122 results on '"Zhao, Lijia"'
Search Results
102. Novel thermomechanical processing methods for achieving ultragrain refinement of low-carbon steel without heavy plastic deformation.
- Author
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Zhao, Lijia, Park, Nokeun, Tian, Yanzhong, Chen, Shuai, Shibata, Akinobu, and Tsuji, Nobuhiro
- Subjects
RECRYSTALLIZATION (Metallurgy) ,GRAIN refinement ,THERMODYNAMICS ,DEGRADATION of steel ,STEEL supply & demand - Abstract
Two novel two-step thermomechanical routes were developed to produce ultrafine-grained ferrite microstructures in a 10Ni–0.1C steel without high-strain deformation. Homogeneous ultrafine ferrite (UFF) structures having mean grain sizes down to 460 nm were fabricated by a total equivalent strain of 0.92 and exhibited high yield strength of 820 MPa with large uniform elongation of 10% and total elongation of 29%. The formation of UFF was attributed to two different phenomena, i.e. dynamic transformation from austenite to ferrite and dynamic recrystallization of ferrite. IMPACT STATEMENT Ultragrain refinement and remarkable strength-ductility properties can be achieved by controlling dynamic transformation and recrystallization in a thermomechanically controlled process without high-strain deformation. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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103. Downregulation of core clock gene Bmal1 attenuates expression of progesterone and prostaglandin biosynthesis-related genes in rat luteinizing granulosa cells
- Author
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Chen, Huatao, primary, Zhao, Lijia, additional, Kumazawa, Makoto, additional, Yamauchi, Nobuhiko, additional, Shigeyoshi, Yasufumi, additional, Hashimoto, Seiichi, additional, and Hattori, Masa-aki, additional
- Published
- 2013
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104. FSH induces the development of circadian clockwork in rat granulosa cells via a gap junction protein Cx43-dependent pathway
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Chen, Huatao, primary, Zhao, Lijia, additional, Chu, Guiyan, additional, Kito, Gakushi, additional, Yamauchi, Nobuhiko, additional, Shigeyoshi, Yasufumi, additional, Hashimoto, Seiichi, additional, and Hattori, Masa-aki, additional
- Published
- 2013
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105. The nuclear receptor REV- ERB α represses the transcription of growth/differentiation factor 10 and 15 genes in rat endometrium stromal cells.
- Author
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Zhao, Lijia, Isayama, Keishiro, Chen, Huatao, Yamauchi, Nobuhiko, Shigeyoshi, Yasufumi, Hashimoto, Seiichi, and Hattori, Masa‐aki
- Subjects
- *
NUCLEAR receptors (Biochemistry) , *GROWTH factors , *GENE expression , *BIOLUMINESCENCE , *STROMAL cells - Abstract
Cellular oscillators in the uterus play critical roles in the gestation processes of mammals through entraining of the clock proteins to numerous downstream genes, including growth/differentiation factor ( Gdf) 10 and Gdf15. The expression of Gdf10 and Gdf15 is significantly increased in the uterus during decidualization, but the mechanism underlying the regulation of Gdf gene expression in the uterus is poorly understood. Here, we focused on the function of the cellular oscillators in the expression of Gdf family by using uterine endometrial stromal cells ( UESCs) isolated from pregnant Per2- dLuc transgenic rats. A significant decline of Per2- dLuc bioluminescence activity was induced in in vitro decidualized UESCs, and concomitantly the expression of canonical clock genes was downregulated. Conversely, the expression of Gdf10 and Gdf15 of the Gdf was upregulated. In UESCs transfected with Bmal1-specific si RNA, in which Rev-erbα expression was downregulated, Gdf10 and Gdf15 were upregulated. However, Gdf5, Gdf7, and Gdf11 were not significantly affected by Bmal1 silencing. The expression of Gdf10 and Gdf15 was enhanced after treatment with a REV- ERB α antagonist in the presence or absence of progesterone. Chromatin immunoprecipitation- PCR analysis revealed the inhibitory effect of REV- ERB α on the expression of Gdf10 and Gdf15 in UESCs by recognizing their gene promoters. Collectively, our findings indicate that the attenuation of REV- ERB α leads to an upregulation of Gdf10 and Gdf15 in decidual cells, in which cellular oscillators are impaired. Our results provide novel evidence regarding the functions of cellular oscillators regulating the expression of downstream genes during the differentiation of UESCs. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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106. Enhanced adsorption of malachite green onto carbon nanotube/polyaniline composites
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Zeng, You, primary, Zhao, Lijia, additional, Wu, Wendong, additional, Lu, Guixia, additional, Xu, Feng, additional, Tong, Yu, additional, Liu, Wenbin, additional, and Du, Jinhong, additional
- Published
- 2012
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107. Rev-erbα regulates circadian rhythms and StAR expression in rat granulosa cells as identified by the agonist GSK4112
- Author
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Chen, Huatao, primary, Chu, Guiyan, additional, Zhao, Lijia, additional, Yamauchi, Nobuhiko, additional, Shigeyoshi, Yasufumi, additional, Hashimoto, Seiichi, additional, and Hattori, Masa-aki, additional
- Published
- 2012
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108. The Incentive Mechanism of AA Company
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Chao, Ke, primary, Zhao, Lijia, additional, and Qiao, Cai hong, additional
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- 2011
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109. Integration of the nuclear receptor REV-ERBα linked with circadian oscillators in the expressions of Alas1, Ppargc1a, and Il6 genes in rat granulosa cells.
