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1.
J Cell Physiol ; 239(6): e31289, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38685566

RESUMO

Follicular cysts are a common reproductive disorder in domestic animals that cause considerable economic losses to the farming industry. Effective prevention and treatment methods are lacking because neither the pathogenesis nor formation mechanisms of follicular cysts are well-understood. In this study, we first investigated the granulosa cells (GCs) of cystic follicles isolated from pigs. We observed a significant reduction in the expression of methyltransferase-like 3 (METTL3). Subsequent experiments revealed that METTL3 downregulation in GCs caused a decrease in m6A modification of pri-miR-21. This reduction further inhibited DGCR8 recognition and binding to pri-miR-21, dampening the synthesis of mature miR-21-5p. Additionally, the decrease in miR-21-5p promotes IL-1ß expression in GCs. Elevated IL-1ß activates the NFκB pathway, in turn upregulating apoptotic genes TNFa and BAX/BCL2. The subsequent apoptosis of GCs and inhibition of autophagy causes downregulation of CYP19A1 expression. These processes lower oestrogen secretion and contribute to follicular cyst formation. In conclusion, our findings provide a foundation for understanding and further exploring the mechanisms of follicular-cyst development in farm animals. This work has important implications for treating ovarian disorders in livestock and could potentially be extended to humans.


Assuntos
Apoptose , Células da Granulosa , Metiltransferases , MicroRNAs , Animais , Feminino , Apoptose/genética , Células Cultivadas , Regulação para Baixo , Cisto Folicular/genética , Cisto Folicular/patologia , Cisto Folicular/metabolismo , Células da Granulosa/metabolismo , Interleucina-1beta/metabolismo , Interleucina-1beta/genética , Metiltransferases/metabolismo , Metiltransferases/genética , MicroRNAs/genética , MicroRNAs/metabolismo , NF-kappa B/metabolismo , NF-kappa B/genética , Transdução de Sinais , Suínos , Proteínas de Ligação a RNA/metabolismo
2.
Biol Reprod ; 2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-38938081

RESUMO

Ovarian theca cells produce testosterone, which acts as a vital precursor substance for synthesizing estrogens during follicular development. Nerve growth factor (NGF) has been shown to participate in reproductive physiology, specifically to follicular development and ovulation. There is currently no available data on the impact of NGF on testosterone synthesis in porcine theca cells. Furthermore, m6A modification is the most common internal modification in eukaryotic mRNAs that are closely associated with female gametogenesis, follicle development, ovulation, and other related processes. It is also uncertain whether the three main enzymes associated with m6A, such as Writers, Erasers and Readers, play a role in this process. The present study, with an in vitro culture model, investigated the effect of NGF on testosterone synthesis in porcine theca cells and the role of Writers-METTL14 in this process. It was found that NGF activates the PI3K/AKT signaling pathway through METTL14, which regulates testosterone synthesis in porcine theca cells. This study will help to further elucidate the mechanisms by which NGF regulates follicular development and provide new therapeutic targets for ovary-related diseases in female animals.

3.
Int J Mol Sci ; 25(4)2024 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-38397033

RESUMO

In female mammals, the proliferation and apoptosis of granulosa cells (GCs) are critical in determining the fate of follicles and are influenced by various factors, including brain-derived neurotrophic factor (BDNF). Previous research has shown that BDNF primarily regulates GC proliferation through the PI3K/AKT, NF-kB, and CREB tumour pathways; however, the role of other molecular mechanisms in mediating BDNF-induced GC proliferation remains unclear. In this study, we investigated the involvement of the m6A reader YTH domain-containing family member 2 (YTHDF2) in BDNF-stimulated GC proliferation and its underlying mechanism. GCs were cultured in DMEM medium supplemented with varying BDNF concentrations (0, 10, 30, 75, and 150 ng/mL) for 24 h. The viability, number, and cell cycle of GCs were assessed using the CCK-8 assay, cell counting, and flow cytometry, respectively. Further exploration into YTHDF2's role in BDNF-stimulated GC proliferation was conducted using RT-qPCR, Western blotting, and sequencing. Our findings indicate that YTHDF2 mediates the effect of BDNF on GC proliferation. Additionally, this study suggests for the first time that BDNF promotes YTHDF2 expression by increasing the phosphorylation level of the ERK1/2 signalling pathway. This study offers a new perspective and foundation for further elucidating the mechanism by which BDNF regulates GC proliferation.


