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1.
J Steroid Biochem Mol Biol ; 198: 105537, 2019 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-31785377

RESUMO

Porcine pancreatic stem cells (pPSCs) can be induced to insulin-secreting cells and therefore considered the most promising seeding cells for curing human diabetes in future. However, insufficient pPSCs number is one of the bottleneck problems before its clinical application. SerpinB1 is a serine protease inhibitor in neutrophils and can directly promote the proliferation of ß cells. Whether SerpinB1 is involved in pPSC proliferation and differentiation remains unknown. The effects of SerpinB1 on pPSCs proliferation were measured by Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine, qRT-PCR, western blot, and flow cytometry assays. We found that pPSCs did not efficiently reach the S phase when SerpinB1 expression was knocked down with short hairpin RNA (sh-SerpinB1), the expression of Cyclin D1, CDK-2, and PCNA also decreased. Meanwhile, cell viability and proliferation ability were both declined. Further analyses showed that the expression level of phosphorylated STAT3/STAT3was downregulated, along with an upregulation of p53 and p21. We used a two-step induction method to induce pPSCs to insulin-secreting cells and found that SerpinB1 expression in insulin-secreting cells was higher than in pPSCs. Meanwhile, the protein expression level of phosphorylated STAT3/STAT3 was increased while p53 and p21 was decreased in induced insulin-secreting cells in comparison with control cells. The insulin-secreting cells derived from the sh-SerpinB1 cells secreted less insulin and showed poor sensitivity to high glucose than control group. However, the insulin-secreting cells derived from the ov-SerpinB1 cells has a quite contrary tendency. In conclusion, this study demonstrates that SerpinB1 promotes the proliferation of pPSCs through the STAT3 signaling pathway, and SerpinB1 is a key factor for maintaining the viability of pPSCs during the transition to insulin-secreting cells.

2.
Res Vet Sci ; 126: 233-239, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31635840

RESUMO

In this study, canine adipose-derived mesenchymal stem cells (cADSCs) therapeutic potential was investigated in artificially induced acute liver injury model by CCl4 in canines. The primary cADSCs cells were cultured and then intravenously administered into the canine animal model. Six cross-breed dogs were divided into three groups including blank control group, CCl4 model group, CCl4 induced cADSCs transplantation group. The results showed that after intraperitoneal injection of CCl4 solution, the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and Albumin (ALB) in peripheral blood of experimental canines confirmed the correct induction of acute liver injury. Moreover, the liver structure showed clear macroscopic damage. The cADSCs were homed in the liver of the administered animals. The AST, ALT and ALB in the peripheral blood rapidly decreased. H&E and PAS histological evaluation showed that both the structure of canine liver tissue and the ability to synthesize hepatic glycogen could be restored to the control level after cADSCs transplantation. Therefore, cADSCs can play a therapeutic role in the recovery of liver injury. Overall, this study demonstrates that the primary cADSCs transplantation into the acute liver injury model induced by intravenous injection can play a certain therapeutic role in the recovery of liver in canines. These results may provide a new treatment idea for acute liver disease in pets clinically.

3.
Environ Sci Pollut Res Int ; 26(28): 28817-28828, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31377928

RESUMO

A review of energy subsidy research from a bibliometric perspective was conducted. Based on the bibliometric method, a statistical analysis of energy subsidy-related publications from 1997 to 2016 was undertaken using the Science Citation Index (SCI) and Social Science Citation Index (SSCI) databases. A total of 1182 publications were retrieved, with a significant increase in the number of publications observed after 2006. The majority of these publications were within the disciplines of Energy & Fuels and Environmental Science & Ecology. Although the USA and China contributed the most papers, authors from 96 countries were involved in the various studies. The USA was the center of global collaborations, while other countries/territories mainly conducted bilateral or regional collaborations in their research activities. Five of the top 11 most productive institutes were from China, followed by the USA. The frequency of collaborations among institutes was relatively low. However, the institute-keyword 2-mode network showed that institutes had great potential to cooperate on a number of common topics. Five major themes were identified from the co-keywords analysis: general renewable energy research, bio-energies, sustainability, subsidies, and welfare. The findings, as a complement to previous conventional reviews, will be useful in future energy subsidy research.

