 33
·期 12
·700-712
·
IF 2.311
Xin Zhan,Yiran Xie,Liping Sun,Qi Si,Hongkai Shang,Xin Zhan,Yiran Xie,Liping Sun,Qi Si,Hongkai Shang
This study explored the molecular mechanism underlying the effects of dexamethasone (DEX, 1 µM) on glucose transporters (GLUT) in JEG-3 human placental choriocarcinoma cells. JEG-3 cells were treated with DEX, an expression plasmid encoding human glucocorticoid receptor α (GRα), pcDNA3.1-GRα, GRα short interference (si) RNA, LY294002, xanthine oxidase (XO)/hypoxanthine (HX), rapamycin, insulin-like growth factor (IGF) 1, N-acetylcysteine (NAC) or phosphatidic acid (PA), and cell proliferation, apoptosis, mitochondrial membrane potential (MMP), human chorionic gonadotrophin (hCG) content, human placental lactogen (hPL) content, glucose uptake, reactive oxygen species levels and signalling pathway modulation were evaluated. Treatment of JEG-3 cells with DEX (1 µM), GRα siRNA, LY294002 (50 µM), XO/HX (7.2 µM/36 nM) or rapamycin (80 nM) inhibited cell proliferation, induced apoptosis, significantly decreased MMP and hCG and hPL content and increased ROS levels. In addition, glucose uptake was decreased through downregulation of the mRNA and protein expression of GRα, GLUT1 and GLUT3. Treatment of JEG-3 cells with GRα siRNA, LY294002, XO/HX or rapamycin inhibited phosphorylation of phosphatidylinositol 3-kinase (PI3K), Akt, glycogen synthase kinase 3 and mammalian target of rapamycin (mTOR) and induced the phosphorylation of AMP-activated protein kinase (AMPK) and tuberous sclerosis complex 2. The effects of GRα overexpression and IGF1 (100 nM), NAC (5 nM) or PA (100 µM) treatment on JEG-3 cells contrasted with those of DEX treatment. DEX blocked glucose uptake by downregulating GRα expression, which reduced GLUT1 and GLUT3 mRNA and protein expression, which, in turn, may have inhibited the PI3K/AKT/mTOR pathway and activated the ROS/AMPK pathway.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）