Assuntos
Fator Neurotrófico Derivado do Encéfalo , Fosfatidilinositol 3-Quinases , Feminino , Suínos , Animais , Fator Neurotrófico Derivado do Encéfalo/genética , Fator Neurotrófico Derivado do Encéfalo/farmacologia , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Células da Granulosa/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismo , Proliferação de Células , Mamíferos/metabolismo
4.
Small ; 19(25): e2208101, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36932880

RESUMO

Multifunctional applications including efficient microwave absorption and electromagnetic interference (EMI) shielding as well as excellent Li-ion storage are rarely achieved in a single material. Herein, a multifunctional nanocrystalline-assembled porous hierarchical NiO@NiFe2 O4 /reduced graphene oxide (rGO) heterostructure integrating microwave absorption, EMI shielding, and Li-ion storage functions is fabricated and tailored to develop high-performance energy conversion and storage devices. Owing to its structural and compositional advantages, the optimized NiO@NiFe2 O4 /15rGO achieves a minimum reflection loss of -55 dB with a matching thickness of 2.3 mm, and the effective absorption bandwidth is up to 6.4 GHz. The EMI shielding effectiveness reaches 8.69 dB. NiO@NiFe2 O4 /15rGO exhibits a high initial discharge specific capacity of 1813.92 mAh g-1 , which reaches 1218.6 mAh g-1 after 289 cycles and remains at 784.32 mAh g-1 after 500 cycles at 0.1 A g-1 . In addition, NiO@NiFe2 O4 /15rGO demonstrates a long cycling stability at high current densities. This study provides an insight into the design of advanced multifunctional materials and devices and provides an innovative method of solving current environmental and energy problems.

5.
Small ; 19(43): e2303186, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37365955

RESUMO

With the advent of intelligent society and the popularity of electronic equipment, the protection and treatment of electromagnetic (EM) radiation have become hot research topics all over the world. Herein, novel 2D carbon-based nanoplates with uniformly embedded Co nanoparticles are prepared, with unique hierarchical structure and integrated magnetic-dielectric components. The obtained hierarchical nanoplates exhibit a wide range of tunable EM properties (ε' for 3.38 to 34.67 and ε″ for 0.13 to 31.45) by manipulating the dispersed states inside wax system, which can achieve an effective switch from microwave absorption to EM interference shielding performance. The optimal reflection loss reaches -55.6 dB, and the shielding efficiency is 93.5%. Meanwhile, the hierarchical nanoplates also exhibit impressive capacitive performance, with a specific capacitance of 1654 F g-1 at 1 A g-1 . Based on this, a creative device is constructed with the nanoplates, which can convert harmful EM radiation to useful electric energy for recycling. This work offers a new idea for the development of EM materials and functional devices, powerfully promoting the advance of energy and environmental fields.

6.
Small ; 18(51): e2205624, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36328711

RESUMO

Electrical conductivity and dielectric parameters are general inherent features of materials. Controlling these characteristics through applied bias will add a new dimension to regulate the dynamic response of smart materials. Here, a fascinating electrical transport behavior is observed in topological insulator (TI) Bi2 Te3 nanorods, which will play a vital role in intelligent materials or devices as a unit for information reception, processing or feedback. The Bi2 Te3 nanorod aggregates exhibit a monotonic resistance response to voltage, with observed four-fold change of electrical conductivity in a small range electric field of 1 V mm-1 . The dielectric constant and dielectric loss of Bi2 Te3 nanorod composites also show strong dependences on bias voltage due to the unique electrical transport characteristics. The unique voltage-controlled electrical responses are attributed to the change of Fermi levels within the band structure of disordered TI nanorods, which are non-parallel to the applied electric field. The excellent controllable inherent characteristics through electric field endows Bi2 Te3 nanomaterials bright prospects for applications in smart devices and resistive random access memories.