4.
Cell Prolif ; 52(3): e12591, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30896067

RESUMO

OBJECTIVES: To date, many efforts have been made to establish porcine embryonic stem (pES) cells without success. Extraembryonic endoderm (XEN) cells can self-renew and differentiate into the visceral endoderm and parietal endoderm. XEN cells are derived from the primitive endoderm of the inner cell mass of blastocysts and may be an intermediate state in cell reprogramming. MATERIALS AND METHODS: Porcine XEN cells (pXENCs) were generated from porcine pluripotent stem cells (pPSCs) and were characterized by RNA sequencing and immunofluorescence analyses. The developmental potential of pXENCs was investigated in chimeric mouse embryos. RESULTS: Porcine XEN cells derived from porcine pPSCs were successfully expanded in N2B27 medium supplemented with bFGF for least 30 passages. RNA sequencing and immunofluorescence analyses showed that pXENCs expressed the murine and canine XEN markers Gata6, Gata4, Sox17 and Pdgfra but not the pluripotent markers Oct4, Sox2 and TE marker Cdx2. Moreover, these cells contributed to the XEN when injected into four-cell stage mouse embryos. Supplementation with Chir99021 and SB431542 promoted the pluripotency of the pXENCs. CONCLUSIONS: We successfully derived pXENCs and showed that supplementation with Chir99021 and SB431542 confer them with pluripotency. Our results provide a new resource for investigating the reprogramming mechanism of porcine-induced pluripotent stem cells.


Assuntos
Endoderma/citologia , Endoderma/embriologia , Suínos/embriologia , Animais , Técnicas de Cultura de Células , Diferenciação Celular , Linhagem Celular , Técnicas de Cocultura , Cães , Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/metabolismo , Endoderma/metabolismo , Expressão Gênica , Camundongos , Células-Tronco Pluripotentes/citologia , Células-Tronco Pluripotentes/metabolismo , Análise de Sequência de RNA , Transdução de Sinais , Suínos/genética , Suínos/metabolismo , Quimeras de Transplante
5.
Int J Mol Sci ; 20(4)2019 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-30781644

RESUMO

Oxidative stress is the main pathogenesis of diabetic microangiopathy, which can cause microvascular endothelial cell damage and destroy vascular barrier. In this study, it is found that carnosol protects human microvascular endothelial cells (HMVEC) through antioxidative mechanisms. First, we measured the antioxidant activity of carnosol. We showed that carnosol pretreatment suppressed tert-butyl hydroperoxide (t-BHP)-induced cell viability, affected the production of lactate dehydrogenase (LDH) as well as reactive oxygen species (ROS), and increased the produce of nitric oxide (NO). Additionally, carnosol promotes the protein expression of vascular endothelial cadherin (VE-cadherin) to keep the integrity of intercellular junctions, which indicated that it protected microvascular barrier in oxidative stress. Meanwhile, we investigated that carnosol can interrupt Nrf2-Keap1 protein-protein interaction and stimulated antioxidant-responsive element (ARE)-driven luciferase activity in vitro. Mechanistically, we showed that carnosol promotes the expression of heme oxygenase 1(HO-1) and nuclear factor-erythroid 2 related factor 2(Nrf2). It can also promote the expression of endothelial nitric oxide synthase (eNOS). Collectively, our data support the notion that carnosol is a protective agent in HMVECs and has the potential for therapeutic use in the treatments of microvascular endothelial cell injury.


Assuntos
/farmacologia , Antioxidantes/farmacologia , Células Endoteliais/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , /química , Antígenos CD/metabolismo , Benzotiazóis/metabolismo , Caderinas/metabolismo , Linhagem Celular , Citoproteção/efeitos dos fármacos , Citoproteção/genética , Células Endoteliais/efeitos dos fármacos , Depuradores de Radicais Livres/metabolismo , Humanos , Microvasos/patologia , Simulação de Acoplamento Molecular , Óxido Nítrico/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Ácidos Sulfônicos/metabolismo , terc-Butil Hidroperóxido
6.
Sci Total Environ ; 663: 198-205, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30711586