- Author
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Chen, Huatao, Isayama, Keishiro, Kumazawa, Makoto, Zhao, Lijia, Yamauchi, Nobuhiko, Shigeyoshi, Yasufumi, Hashimoto, Seiichi, and Hattori, Masa-aki
- Subjects
NUCLEAR receptors (Biochemistry) ,CIRCADIAN rhythms ,GENE expression ,GENETIC transcription ,PROMOTERS (Genetics) - Abstract
The nuclear receptor REV-ERBα links circadian rhythms and numerous physiological processes, but its physiological role in ovaries remains largely unknown. The aim of this study was to determine the potential role of REV-ERBα in the regulation of the transcription of its putative target genes in granulosa cells (GCs) prepared from Per2-destablized luciferase ( dLuc) reporter gene transgenic rats. Alas1, Ppargc1a, and Il6 were chosen as representatives for genes analysis. A real-time monitoring system of Per2 promoter activity was performed to detect Per2-dLuc circadian oscillations. Two agonists (GSK4112, heme) and an antagonist (SR8278) of REV-ERBα as well as Rev-erbα siRNA knockdown were used to identify its target genes. Clear Per2-dLuc circadian oscillations were generated in matured GCs after synchronization with GSK4112 or SR8278. GSK4112 treatment lengthened and SR8278 treatment shortened the period of circadian oscillations in matured GCs stimulated with or without luteinizing hormone (LH). GSK4112 showed an inhibitory effect on the amplitude of circadian oscillations and caused an arrhythmic expression of canonical clock genes. SR8278 also had a subtle effect on their daily expression profiles, but the treatment resulted only in the arrhythmic expression of Rev-erbα. These findings indicate the functional biological activity of REV-ERBα in response to its ligands. Its natural ligand heme further elongated the period of circadian oscillations and alleviated their amplitudes in GCs cultured with LH. Heme treatment also repressed the expressions of clock genes, Alas1, Il6, and Ppargc1a. Rev-erbα knockdown up-regulated these transcript levels. Collectively, these data extend the recent finding to rat GCs and demonstrate that REV-ERBα represses the expressions of Alas1, Ppargc1a, and Il6, providing novel insights into the physiological significance of REV-ERBα in ovarian circadian oscillators. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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110. Enhanced adsorption of malachite green onto carbon nanotube/polyaniline composites.
- Author
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ZENg, You, Zhao, Lijia, Wu, WENdong, Lu, Guixia, Xu, FENg, Tong, Yu, Liu, WENbin, and Du, Jinhong
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MALACHITE green ,CARBON nanotubes ,ADSORPTION (Chemistry) ,POLYANILINES ,NANOCOMPOSITE materials ,ORGANIC dyes ,POLYMERIZATION - Abstract
To enhance adsorption of organic dyes like malachite green (MG) onto polymeric absorbents, we prepared carbon nanotube (CNT) filled polyaniline (PANI) composites with large surface areas by simply using entangled CNTs as porous frameworks during PANI polymerization. Adsorption behavior of the CNT/PANI composites in MG solutions was experimentally investigated and theoretically analyzed. The CNT/PANI composites exhibit much higher equilibrium adsorption capacity of 13.95 mg g
−1 at an initial MG concentration of 16 mg L−1 , increasing by 15% than the neat PANI, which is mainly attributed to large surface areas and strong CNT-PANI interactions of the composites. In addition, theoretical analyses indicate that the adsorption kinetics and the isothermal process of the composites can be well explained by using the Ho pseudosecond-order model and the Langmuir model, respectively. In light of their high MG adsorption and easy operation, the CNT/PANI composites have great potential as high-efficiency adsorbents for removal of dyes from wastewater. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013 [ABSTRACT FROM AUTHOR]- Published
- 2013
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111. Procyanidine alleviates bisphenol A-induced apoptosis in TM3 cells via the Nrf2 signaling pathway.