7.
FASEB J ; 35(5): e21610, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33908671

RESUMO

Granulosa cells (GCs) are regulated by various factors during ovarian development. However, there are few reports on the role of follicular fluid exosomes in ovarian GCs. In this study, porcine ovarian GCs were used to explore the effects of follicular fluid exosomes on GCs. GCs were treated with in vitro, and the changes in cell proliferation, steroid synthesis, and associated signal pathways were detected. The results showed that exosomes increased cell viability and altered the gene expression profile of GCs. Exosomes also increased the level of gene expression associated with both proliferation and progesterone synthesis, in which the MAPK/ERK and WNT/B-CATENIN pathways were involved. In addition, exosome-carried microRNAs were identified by high-throughput sequencing, and exosomal miR-31-5p was found to promote the proliferation of GCs and progesterone synthesis via the WNT/B-CATENIN pathway by targeting the SFRP4 follicle growth inhibitor. In conclusion, this study has demonstrated that exosomes are essential substances involved in regulating the physiological function of GCs.


Assuntos
Proliferação de Células , Exossomos/metabolismo , Líquido Folicular/metabolismo , Células da Granulosa/citologia , MicroRNAs/genética , Folículo Ovariano/citologia , Esteroides/biossíntese , Animais , Apoptose , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Células da Granulosa/metabolismo , Folículo Ovariano/metabolismo , Suínos
8.
Nanotechnology ; 33(29)2022 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-35381578

RESUMO

Micro-supercapacitors (MSCs) are considered as the promising energy supply of miniaturized electronic devices. The electrode material, as one integral part, play a crucial role on the energy storage performance of MSCs. In our work, we constructed a heterojunction in MXene-PANI@MWCNTs (MPM) ternary composite, benefitting for the synergistic enhancement effect among MXene, polyaniline (PANI) and multiwall carbon nanotubes, an outstanding specific capacitance of 414 F g-1(at 1 A g-1) has been achieved. MPM shows high capacitance retention at large current density (86.7%, at 10 A g-1) and long-term cycling stability of 90.4% for 10 000 cycles. Furthermore, we obtained MPM self-standing films, and constructed a flexible all-solid-state MSC based on the film electrode. A competitive charge storage capability of 30.2 mF cm-2and long-term stability of 70.2% retention for 10 000 cycles was obtained in the MSC. Meanwhile, the MSC shows excellent flexibility, maintaining most capacitance under bending conditions. Moreover, using an integrated strategy, MSCs can obtain tunable voltages and currents that meet various practical requirements. All these results indicate that the MPM is an excellent charge storage material and will become a potential candidate for flexible energy-storage devices.

9.
Nanotechnology ; 32(44)2021 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-34320474

RESUMO

Ti3C2Txis an important member of the MXenes family. Due to its excellent electrical conductivity, adjustable atomic layer, and modifiable active surface, Ti3C2Txhas attracted great attention in the field of electromagnetic interference (EMI) shielding. This paper introduces the important role of regulating conductive network to improve the EMI shielding performance of materials and summarizes the EMI shielding performance of Ti3C2Txnanohybrids reported in recent years. In addition, Ti3C2Txbased EMI shielding materials towards multifunctional devices are also systematically introduced. After that, the development status of Ti3C2Txnanohybrids in the field of EMI shielding is objectively described, and the main problems and challenges are evaluated. Finally, the prospect of Ti3C2Txnanohybrids for advanced and green EMI shielding materials is forecasted.