RESUMO

Domestic trade flourishes with economic development and the spatial separation of production and consumption. Therefore, the prosperity of trade is accompanied by the transfer of pollution from the demand side to the supply side, which could potentially worsen the environmental quality of the supply side. Despite a large number of studies on the pollution haven hypothesis in international trade, little attention has been paid to testing the hypothesis in domestic trade. Here, combining a multiregional input-output analysis and a gravity model of trade in China, we provide an empirical test to address this problem for the first time. We also assess the factors affecting the SO2 emissions embodied in trade, including population, economic development, coal consumption, distance, and environmental regulations. We found that domestic trade contributed approximate one third of the total SO2 emissions in China, and interprovincial transfers of SO2 embodied in trade were significantly determined by the population, economic development, coal consumption of the trade pairs, as well as their distance. SO2 emission mitigation policies, such as emission reduction target and sulfur dioxide control zone, has a more significant influence on the direct transfer of SO2 emission via direct bilateral trade, while their effects were largely offset by indirect trade (through third-party transfers). Our results do not support the pollution haven hypothesis existed in domestic trade in China during 2007-2012. Our paper sets an example and provides a reference for the domestic pollution transfer problem from an econometric perspective. Further attempts on testing pollution haven hypothesis in consideration of various pollutants are still needed to arrive at a robust conclusion.

7.
J Cell Physiol ; 234(6): 8113-8121, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30317605

RESUMO

LIN28A serves as a crucial marker of dairy goat male germline stem cells (GmGSCs). In our previous study, we demonstrated that LIN28A promotes proliferation, self-renewal, and maintains the stemness of GmGSCs. Here, we found that LIN28A could activate the transcription of NANOG in a let-7g independent manner. We cloned the 5' upstream of two NANOG genes which were located on chromosome 15 ( NANOG-ch15) and chromosome 5 ( NANOG-ch5), respectively, and then examined their promoter activities and promoter methylation levels. Results showed that NANOG-ch15 is a pseudogene whereas NANOG-ch5 is active in Capra hircus. Bioinformatics analysis indicated that the 5' upstream region of NANOG-ch5 does not have typical CpG islands but contains several CG enrichment regions and several LIN28A binding sites. Deletion analysis suggested that NANOG-ch5 promoter can be activated by LIN28A directly binding to the site -210 but not by the indirect effect from the inhibition of let-7g, which is known to be downregulated by LIN28A. Mechanistically, LIN28A recruits and interacts with 5-methylcytosine-dioxygenase Ten-Eleven translocation 1 (TET1) to NANOG-ch5 gene promoter binding sites to orchestrate 5-methylcytosine and 5-hydroxymethylcytosine dynamics. These results revealed the role of LIN28A in NANOG transcriptional regulation via epigenetic DNA modifications to maintain the stemness of GmGSC.

8.
Aging (Albany NY) ; 10(10): 2954-2972, 2018 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-30362962

RESUMO

Transplantation of adipose-derived mesenchymal stem cells (ADMSCs) can aid in the treatment of numerous diseases in animals. However, natural aging during in vitro expansion of ADMSCs prior to their use in transplantation restricts their beneficial effects. Melatonin is reported to exert biorhythm regulation, anti-oxidation, and anti-senescence effects in various animal and cell models. Herein, by using a senescent canine ADMSCs (cADMSCs) cell model subjected to multiple passages in vitro, we investigated the effects of melatonin on ADMSCs senescence. We found that melatonin alleviates endoplasmic reticulum stress (ERS) and cell senescence. MT1/MT2 melatonin receptor inhibitor, luzindole, diminished the mRNA expression levels and rhythm expression amplitude of Bmal1 and Nrf2 genes. Nrf2 knockdown blocked the stimulatory effects of melatonin on endoplasmic reticulum-associated degradation (ERAD)-related gene expression and its inhibitory effects on ERS-related gene expression. At the same time, the inhibitory effects of melatonin on the NF-κB signaling pathway and senescence-associated secretory phenotype (SASP) were blocked by Nrf2 knockdown in cADMSCs. Melatonin pretreatment improved the survival of cADMSCs and enhanced the beneficial effects of cADMSCs transplantation in canine acute liver injury. These results indicate that melatonin activates Nrf2 through the MT1/MT2 receptor pathway, stimulates ERAD, inhibits NF-κB and ERS, alleviates cADMSCs senescence, and improves the efficacy of transplanted cADMSCs.