- Author
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Pan F, Yang W, Zhao T, Liu K, Zhao S, and Zhao L
- Abstract
Contaminated foods are a major source of bisphenol A (BPA) and are widely used in food packaging. Prolonged exposure to BPA can cause reproductive dysfunction in humans. Procyanidine (PC) is a potent natural antioxidant; however, the exact mechanism by which PC mitigates Leydig cell damage caused by BPA is unknown. In this study, the protective effect of PC against BPA-induced TM3 cell damage was investigated, and the underlying mechanism was assessed. PC treatment attenuates BPA-induced TM3 cell damage by suppressing oxidative stress and inhibiting TM3 apoptosis. In addition, PC upregulates the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream antioxidant target genes. Treatment with the NRF2 inhibitor ML385 reversed the PC-induced upregulation of the mRNA expression of these genes. Overall, PC may mitigate BPA-induced cell damage by activating the Nrf2 signaling pathway, suggesting that PC supplementation may alleviate BPA toxicity in TM3 cells., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)
- Published
- 2024
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112. Effect of dipeptidyl peptidase-4 inhibitors on tumor necrosis factor alpha levels in patients with type 2 diabetes mellitus.
- Author
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Zhao L, Meng J, Bai X, Zhang D, Yang X, Yang Y, Cai G, and Liu X
- Subjects
- Humans, Hypoglycemic Agents therapeutic use, Randomized Controlled Trials as Topic, Diabetes Mellitus, Type 2 drug therapy, Diabetes Mellitus, Type 2 blood, Dipeptidyl-Peptidase IV Inhibitors therapeutic use, Tumor Necrosis Factor-alpha blood
- Abstract
Aims: Dipeptidyl peptidase-4 inhibitors (DPP-4i) served as oral antidiabetic agents for treatment of type 2 diabetes mellitus (T2DM). Although an action on glucose homeostasis was identified, no well-rounded illustration had been established on the changes of tumor necrosis factor alpha (TNF-alpha) levels during DPP-4i treatment. This study aimed to explore the anti-inflammatory effect of DPP-4i on TNF-alpha in patients with T2DM., Methods: PubMed, Embase and Cochrane Library were systematically searched from inception to May 31, 2024. Randomized controlled trials exploring the impact of DPP-4i on TNF-alpha levels were identified. Risk of bias was assessed according to the Cochrane criteria. A fixed or random-effects model was selected to pool estimate on whether the heterogeneity was present. Subgroup analysis were performed to explore the potential factors that influenced heterogeneity. Related meta-analysis was conducted with the software of Revman 5.3 and STATA 12.0., Results: Eleven trials involving 884 participants with T2DM were included. Pooled estimates suggested that DPP-4i did not significantly modulate TNF-alpha levels (WMD, - 0.70, 95% CI - 1.94 to 0.53, P = 0.26) in T2DM. DPP-4i produced a significant effect on TNF-alpha (WMD, - 4.50 pg/mL, 95% CI - 4.68 to - 4.32, P < 0.00001) when compared to placebo, and a comparable effect was demonstrated on TNF-alpha (WMD, 0.10 pg/mL, 95% CI - 0.11 to 0.30, P = 0.35) in comparison with active agents. Estimate was stable according to the sensitivity test. Subgroup analysis revealed that heterogeneity might not correlate with baseline glycated hemoglobin (HbA1c), age or treatment duration., Conclusions: A significant effect of DPP-4i on TNF-alpha levels was present in T2DM when compared to placebo. Administration of DPP-4i produced no significant effect on TNF-alpha in comparison with active comparators. Further studies with large samples should be performed to illustrate the impact of DPP-4i on TNF-alpha levels in T2DM. Trial registration International Prospective Register for Systematic Review (PROSPERO) number: CRD42020185479., (© 2024. The Author(s).)
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- 2024
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113. Plasticity in single-crystalline Mg 3 Bi 2 thermoelectric material.
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Zhao P, Xue W, Zhang Y, Zhi S, Ma X, Qiu J, Zhang T, Ye S, Mu H, Cheng J, Wang X, Hou S, Zhao L, Xie G, Cao F, Liu X, Mao J, Fu Y, Wang Y, and Zhang Q
- Subjects
- Tensile Strength, Magnesium Compounds chemistry, Magnesium chemistry, Barium chemistry, Tellurium chemistry, Temperature, Crystallization
- Abstract
Most of the state-of-the-art thermoelectric materials are inorganic semiconductors. Owing to the directional covalent bonding, they usually show limited plasticity at room temperature
1,2 , for example, with a tensile strain of less than five per cent. Here we discover that single-crystalline Mg3 Bi2 shows a room-temperature tensile strain of up to 100 per cent when the tension is applied along the (0001) plane (that is, the ab plane). Such a value is at least one order of magnitude higher than that of traditional thermoelectric materials and outperforms many metals that crystallize in a similar structure. Experimentally, slip bands and dislocations are identified in the deformed Mg3 Bi2 , indicating the gliding of dislocations as the microscopic mechanism of plastic deformation. Analysis of chemical bonding reveals multiple planes with low slipping barrier energy, suggesting the existence of several slip systems in Mg3 Bi2 . In addition, continuous dynamic bonding during the slipping process prevents the cleavage of the atomic plane, thus sustaining a large plastic deformation. Importantly, the tellurium-doped single-crystalline Mg3 Bi2 shows a power factor of about 55 microwatts per centimetre per kelvin squared and a figure of merit of about 0.65 at room temperature along the ab plane, which outperforms the existing ductile thermoelectric materials3,4 ., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2024
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114. Two Novel Three-Dimensional Tetraphenylethylene-Based Rare Earth MOFs with Ultra-High Proton Conductivity and Performance Stability.