10.
Reproduction ; 160(3): 405-415, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32567558

RESUMO

Nerve growth factor (NGF) has been proved to play important roles in male reproductive physiology, but the molecular mechanisms of NGF action remain unclear. In this study, the effects of NGF on the growth of newborn bovine testicular Sertoli (NBS) cells and the related signaling pathways were investigated. The NBS cells were treated in vitro with NGF (100 ng/mL) for 18 h. The expression levels of cell proliferation related genes, INHBB, and cytoplasmic specialization related gene were determined using real-time PCR and Western blot. The roles of PI3K/AKT and MAPK/ERK pathways in NGF-induced cell proliferation were investigated. It was found that NGF regulates proliferation and function of NBS cells via its receptor NTRK1 by activating the PI3K/ATK and MAPK/ERK signaling pathways. The study will help to further understand the role of NGF in male reproduction and provide new therapeutic targets for reproductive dysfunctions in male animals.


Assuntos
Proliferação de Células , Regulação da Expressão Gênica/efeitos dos fármacos , Fator de Crescimento Neural/farmacologia , Células de Sertoli/citologia , Testículo/citologia , Animais , Animais Recém-Nascidos , Bovinos , Sistema de Sinalização das MAP Quinases , Masculino , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor trkA/genética , Receptor trkA/metabolismo , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/metabolismo , Transdução de Sinais , Testículo/efeitos dos fármacos , Testículo/metabolismo
11.
Soft Matter ; 16(31): 7332-7341, 2020 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-32685953

RESUMO

We present a method for actuating LCE materials by microwave radiation. The microwave actuation performance of a polysiloxane-based nematic liquid crystalline elastomer (LCE) was investigated. The microwave-material interaction caused a dipolar loss, which created a heating effect to trigger the nematic-isotropic transition of the LCE matrix, thus leading to the deformation actuation of the LCE material. This energy conversion from radiant energy to thermal energy provided a contactless pathway to actuate the LCE material without the aid of other components acting as energy converters. The LCE demonstrated rapid maximum contraction upon microwave irradiation, and this microwave-stimulated response was fully reversible when the microwave irradiation was switched off. More importantly, the microwave actuation exhibited superiority relative to photo-actuation, which is the usual method of contactless actuation. The microwaves can penetrate the opaque thick barriers to effectively actuate the LCE due to their strong penetrability; they can also penetrate multiple LCE samples and actuate them almost simultaneously. By taking advantage of the salient features of microwave actuation, a microwave detector system, implementing the LCE as an actuator material, was fabricated. This demonstrated the performance of monitoring microwave irradiation intensities with good sensitivity and convenient manipulation.

12.
Nanotechnology ; 31(32): 325703, 2020 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-32315995

RESUMO

Electromagnetic (EM) wave absorbing materials have been a research hotspot in materials science and related technical fields in recent years. Finding lightweight, efficient, and broadband electromagnetic wave absorbing materials has always been a very challenging subject. Herein, we successfully prepared Co3O4-WS2 hybrid nanosheets with heterostructures, which excellent combine the advantages of magnetic property of Co3O4 and dielectric property of WS2. By the electromagnetic synergy effect, maximum RL is up to -61.1 dB at 1.9 mm, and the bandwidth exceeds 5 GHz. Such highly efficient microwave absorption is attributed to not only the electromagnetic synergy effect, but the dipole polarization as well as conduction loss. These results show that the obtained Co3O4-WS2 is an excellent EM wave absorbing material and can be used as a candidate for advanced EM wave absorbing materials in the fields such as commerce, military and aerospace in future.