Assuntos
Tecido Adiposo/citologia , Senescência Celular/efeitos dos fármacos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Melatonina/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Senescência Celular/genética , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Doença Hepática Induzida por Substâncias e Drogas/cirurgia , Modelos Animais de Doenças , Cães , Estresse do Retículo Endoplasmático/genética , Degradação Associada com o Retículo Endoplasmático/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Fator 2 Relacionado a NF-E2/genética , NF-kappa B/metabolismo , Receptor MT1 de Melatonina/agonistas , Receptor MT1 de Melatonina/metabolismo , Receptor MT2 de Melatonina/agonistas , Receptor MT2 de Melatonina/metabolismo , Transdução de Sinais/efeitos dos fármacos
9.
Cell Death Dis ; 9(10): 968, 2018 09 20.
Artigo em Inglês | MEDLINE | ID: mdl-30237484

RESUMO

Diabetes mellitus affects a large number of men of reproductive age and it usually leads to serious reproductive disorders. However, the underlying mechanisms and specific therapies still remain largely unknown. We observed Leydig cell loss in the testes of diabetic mice. Continuous high glycemic status of testes stimulated expression of Caspase12, Grp78, and Chop, the three ERS response factors; this might induce cell cycle arrest and apoptosis of Leydig cells in response to ERS. In these diabetic mouse models, melatonin alleviated apoptosis of testicular stromal cell induced by ERS, and promoted SSCs self-renewal by recovering Leydig cells secretion of CSF1 after 8 weeks of treatment. To explore the relationship between CSF-1 and ERS in Leydig cells, we treated Leydig tumor cell line with an activator Tuniamycin and an inhibitor 4-Phenylbutyrate of ERS. Our data showed that the CSF-1 expression in mouse Leydig cell lines decreased six-fold while reversely increasing five-fold in the 4-Phenylbutyrate-treated group. Thus, melatonin likely alleviates the loss of Leydig cells in diabetic testes and provides a healthier niche for SSCs to self-renew and continually provide healthy sperm for male fertility.


Assuntos
Células Intersticiais do Testículo/efeitos dos fármacos , Melatonina/uso terapêutico , Espermatogônias/efeitos dos fármacos , Células-Tronco Germinativas Adultas/citologia , Células-Tronco Germinativas Adultas/efeitos dos fármacos , Células-Tronco Germinativas Adultas/metabolismo , Animais , Apoptose/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Células Intersticiais do Testículo/citologia , Células Intersticiais do Testículo/metabolismo , Fator Estimulador de Colônias de Macrófagos/genética , Fator Estimulador de Colônias de Macrófagos/metabolismo , Masculino , Camundongos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Espermatogônias/citologia , Espermatogônias/metabolismo , Testículo/citologia , Testículo/efeitos dos fármacos , Testículo/metabolismo
10.
J Cell Physiol ; 234(1): 915-926, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-30069947

RESUMO

Self-renewal and differentiation of male germline stem cells (mGSCs) provide the basic function for continual spermatogenesis. Studies of in vitro culture of germline stem cells are important and meaningful for basic biological research and practical application. Growth factors, such as GDNF, bFGF, CSF1, and EGF, could maintain the self-renewal of mGSCs. Insulin-like growth factor 1 (IGF-1), an important growth factor, and its pathway have been reported to maintain the survival of several types of stem cells and play important roles in male reproduction. However, the mechanism through which the IGF-1 pathway acts to regulate the self-renewal of mGSCs remains unclear. We analyzed the effect of IGF-1 on the proliferation and apoptosis of bovine mGSCs. We evaluated the expression profile of long noncoding RNA (LncRNA) H19 in bovine and mouse tissues. Moreover, we investigated whether LncRNA H19 could regulate the IGF-1 pathway. Results showed that IGF-1 could activate the phosphorylation of AKT and ERK signaling pathways, and the IGF-1 pathway played an important role in regulating the proliferation and apoptosis of bovine mGSCs. The proliferation rate of mGSCs decreased, whereas the apoptosis rate of mGSCs increased when the IGF-1 receptor (IGF-1R) was blocked using the IGF-1R-specific inhibitor (picropodophyllin). LncRNA H19 could regulate the IGF-1 signaling pathway and, consequently, the proliferation and apoptosis of mGSCs. The number of cells in the seminiferous tubule decreased when H19 was interfered by injecting a virus-containing supernatant. Hence, LncRNA H19 participated in the regulation of the proliferation and apoptosis of mGSCs via the IGF-1 signaling pathway.