- Author
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Wang S, Zhao L, Sun H, Wu Y, Wang R, Zhang S, Du L, and Zhao QH
- Abstract
In this work, the two example rare earth-based metal-organic frameworks (La
III -based MOFs), Eu-ETTB and Gd-ETTB, were obtained by self-assembly. Both materials showed extremely high proton conductivity, with the proton conductivity of Eu-ETTB being 1.53×10-2 S cm-1 at 98 % relative humidity (RH) and 85 °C and that of Gd-ETTB being 2.63×10-2 S cm-1 at 98 % RH and 75 °C. This was almost the best performance observed for three-dimensional porous MOFs without post-synthetic modification and was based on milder conditions than for most materials. Furthermore, cycle test experiments and continuous work tests showed that both materials had excellent performance both in terms of stability and durability. Water vapor adsorption experiments showed that a large number of water molecules are adsorbed the hydrogen-bond network's being rebuilt by the adsorbed water molecules in the pore channel and thus optimizing the channels for proton transfer explained the MOF's high performance., (© 2022 Wiley-VCH GmbH.)- Published
- 2022
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115. Circadian clock regulates granulosa cell autophagy through NR1D1-mediated inhibition of ATG5.
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Zhang J, Zhao L, Li Y, Dong H, Zhang H, Zhang Y, Ma T, Yang L, Gao D, Wang X, Jiang H, Li C, Wang A, Jin Y, and Chen H
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- Animals, Autophagy-Related Protein 5 antagonists & inhibitors, Autophagy-Related Protein 5 genetics, Cells, Cultured, Circadian Rhythm Signaling Peptides and Proteins biosynthesis, Circadian Rhythm Signaling Peptides and Proteins genetics, Female, Granulosa Cells pathology, HEK293 Cells, Humans, Mice, Mice, Inbred C57BL, Mice, Transgenic, Nuclear Receptor Subfamily 1, Group D, Member 1 genetics, Autophagy physiology, Autophagy-Related Protein 5 biosynthesis, Circadian Clocks physiology, Circadian Rhythm physiology, Granulosa Cells metabolism, Nuclear Receptor Subfamily 1, Group D, Member 1 biosynthesis
- Abstract
Autophagy of granulosa cells (GCs) is involved in follicular atresia, which occurs repeatedly during the ovarian development cycle. Several circadian clock genes are rhythmically expressed in both rodent ovarian tissues and GCs. Nuclear receptor subfamily 1 group D member 1 (NR1D1), an important component of the circadian clock system, is involved in the autophagy process through the regulation of autophagy-related genes. However, there are no reports illustrating the role of the circadian clock system in mouse GC autophagy. In the present study, we found that core circadian clock genes ( Bmal1 , Per2 , Nr1d1 , and Dbp ) and an autophagy-related gene ( Atg5 ) exhibited rhythmic expression patterns across 24 h in mouse ovaries and primary GCs. Treatment with SR9009, an agonist of NR1D1, significantly reduced the expression of Bmal1 , Per2 , and Dbp in mouse GCs. ATG5 expression was significantly attenuated by SR9009 treatment in mouse GCs. Conversely, Nr1d1 knockdown increased ATG5 expression in mouse GCs. Decreased NR1D1 expression at both the mRNA and protein levels was detected in the ovaries of Bmal1
-/- mice, along with elevated expression of ATG5. Dual-luciferase reporter assay and electrophoretic mobility shift assay showed that NR1D1 inhibited Atg5 transcription by binding to two putative retinoic acid-related orphan receptor response elements within the promoter. In addition, rapamycin-induced autophagy and ATG5 expression were partially reversed by SR9009 treatment in mouse GCs. Taken together, our current data demonstrated that the circadian clock regulates GC autophagy through NR1D1-mediated inhibition of ATG5 expression, and thus, plays a role in maintaining autophagy homeostasis in GCs.- Published
- 2022
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116. Multiple Strategies to Fabricate a Highly Stable 2D Cu II Cu I -Organic Framework with High Proton Conductivity.