13.
Lasers Med Sci ; 35(6): 1349-1355, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31897814

RESUMO

While zona pellucida (ZP) breaching of day-3 frozen blastocysts embryos can increase the blastocyst hatching rate, compared with ZP thinning, the pregnancy and implantation rates are similar. The aim of this study was to compare pregnancy outcomes and the risks associated with frozen-thawed blastocysts between laser ZP breaching and laser ZP thinning. For the thinning group, ZP of thawed blastocyst was thinned to a length of 30-40 µm using laser between January 2013 and October 2015. On the other hand, for the breaching group, thawed blastocysts were breached with a 60-80 µm hole in the ZP using laser between November 2015 and April 2018. The implantation rate of ZP breaching (72.7%) was higher than that of ZP thinning (61.8%). In single frozen blastocyst transfer, the implantation rate, clinical pregnancy rate, and live birth rate of ZP breaching (73.9%, 73.9%, 61.8%, respectively) were significantly higher than those of ZP thinning (60.9%, 60.9%, 46.7%, respectively). The abortion rate, preterm birth rate, congenital malformation, birth defects, and birth weight did not significantly differ between the two groups. In conclusion, laser assisted hatching during single frozen blastocyst transfer using ZP breaching exhibit higher implantation, pregnancy, and live birth rates compared with ZP thinning. No significant differences were observed between the two assisted hatching methods in terms of adverse effects on pregnancy and newborns.


Assuntos
Coeficiente de Natalidade , Implantação do Embrião , Transferência Embrionária , Congelamento , Lasers , Zona Pelúcida/efeitos da radiação , Adulto , Transferência Embrionária/métodos , Feminino , Fertilização in vitro , Humanos , Recém-Nascido , Luz , Masculino , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Fatores de Risco
14.
J Cell Physiol ; 234(9): 15856-15871, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30770557

RESUMO

The p75 neurotrophin receptor (p75NTR), a member of tumor necrosis factor receptor superfamily, involves in neuronal apoptosis after intracerebral hemorrhage (ICH). It has been previously demonstrated that phosphorylation of p35 is a crucial factor for fighting against the proapoptotic p25/CDK5 signaling in neuronal apoptosis. Then, in ICH models of rats and primary cortical neurons, we found that the expressions of p75NTR, p-histone H1 (the kinase activity of CDK5), p25, Fas-associated phosphatase-1 (FAP-1), and phosphorylated myocyte enhancer factor 2D (p-MEF2D) were enhanced after ICH, whereas the expression of p35-Thr(138) was attenuated. Coimmunoprecipitation analysis indicated several interactions as follows: p35/p25 and CKD5, p75NTR and p35, as well as p75NTR and FAP-1. After p75NTR or FAP-1 depletion with double-stranded RNA interference in PC12 cells, the levels of p25 and p-histone H1 were attenuated, whereas p35-Thr(138) was elevated. Considering p75NTR has no effect of dephosphorylation, our results suggested that p75NTR might promote the dephosphorylation of p35-Thr(138) via interaction with FAP-1, and the p75NTR/p35 complex upregulated p25/CDK5 signaling to facilitate the neuronal apoptosis following ICH. So, in the study, we aimed to provide a theoretical and experimental basis that p75NTR could be regulated to reduce neuronal apoptosis following ICH for potential clinical treatment.

15.
J Cell Physiol ; 234(8): 14058-14067, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30628054

RESUMO

The brain-derived neurotrophic factor (BDNF) was first recognized for its roles in the peripheral and central nervous systems, and its complex functions on mammalian organs have been extended constantly. However, to date, little is known about its effects on the male reproductive system, including the steroidogenesis of mammals. The purpose of this study was to elucidate the effects of BDNF on testosterone generation of Leydig cells and the underlying mechanisms. We found that BDNF-induced proliferation of TM3 Leydig cells via upregulation of proliferating cell nuclear antigen ( Pcna) and promoted testosterone generation as a result of upregulation of steroidogenic acute regulatory protein ( Star), 3b-hydroxysteroid dehydrogenase ( Hsd3b1), and cytochrome P450 side-chain cleavage enzyme ( Cyp11a1) both in primary Leydig cells and TM3 Leydig cells, which were all attenuated in Bdnf knockdown TM3 Leydig cells. Furthermore, the possible mechanism of testosterone synthesis was explored in TM3 Leydig cells. The results showed that BDNF enhanced extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) phosphorylation, and the effect was disrupted by Bdnf deletion. Moreover, PD98059, a potent selective inhibitor of ERK1/2 activation, compromised BDNF-induced testosterone generation and upregulation of Star, Hsd3b1, and Cyp11a1. The Bdnf knockdown assay, on the other hand, indicated the autocrine effect of BDNF on steroidogenesis in TM3 Leydig cells. On the basis of these results, we concluded that BDNF, acting as an autocrine factor, induced testosterone generation as a result of the upregulation of Star, Hsd3b1, and Cyp11a1 via stimulation of the ERK1/2 pathway.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/genética , Fosfoproteínas/genética , Antígeno Nuclear de Célula em Proliferação/genética , Reprodução/genética , Testosterona/biossíntese , Animais , Comunicação Autócrina/genética , Fator Neurotrófico Derivado do Encéfalo/farmacologia , Proliferação de Células/efeitos dos fármacos , Enzima de Clivagem da Cadeia Lateral do Colesterol/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Técnicas de Silenciamento de Genes , Células Intersticiais do Testículo/efeitos dos fármacos , Células Intersticiais do Testículo/metabolismo , Sistema de Sinalização das MAP Quinases/genética , Masculino , Camundongos , Complexos Multienzimáticos/genética , Progesterona Redutase/genética , Esteroide Isomerases/genética , Testosterona/genética
16.
Soft Matter ; 15(30): 6116-6126, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31286128