Assuntos
Fator de Crescimento Insulin-Like I/genética , RNA Longo não Codificante/genética , Espermatogênese/genética , Células-Tronco/citologia , Animais , Apoptose/genética , Bovinos , Diferenciação Celular/genética , Proliferação de Células/genética , Células Germinativas/citologia , Células Germinativas/metabolismo , Masculino , Camundongos , Transdução de Sinais/genética , Células-Tronco/metabolismo , Testículo/crescimento & desenvolvimento , Testículo/metabolismo
11.
J Environ Manage ; 226: 30-36, 2018 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-30107308

RESUMO

Certain sectors and paths along supply chains play a critical role in climate change mitigation. We develope a consumption-based framework, which combines input-output analysis, a power-of-pull approach and structural path analysis, and applied it to supply chain networks derived from 2010 and 2012 Jing-Jin-Ji interregional input-output tables. The aim of this study is to identify (1) the key economic sectors for controlling carbon emissions and their changes, (2) the critical directions from a carbon-pulling sector to the emissions of key economic sectors, and (3) the paths with the largest carbon emissions flux in these critical directions. Our results show that the key sectors are from Hebei and Tianjin, more concentrated in Hebei. Most sectors have the largest pulling power over their own carbon emissions, and within-region connections dominated in the emission network, with a stronger tie between Beijing and the other two regions. Critical paths along carbon-pulling directions are located in tiers 0 and 1. Our framework can provide new insight into the creation of carbon emissions control policies.


Assuntos
Dióxido de Carbono/economia , Mudança Climática , Fontes Geradoras de Energia/economia , Pequim , Carbono
12.
ACS Appl Mater Interfaces ; 10(18): 15691-15696, 2018 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-29667402

RESUMO

It is of great significance to seek high-performance solid electrolytes via a facile chemistry and simple process for meeting the requirements of solid batteries. Previous reports revealed that ion conducting pathways within ceramic-polymer composite electrolytes mainly occur at ceramic particles and the ceramic-polymer interface. Herein, one facile strategy toward ceramic particles' alignment and assembly induced by an external alternating-current (AC) electric field is presented. It was manifested by an in situ optical microscope that Li1.3Al0.3Ti1.7(PO4)3 particles and poly(ethylene glycol) diacrylate in poly(dimethylsiloxane) (LATP@PEGDA@PDMS) assembled into three-dimensional connected networks on applying an external AC electric field. Scanning electron microscopy revealed that the ceramic LATP particles aligned into a necklacelike assembly. Electrochemical impedance spectroscopy confirmed that the ionic conductivity of this necklacelike alignment was significantly enhanced compared to that of the random one. It was demonstrated that this facile strategy of applying an AC electric field can be a very effective approach for architecting three-dimensional lithium-ion conductive networks within solid composite electrolyte.

13.
J Clin Endocrinol Metab ; 103(6): 2157-2166, 2018 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-29590425

RESUMO

Context: Women with diminished ovarian reserve (DOR) have reduced fertility, cardiovascular events, and osteoporosis. Although differential microRNA (miRNA) expression has been described in several ovarian disorders, little is known about the role of miRNAs in the pathogenesis of DOR. Objective: Identify differentially expressed miRNAs in DOR and explore the role of miR-106a in human granulosa cell proliferation. Design: miRNA microarray (n = 3) and quantitative reverse transcription polymerase chain reaction (n = 30) were used to examine miRNA expression in serum and granulosa cells from normal-cycling and women with DOR. Primary human granulosa cells were treated alone or in combination with miR-106a mimic, miR-106a inhibitor, apoptosis signal-regulating kinase 1 (ASK1) small interfering RNA (siRNA), or p38 mitogen-activated protein kinase (MAPK) inhibitor (SB203580) before assessment of cell viability and apoptosis. Western blot was used to measure ASK1 protein and phosphorylation/activation of p38 MAPK. Binding of miR-106a to ASK1 mRNA was examined by 3' untranslated region (3'UTR) luciferase analysis. Results: Fifteen miRNAs were differentially expressed (n = 30), and miR-106a was downregulated in serum and granulosa cells of women with DOR. miR-106a mimic increased cell viability and attenuated apoptosis, whereas the converse occurred following treatment with miR-106a inhibitor. miR-106a suppressed ASK1 expression by directly targeting its 3'UTR. miR-106a inhibitor increased p38 MAPK phosphorylation/activation, and this effect was abolished by treatment with ASK1 siRNA. Whereas knockdown of ASK1 abolished the effects of miR-106a inhibitor on cell viability/apoptosis, pretreatment with SB203580 did not significantly alter the effects of miR-106a inhibitor. Conclusions: Downregulation of miR-106a may contribute to the pathogenesis of DOR by reducing granulosa cell viability and promoting apoptosis via enhanced ASK1 signaling.