- Author
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Zhao L, Zhu RR, Wang S, He L, Du L, and Zhao QH
- Abstract
Using multifunctional organic ligands with multiple acidic groups (carboxylate and sulfonate groups) to synthesize metal-organic frameworks (MOFs) bearing effective H-bond networks is a promising strategy to obtain highly proton conductive materials. In this work, a highly stable two-dimensional MOF, [Cu
II 5 CuI 2 (μ3 -OH)4 (H2 O)6 (L)2 (H2 L)2 ]·3H2 O (denoted as YCu161 ; H3 L = 6-sulfonaphthalene-1,4-dicarboxylic acid) containing mixed-valence [CuII 5 CuI 2 (μ3 -OH)4 ]8+ subunits, was successfully prepared. It exhibited excellent stability and temperature- and humidity-dependent proton conduction properties. Its optimal proton conductivity reached 1.84 × 10-3 S cm-1 at 90 °C and 98% relative humidity. On the basis of a crystal structure analysis, water vapor adsorption test results, and activation energy calculations, we deduced the proton conduction pathway and mechanism. Apparently, uncoordinated sulfonic and carboxyl groups and a network of abundant H-bonds inside the framework were responsible for the efficient proton transfer. Therefore, the strategy of selecting suitable bifunctional ligands to construct two-dimensional Cu-cluster-based MOFs with excellent proton conductivity is feasible.- Published
- 2021
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117. Bmal1 promotes prostaglandin E 2 synthesis by upregulating Ptgs2 transcription in response to increasing estradiol levels in day 4 pregnant mice.
- Author
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Zhao L, Yang L, Zhang J, Xiao Y, Wu M, Ma T, Wang X, Zhang L, Jiang H, Chao HW, Wang A, Jin Y, and Chen H
- Subjects
- ARNTL Transcription Factors genetics, Animals, Cells, Cultured, Cyclooxygenase 2 metabolism, Estradiol pharmacology, Female, Gestational Age, Male, Mice, Mice, 129 Strain, Mice, Inbred C57BL, Mice, Knockout, Pregnancy, Progesterone pharmacology, Transcriptional Activation drug effects, Uterus drug effects, Uterus metabolism, ARNTL Transcription Factors physiology, Cyclooxygenase 2 genetics, Dinoprostone biosynthesis, Estradiol blood
- Abstract
Prostaglandin G/H synthase 2 (PTGS2) is a rate-limiting enzyme in prostaglandin synthesis. The present study assessed the role of the uterine circadian clock on Ptgs2 transcription in response to steroid hormones during early pregnancy. We demonstrated that the core clock genes ( Bmal1 , Per2 , Nr1d1 , and Dbp ), Vegf , and Ptgs2, and their encoded proteins, have rhythmic expression in the mouse uterus from days 3.5 to 4.5 ( D3.5-4.5 ) of pregnancy. Progesterone (P
4 ) treatment of cultured uterus endometrial stromal cells (UESCs) isolated from mPer2Luciferase reporter gene knock-in mice on D4 induced a phase shift in PER2::LUCIFERASE oscillations. This P4 -induced phase shift of PER2::LUCIFERASE oscillations was significantly attenuated by the P4 antagonist RU486. Additionally, the amplitude of PER2::LUCIFERASE oscillations was increased by estradiol (E2 ) treatment in the presence of P4 . Consistently, the mRNA levels of clock genes ( Bmal1 and Per2 ), Vegf , and Ptgs2 were markedly increased by E2 treatment of UESCs in the presence of P4 . Treatment with E2 also promoted prostaglandin E2 (PGE2 ) synthesis by UESCs. Depletion of Bmal1 in UESCs by small-interfering RNA (siRNA) decreased the transcript levels of clock genes ( Nr1d1 and Dbp ), Vegf , and Ptgs2 compared with nonsilencing siRNA treatment. Bmal1 knockdown also inhibited PGE2 synthesis. Moreover, the mRNA expression levels of clock genes ( Nr1d1 and Dbp ), Vegf , and Ptgs2 , and their respective proteins were significantly decreased in the uterus of Bmal1-/- mice. Thus, these data suggest that Bmal1 in mice promotes PGE2 synthesis by upregulating Ptgs2 in response to increases in E2 on D4 of pregnancy. NEW & NOTEWORTHY Rhythmic expression of Bmal1 and Ptgs2 was observed in the uterus isolated from D3.5-4.5 of pregnant mice. E2 increased the expression of Bmal1 and Ptg2 in UESCs isolated from mice on D4. The expression of Ptgs2 was significantly decreased in Bmal1-siRNA treated UESCs. Bmal1 knockdown also inhibited PGE2 synthesis. Thus, these data suggest that Bmal1 in mice promotes PGE2 synthesis by upregulating Ptgs2 in response to increases in E2 on D4 of pregnancy.- Published
- 2021
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118. Removal of Rev-erbα inhibition contributes to the prostaglandin G/H synthase 2 expression in rat endometrial stromal cells.