RESUMO

In this work, according to the characteristic of surface plasmon resonance (SPR) of metallic nanoparticles, we investigated the photo actuation performance of a liquid crystalline elastomer (LCE) nanocomposite with incorporated gold nanoparticles (nano-gold/LCE nanocomposite). The nano-gold/LCE nanocomposites were fabricated by incorporating gold nanoparticles into a polysiloxane-based LCE matrix via a novel experimental protocol, and characterized by a well-developed SPR absorption band in the visible spectrum range. The nano-gold/LCE nanocomposites demonstrated strong actuation upon irradiation with a quasi-daylight source; the reason lay in that the SPR response of gold nanoparticles performed efficient energy conversion from light energy to thermal energy, and thus offered an activation pathway for the nematic-isotropic transition of the LCE matrix. The nano-gold/LCE nanocomposites underwent rapid maximum axial contraction up to about one third of the original length under light irradiation, and this photo-stimulated muscle-like actuation was fully reversible via the on-off switching of the light source. The photo actuation properties of nano-gold/LCE nanocomposites with varying irradiation intensities and gold nanoparticle content were also investigated. In addition, the nano-gold/LCE nanocomposites demonstrated superior optical nonlinear properties, and revealed potential for the application area of mode-locking for laser technology.

17.
Nanotechnology ; 30(44): 445708, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31349245

RESUMO

Rational structure design of microwave absorption material is extremely significant from the perspectives of enhancing the electromagnetic microwave absorption (EMA) performance and adapting to cost-effective and sustainable industrial applications. Here, reduced graphene oxides (rGOs) with curl structures derived from corn stover are applied for the absorption of electromagnetic waves. The results suggest that biomass-rGO show the maximum reflection loss of -51.7 dB and an effective absorption bandwidth 13.5 GHz (4.5-18 GHz) at a thickness of 3.25 mm, implying the unique critical role of the microstructure in adjusting the EMA performance. Moreover, the successful conversion of waste biomass into widely used electromagnetic wave absorbing materials could solve the problems of environmental pollution caused by straw burning.

18.
Biotechnol Lett ; 41(8-9): 915-920, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31256272

RESUMO

OBJECTIVES: To develop an efficient, economical, and low-toxicity method for the extraction of RNA from animal cells to meet a basic requirement of biological research: the isolation of high-quality RNA. RESULTS: Guanidine hydrochloride was used as a lysis buffer and Na-acetate was used as a wash buffer to extract RNA fragments from TM3 Leydig cells and ovarian granulosa cells efficiently. The functionality of the extracted RNA samples was verified through polymerase chain reaction (PCR) and real-time fluorescence quantitative PCR (RT-PCR). PCR results showed that the normal DNA column-based method could guarantee RNA integrity and could be used to amplify gene fragments successfully. RT-PCR analysis showed that the RNA samples isolated through the proposed method could be used to detect the expression levels of steroidogenic acute regulatory protein and hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 mRNA in TM3 Leydig cells under induction by luteinizing hormone. The proposed method could be used to isolate RNA from mammalian cells and provided RNA yields of > 120 ng/5 × 106 cells. This method provided RNA with purities and yields that are sufficient for cDNA synthesis and PCR amplification in gene expression studies. CONCLUSIONS: The proposed RNA extraction method has the advantages of low toxicity, safe handling, and low cost. Isolation can be completed in 20 min. The proposed method can be used to extract RNA from various animal cell samples and is worth promoting.