Assuntos
Sobrevivência Celular/efeitos dos fármacos , Regulação para Baixo , Células da Granulosa/metabolismo , Infertilidade Feminina/metabolismo , MicroRNAs/metabolismo , Reserva Ovariana/genética , Apoptose/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Feminino , Regulação da Expressão Gênica , Células da Granulosa/efeitos dos fármacos , Humanos , Imidazóis/farmacologia , Infertilidade Feminina/genética , MAP Quinase Quinase Quinase 5/genética , MAP Quinase Quinase Quinase 5/metabolismo , MicroRNAs/genética , Fosforilação/efeitos dos fármacos , Piridinas/farmacologia , RNA Interferente Pequeno , Transdução de Sinais/efeitos dos fármacos
14.
Cytotechnology ; 70(2): 843-854, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29372468

RESUMO

Germline stem cells (GSCs) play an indispensable role in establishing the fertility of an organism. The isolation and culture of adult female GSCs (FGSCs) have provided a robust foundation to study the development of female germ cells in rodents. However, many problems still need to be identified, such as the origin and location of FGSCs and the specific markers for screening. In this study, we acquired FGSCs that stably expressed Oct4 from Oct4 promoter-GFP transgenic mouse ovarian surface epithelium and cortical layer, and identified the cells possessing the representative features including the expression of GSCs marker genes and the potentiality of differentiation into all three germ layers in vitro. Moreover, rapamycin was confirmed to promote proliferation of mouse FGSCs and inhibit the differentiation capability in vivo. In addition to the reported disinfection function, rapamycin inhibited the activation of primordial follicles, as the inhibitor of mechanistic target of rapamycin pathway. These results will contribute to the study on folliculogenesis or oogenesis mechanism and have important implications on developing new technology and therapeutic approach in medicine for premature ovarian failure, infertility and even ovary remodelling in future.

15.
J Cell Physiol ; 233(3): 2537-2548, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28777437

RESUMO

The protein encoded by double sex and mab-3 related transcription factor 1 (Dmrt1) gene contains a double sex/mab-3 domain, which was considered as one of the most conservative structures in sex determination. However, its effect on spermatogenesis of dairy goat spermatogonial stem cells (SSCs) remains to be clarified. For the first time, the roles of Dmrt1 in spermatogenesis of livestock are highlighted. Here, we investigated the expression pattern of Dmrt1 in the testes of dairy goats. Dmrt1 primarily located in undifferentiated SSCs. Moreover, Dmrt1 enhanced differentiation and proliferation of mGSCs. On the contrary, the level of meiosis was down-regulated, as Dmrt1 determines whether SSCs undergo mitosis and spermatogonial differentiation or meiosis. In the busulfan-treated mice testes, Dmrt1 repair germ cell damage was emphasized as well. Our results exposed that Dmrt1 maintenance mGSCs in two ways: facilitating proliferation and self-renewal of SSCs; and reducing the inflammatory response caused by reproductive injury. These findings identify a central role for Dmrt1 in controlling population stability and injury restoring of SSCs.


Assuntos
Células-Tronco Germinativas Adultas/metabolismo , Diferenciação Celular , Proliferação de Células , Autorrenovação Celular , Indústria de Laticínios , Cabras/metabolismo , Espermatogênese , Fatores de Transcrição/metabolismo , Células-Tronco Germinativas Adultas/efeitos dos fármacos , Células-Tronco Germinativas Adultas/patologia , Animais , Bussulfano/toxicidade , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Autorrenovação Celular/efeitos dos fármacos , Células Cultivadas , Regulação da Expressão Gênica no Desenvolvimento , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Cabras/genética , Fator Estimulador de Colônias de Macrófagos/metabolismo , Masculino , Meiose , Mitose , Proteína com Dedos de Zinco da Leucemia Promielocítica/metabolismo , Transdução de Sinais , Espermatogênese/efeitos dos fármacos , Fatores de Transcrição/genética , Transfecção , Tretinoína/farmacologia
16.
Antioxid Redox Signal ; 28(5): 385-400, 2018 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-28027652