- Author
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Isayama K, Zhao L, Chen H, Yamauchi N, Shigeyoshi Y, Hashimoto S, and Hattori MA
- Subjects
- 5' Untranslated Regions, ARNTL Transcription Factors antagonists & inhibitors, ARNTL Transcription Factors genetics, ARNTL Transcription Factors metabolism, Animals, Cells, Cultured, Cyclooxygenase 2 genetics, Endometrium cytology, Endometrium enzymology, Female, Nuclear Receptor Subfamily 1, Group D, Member 1 antagonists & inhibitors, Nuclear Receptor Subfamily 1, Group D, Member 1 genetics, Nuclear Receptor Subfamily 1, Group F, Member 1 genetics, Nuclear Receptor Subfamily 1, Group F, Member 1 metabolism, Pregnancy, Prolactin analogs & derivatives, Prolactin genetics, Prolactin metabolism, RNA Interference, RNA, Small Interfering, Rats, Rats, Transgenic, Response Elements, Stromal Cells cytology, Stromal Cells enzymology, Circadian Clocks, Cyclooxygenase 2 metabolism, Endometrium metabolism, Gene Expression Regulation, Enzymologic, Nuclear Receptor Subfamily 1, Group D, Member 1 metabolism, Placentation, Stromal Cells metabolism
- Abstract
The rhythmic expression of clock genes in the uterus is attenuated during decidualization. This study focused on Ptgs2, which is essential for decidualization, as a putative clock-controlled gene, and aimed to reveal the functions of clock genes in relation to Ptgs2 during decidualization. We compared the transcript levels of clock genes in the rat uterus on days 4.5 (D4.5) and 6.5 of pregnancy. The transcript levels of clock genes (Per2, Bmal1, Rorα, and Rev-erbα) had decreased at implantation sites on day 6.5 (D6.5e) compared with those on D4.5, whereas Ptgs2 transcripts had increased on D6.5e. Similar observations of Rev-erbα and Ptgs2 were also obtained in the endometrium on D6.5e by immunohistochemistry. In the decidual cells induced by medroxyprogesterone and 2-O-dibutyryl-cAMP, the rhythmic expression levels of clock genes were attenuated, whereas Ptgs2 transcription was induced. These results indicate that decidualization causes the attenuation of clock genes and the induction of Ptgs2. Furthermore, in the experiment of Bmal1 siRNA, the rhythmic expression of clock genes and Ptgs2 was attenuated by the siRNA. Transcript levels of Ptgs2 and prostaglandin (PG)E₂ production were increased by treatment with the Rev-erbα antagonist, suggesting the contribution of the nuclear receptor Rev-erbα to Ptgs2 expression. Moreover, Rev-erbα knockdown enhanced the induction of Ptgs2 transcription and PGE₂ production by forskolin. Chromatin immunoprecipitation-PCR analysis revealed that Rev-erbα could directly bind to a proximal RORE site of Ptgs2. Collectively, this study demonstrates that the attenuation of the circadian clock, especially its core component Rev-erbα, contributes to the induction of Ptgs2 during decidualization., (Copyright © 2015 the American Physiological Society.)
- Published
- 2015
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119. Inhibitory role of REV-ERBα in the expression of bone morphogenetic protein gene family in rat uterus endometrium stromal cells.