Assuntos
Células da Granulosa/química , Células Intersticiais do Testículo/química , Biologia Molecular/métodos , RNA/isolamento & purificação , Animais , Feminino , Masculino , Mamíferos , Reação em Cadeia da Polimerase , RNA/genética
19.
Int J Mol Sci ; 20(3)2019 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-30704086

RESUMO

Ammonia is one of the major toxic components of metabolites in blood and tissues of high-producing dairy cows and could affect the health of bovine mammary glands. Bovine mammary epithelial cells are sensitive to oxidative stress induced by intensive cell metabolism. In our previous study, we found that ammonia could induce oxidative stress, apoptosis and inflammatory responses in bovine mammary epithelial cells. In the present study, the cytoprotective effects of astragaloside IV against ammonia in vitro were explored. The results demonstrated that pretreatment of MAC-T cells with astragaloside IV could potently suppress the increase in the level of intracellular reactive oxygen species (ROS) and the rate of cell apoptosis, inhibit the ammonia-induced inflammatory responses, and rescue the decrease of cell viability. Astragaloside IV prevented ammonia-induced endoplasmic reticulum stress. Astragaloside IV also significantly suppressed the levels of BAX, caspase 3 and p53 phosphorylation in ammonia-induced MAC-T cells. Nuclear factor erythroid 2-related factor 2(Nrf2) was essential for cytoprotective effects of astragaloside IV in MAC-T cells, as knockdown of Nrf2 dramatically abolished the prosurvival effects of astragaloside IV on treated cells. Furthermore, the PI3K/AKT and ERK/MAPK pathways were responsible for the induction of Nrf2 by astragaloside IV. In conclusion, astragaloside IV played a beneficial role against ammonia-induced damage of MAC-T cells. This provides a cue for future study to use astragaloside IV as a protective and curative agent against ammonia exposure of mammary glands in dairy cows.


Assuntos
Amônia/farmacologia , Apoptose/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Glândulas Mamárias Humanas/citologia , Estresse Oxidativo/efeitos dos fármacos , Animais , Bovinos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Feminino , Humanos , Fosforilação/efeitos dos fármacos , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Saponinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Triterpenos/farmacologia
20.
Small ; : e1800987, 2018 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-29882284

RESUMO

Electromagnetic energy radiation is becoming a "health-killer" of living bodies, especially around industrial transformer substation and electricity pylon. Harvesting, converting, and storing waste energy for recycling are considered the ideal ways to control electromagnetic radiation. However, heat-generation and temperature-rising with performance degradation remain big problems. Herein, graphene-silica xerogel is dissected hierarchically from functions to "genes," thermally driven relaxation and charge transport, experimentally and theoretically, demonstrating a competitive synergy on energy conversion. A generic approach of "material genes sequencing" is proposed, tactfully transforming the negative effects of heat energy to superiority for switching self-powered and self-circulated electromagnetic devices, beneficial for waste energy harvesting, conversion, and storage. Graphene networks with "well-sequencing genes" (w = Pc /Pp > 0.2) can serve as nanogenerators, thermally promoting electromagnetic wave absorption by 250%, with broadened bandwidth covering the whole investigated frequency. This finding of nonionic energy conversion opens up an unexpected horizon for converting, storing, and reusing waste electromagnetic energy, providing the most promising way for governing electromagnetic pollution with self-powered and self-circulated electromagnetic devices.

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