RESUMO

AIMS: Many men endure immunosuppressive or anticancer treatments that contain alkylating agents before the age of sexual maturity, especially the increasing number of preadolescent males who undergo busulfan treatment for myeloablative conditioning before hematopoietic stem cell transplantation. Before sperm production, there are no sperm available for cryopreservation. Thus, it is necessary to identify a solution to ameliorate the busulfan-induced damage of spermatogonial stem cells (SSCs). RESULTS: In this study, we demonstrated that melatonin relieved the previously described SSC loss and apoptosis in mouse testes. Melatonin increased the expression of manganese superoxide dismutase (MnSOD), which regulated the production of busulfan-induced reactive oxygen species (ROS). Moreover, melatonin promoted sirtuin type 1 (SIRT1) expression. SIRT1 participated in the deacetylation of p53, which promotes p53 ubiquitin degradation. Decreased concentrations of deacetylated p53 resulted in spermatogonial cell resistance to apoptosis. Acute T cell leukemia cell assay demonstrated that melatonin does not affect busulfan-induced cancer cell apoptosis and ROS. INNOVATION: The current evidence suggests that melatonin may alleviate the side effects of alkylating drugs, such as busulfan. CONCLUSION: Melatonin promoted MnSOD and SIRT1 expression, which successfully ameliorated busulfan-induced SSC apoptosis caused by high concentrations of ROS and p53. Antioxid. Redox Signal. 28, 385-400.


Assuntos
Melatonina/administração & dosagem , Sirtuína 1/genética , Superóxido Dismutase/genética , Testículo/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Bussulfano/efeitos adversos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Humanos , Masculino , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Espermatogônias/efeitos dos fármacos , Espermatogônias/crescimento & desenvolvimento , Espermatogônias/patologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/patologia , Testículo/crescimento & desenvolvimento , Testículo/patologia , Proteína Supressora de Tumor p53/genética , Ubiquitina/genética
17.
Cell Physiol Biochem ; 44(6): 2407-2421, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29268276

RESUMO

BACKGROUND/AIMS: Busulfan is commonly used for cancer chemotherapy. Although it has the advantage of increasing the survival rate of patients, it can cause male infertility via damaging the testes and reducing sperm counts. Therefore, the underlying mechanism should be explored, and new agents should be developed to protect the male reproductive system from busulfan-induced damage. Endoplasmic reticulum stress (ERS) is considered a key contributor to numerous pathologies. Despite several studies linking ERS to toxicants, studies have yet to determine whether ERS is a contributing factor to busulfan-induced testicular damage. Melatonin is a well-known broad-spectrum antioxidant, anti-inflammatory and antitumour agent, but the effects of melatonin on busulfan-induced ERS in mouse testes damage are less documented. METHODS: The effects of melatonin were measured by immunofluorescence staining, Western blot, qRT-PCR analysis and flow cytometry assay. The underlying mechanism was investigated by measuring ERS. RESULTS: We found that ERS was strongly activated in mouse testes (in vivo) and the C18-4 cell line (in vitro) after busulfan administration. ERS-related apoptosis proteins such as caspase-12, CHOP and caspase-3 were activated, and the expression of apoptotic proteins such as P53 and PUMA were upregulated. Furthermore, we investigated whether melatonin reduced the extent of damage to mouse testes and improved the survival rates of busulfan-treated mice. When exploring the underlying mechanisms, we found melatonin could counteract ERS by decreasing the expression levels of the ERS markers GRP78, ATF6, pIRE1 and XBP1 in mouse testes and mouse SSCs (C18-4 cells). Moreover, it blocked the activation of ERS-related apoptosis proteins caspase-12, CHOP and caspase-3 and suppressed P53 and PUMA expression stimulated by busulfan both in vivo and in vitro. CONCLUSION: Our results demonstrate that ERS is an important mediator for busulfan-induced apoptosis. The attenuation of ERS by melatonin can prevent busulfan-treated SSCs apoptosis and protect busulfan-treated testes from damage. Thus, this study suggests that melatonin may alleviate the side effects of busulfan for male patients during clinical treatment.