- Author
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Tasaki H, Zhao L, Isayama K, Chen H, Yamauchi N, Shigeyoshi Y, Hashimoto S, and Hattori MA
- Subjects
- Animals, Cells, Cultured, Female, Gene Expression Regulation, Pregnancy, Rats, Rats, Transgenic, Stromal Cells metabolism, Uterus cytology, Uterus metabolism, Bone Morphogenetic Proteins antagonists & inhibitors, Bone Morphogenetic Proteins biosynthesis, Endometrium cytology, Endometrium metabolism, Nuclear Receptor Subfamily 1, Group D, Member 1 biosynthesis
- Abstract
Uterus circadian rhythms have been implicated in the gestation processes of mammals through entraining of the clock proteins to numerous downstream genes. Bone morphogenetic proteins (BMPs), having clock-controlled regulatory sites in their gene promoters, are expressed in the uterus during decidualization, but the regulation of the Bmp gene expression is poorly understood. The present study was designed to dissect the physiological roles of the uterus oscillators in the Bmp expression using the uterus endometrial stromal cells (UESCs) isolated from Per2-dLuc transgenic rats on day 4.5 of gestation. The in vitro decidualization of UESCs was induced by medroxyprogesterone acetate and 2-O-dibutyryl cAMP. A significant decline of Per2-dLuc bioluminescence activity was induced in decidual cells, and concomitantly, the expression of canonical clock genes was downregulated. Conversely, the expression of the core Bmp genes Bmp2, Bmp4, Bmp6, and Bmp7 was upregulated. In UESCs transfected with Bmal1-specific siRNA, in which Rev-erbα expression was downregulated, Bmp genes, such as Bmp2, Bmp4, and Bmp6 were upregulated. However, Bmp1, Bmp7, and Bmp8a were not significantly affected by Bmal1 silencing. The expression of all Bmp genes was enhanced after treatment with the REV-ERBα antagonist (SR8278), although their rhythmic profiles were differed from each other. The binding of REV-ERBα to the proximal regions of the Bmp2 and Bmp4 promoters was revealed by chromatin immunoprecipitation-PCR analysis. Collectively, these results indicate that the Bmp genes are upregulated by the attenuation of the cellular circadian clock; in particular, its core component REV-ERBα functions as a transcriptional silencer in the Bmp gene family., (Copyright © 2015 the American Physiological Society.)
- Published
- 2015
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120. Profiling of circadian genes expressed in the uterus endometrial stromal cells of pregnant rats as revealed by DNA microarray coupled with RNA interference.
- Author
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Tasaki H, Zhao L, Isayama K, Chen H, Nobuhiko Yamauchi, Yasufumi Shigeyoshi, Hashimoto S, and Hattori MA
- Abstract
The peripheral circadian oscillator plays an essential role in synchronizing local physiology to operate in a circadian manner via regulation of the expression of clock-controlled genes. The present study aimed to evaluate the circadian rhythms of clock genes and clock-controlled genes expressed in the rat uterus endometrial stromal cells (UESCs) during the stage of implantation by a DNA microarray. Of 12,252 genes showing significantly expression, 7,235 genes displayed significant alterations. As revealed by the biological pathway analysis using the database for annotation, visualization, and integrated discovery online annotation software, genes were involved in cell cycle, glutathione metabolism, MAPK signaling pathway, fatty acid metabolism, ubiquitin mediated proteolysis, focal adhesion, and PPAR signaling pathway. The clustering of clock genes were mainly divided into four groups: the first group was Rorα, Timeless, Npas2, Bmal1, Id2, and Cry2; the second group Per1, Per2, Per3, Dec1, Tef, and Dbp; the third group Bmal2, Cry1, E4bp4, Rorβ, and Clock; the fourth group Rev-erbα. Eleven implantation-related genes and 24 placenta formation-related genes displayed significant alterations, suggesting that these genes involved in implantation and placenta formation are controlled under circadian clock. Some candidates as clock-controlled genes were evaluated by using RNA interference to Bmal1 mRNA. Down-regulation of Igf1 gene expression was observed by Bmal1 silencing, whereas the expression of Inhβa was significantly increased. During active oscillation of circadian clock, the apoptosis-related genes Fas and Caspase3 remained no significant changes, but they were significantly increased by knockdown of Bmal1 mRNA. These results indicate that clock-controlled genes are up- or down-regulated in rat UESCs during the stage of decidualization. DNA microarray analysis coupled with RNA interference will be helpful to understand the physiological roles of some oscillating genes in blastocyst implantation and placenta formation.
- Published
- 2013
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121. Downregulation of core clock gene Bmal1 attenuates expression of progesterone and prostaglandin biosynthesis-related genes in rat luteinizing granulosa cells.
- Author
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Chen H, Zhao L, Kumazawa M, Yamauchi N, Shigeyoshi Y, Hashimoto S, and Hattori MA
- Subjects
- ARNTL Transcription Factors biosynthesis, ARNTL Transcription Factors genetics, Animals, CLOCK Proteins antagonists & inhibitors, CLOCK Proteins biosynthesis, Cells, Cultured, Female, Gene Expression Regulation, Mice, Progesterone biosynthesis, Progesterone genetics, Prostaglandins biosynthesis, Rats, Rats, Transgenic, ARNTL Transcription Factors antagonists & inhibitors, CLOCK Proteins genetics, Down-Regulation genetics, Luteal Cells metabolism, Progesterone antagonists & inhibitors, Prostaglandins genetics
- Abstract
Ovarian circadian oscillators have been implicated in the reproductive processes of mammals. However, there are few reports regarding the detection of ovarian clock-controlled genes (CCGs). The present study was designed to unravel the mechanisms through which CCG ovarian circadian oscillators regulate fertility, primarily using quantitative RT-PCR and RNA interference against Bmal1 in rat granulosa cells. Mature granulosa cells were prepared from mouse Per2-destabilized luciferase (dLuc) reporter gene transgenic rats. A real-time monitoring system of Per2 promoter activity was employed to detect Per2-dLuc oscillations. The cells exposed to luteinizing hormone (LH) displayed clear Per2-dLuc oscillations and a rhythmic expression of clock genes (Bmal1, Per1, Per2, Rev-erbα, and Dbp). Meanwhile, the examined ovarian genes (Star, Cyp19a1, Cyp11a1, Ptgs2, Lhcgr, and p53) showed rhythmic transcript profiles except for Hsd3b2, indicating that these rhythmic expression genes may be CCGs. Notably, Bmal1 small interfering (si)RNA treatment significantly decreased both the amplitude of Per2-dLuc oscillations and Bmal1 mRNA levels compared with nonsilencing RNA treatment in luteinizing granulosa cells. Depletion of Bmal1 by siRNA decreased the transcript levels of clock genes (Per1, Per2, Rev-erbα, and Dbp) and examined ovarian genes (Star, Cyp19a1, Cyp11a1, Ptgs2, Hsd3b2, and Lhcgr). Accordingly, knockdown of Bmal1 also inhibited the synthesis of progesterone and prostaglandin E2, which are associated with crucial reproductive processes. Collectively, these data suggest that ovarian circadian oscillators regulate the synthesis of steroid hormones and prostaglandins through ovarian-specific CCGs in response to LH stimuli. The present study provides new insights into the physiologic significance of Bmal1 related to fertility in ovarian circadian oscillators.
- Published
- 2013
- Full Text
- View/download PDF
122. FSH induces the development of circadian clockwork in rat granulosa cells via a gap junction protein Cx43-dependent pathway.
- Author
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Chen H, Zhao L, Chu G, Kito G, Yamauchi N, Shigeyoshi Y, Hashimoto S, and Hattori MA
- Subjects
- Animals, Cell Differentiation drug effects, Cells, Cultured, Chorionic Gonadotropin metabolism, Circadian Rhythm Signaling Peptides and Proteins biosynthesis, Circadian Rhythm Signaling Peptides and Proteins genetics, Circadian Rhythm Signaling Peptides and Proteins metabolism, Connexin 43 antagonists & inhibitors, Connexin 43 genetics, Dexamethasone pharmacology, Female, GABA-A Receptor Antagonists pharmacology, Gap Junctions drug effects, Genes, Reporter drug effects, Glucocorticoids pharmacology, Granulosa Cells cytology, Granulosa Cells drug effects, Membrane Transport Modulators pharmacology, Mice, Period Circadian Proteins biosynthesis, Period Circadian Proteins genetics, Period Circadian Proteins metabolism, Rats, Rats, Transgenic, Receptors, LH biosynthesis, Receptors, LH genetics, Receptors, LH metabolism, Circadian Clocks drug effects, Connexin 43 metabolism, Follicle Stimulating Hormone metabolism, Gap Junctions metabolism, Granulosa Cells metabolism, Up-Regulation drug effects
- Abstract
The present study was designed to assess the relationship between gap junctions and the maturation of a clock system in rat granulosa cells stimulated by follicle-stimulating hormone (FSH). Immature and mature granulosa cells were prepared by puncturing the ovaries of diethylstilbestrol- and equine chorionic gonadotropin (eCG)-treated mouse Period2 (Per2)-dLuc reporter gene transgenic rats, respectively. Mature granulosa cells exposed to dexamethasone (DXM) synchronization displayed several Per2-dLuc oscillations and a rhythmic expression of clock genes. Intriguingly, we observed clear evidence that the FSH stimulation significantly increased the amplitude of Per2 oscillations in the granulosa cells, which was confirmed by the elevation of the Per2 and Rev-erbα (Nr1d1) mRNA levels. FSH also induced a major phase-advance shift of Per2 oscillations. The mature granulosa cells cultured for 2 days with FSH expressed higher mRNA levels of Per2, Rev-erbα, Bmal1 (Arnt1), Lhcgr, and connexin (Cx) 43 (Gja1) compared with the immature granulosa cells. Consistently, our immunofluorescence results revealed abundant Cx43 protein in antral follicles stimulated with eCG and weak or no fluorescence signal of Cx43 in primary and preantral follicles. Similar results were confirmed by Western blotting analysis. Two gap junction blockers, lindane and carbenoxolone (CBX), significantly decreased the amplitude of Per2 oscillations, which further adhered significant decreases in Per2 and Rev-erbα transcript levels. In addition, both lindane and CBX induced a clear phase-delay shift of Per2 oscillations. These findings suggest that FSH induces the development of the clock system by increasing the expression of Cx43.
- Published
- 2013
- Full Text
- View/download PDF
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