Assuntos
Antineoplásicos Alquilantes/efeitos adversos , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Bussulfano/efeitos adversos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Melatonina/farmacologia , Testículo/efeitos dos fármacos , Animais , Linhagem Celular , Masculino , Camundongos , Camundongos Endogâmicos ICR , Testículo/citologia , Testículo/patologia
18.
PLoS One ; 12(10): e0187159, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29073244

RESUMO

Porcine pancreatic stem cells (PSCs) are considered promising transplant materials that may be used to treat diabetes, but some problems, such as insufficient cell number and low differentiation efficiency, should be solved before its clinical application. Resveratrol is a natural polyphenolic compound that can alleviate the complications of diabetes. In this study, we aimed to explore the specific effect of resveratrol on porcine PSCs. We treated porcine PSCs with 10 µM, 25 µM resveratrol to explore the effect of resveratrol on porcine PSCs. We found that 10 µM resveratrol improved the proliferation of porcine PSCs, increased the expression of A-ß-catenin (active ß-catenin), Pcna, C-Myc, Bcl-2 and sirtuin-1 (Sirt1), and decreased the expression of P53, Caspase3. While 25 µM resveratrol had almost opposite effect compared with 10 µM resveratrol group. The utilization of Dickkopf-related protein 1 (DKK1, Wnt signaling pathway inhibitor) and nicotinamide (Sirt1 inhibitor) suggested that resveratrol regulated cell proliferation by controlling Wnt signaling pathway and this effect was mediated by Sirt1. Our results further revealed that 10 µM resveratrol promoted the formation of ß-like cells regulated by Wnt/ß-catenin signal pathway. Relatively low-dose resveratrol could improve porcine PSCs fate. It lays theoretical foundation for diabetes treatment with cell transplantation in future.


Assuntos
Pâncreas/efeitos dos fármacos , Sirtuína 1/metabolismo , Células-Tronco/efeitos dos fármacos , Estilbenos/farmacologia , Via de Sinalização Wnt/efeitos dos fármacos , Animais , Linhagem da Célula , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Glucose/administração & dosagem , Pâncreas/citologia , Pâncreas/metabolismo , Resveratrol , Células-Tronco/metabolismo , Suínos
19.
Sci Rep ; 7(1): 12174, 2017 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-28939897

RESUMO

Bitter taste receptors (TAS2Rs) have attracted a great deal of interest because of their recently described bronchodilator and anti-inflammatory properties. The aim of this study was to identify natural direct TAS2R14 agonists from Radix Bupleuri that can inhibit mast cell degranulation. A ligand-based virtual screening was conducted on a library of chemicals contained in compositions of Radix Bupleuri, and these analyses were followed by cell-based functional validation through a HEK293-TAS2R14-G16gust44 cell line and IgE-induced mast cell degranulation assays, respectively. Saikosaponin b (SSb) was confirmed for the first time to be a specific agonist of TAS2R14 and had an EC50 value of 4.9 µM. A molecular docking study showed that SSb could directly bind to a TAS2R14 model through H-bond interactions with Arg160, Ser170 and Glu259. Moreover, SSb showed the ability to inhibit IgE-induced mast cell degranulation, as measured with a ß-hexosaminidase release model and real-time cell analysis (RTCA). In a cytotoxicity bioassay, SSb showed no significant cytotoxicity to HEK293 cells within 24 hours. This study demonstrated that SSb is a direct TAS2R14 agonist that inhibit IgE-induced mast cell degranulation. Although the target and in vitro bioactivity of SSb were revealed in this study, it still need in vivo study to further verify the anti-asthma activity of SSb.


Assuntos
Bupleurum/química , Ácido Oleanólico/análogos & derivados , Receptores Acoplados a Proteínas-G/agonistas , Receptores Acoplados a Proteínas-G/metabolismo , Saponinas/farmacologia , Sítios de Ligação , Degranulação Celular/efeitos dos fármacos , Linhagem Celular , Células HEK293 , Humanos , Ligações de Hidrogênio , Imunoglobulina E/metabolismo , Mastócitos/efeitos dos fármacos , Simulação de Acoplamento Molecular , Ácido Oleanólico/química , Ácido Oleanólico/farmacologia , Ligação Proteica , Receptores Acoplados a Proteínas-G/química , Saponinas/química
20.
R Soc Open Sci ; 4(8): 170512, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28878998

RESUMO

Flame-retardant polyvinyl alcohol (PVA) membranes with high transparency and flexibility were prepared by mixing an aqueous solution of a phosphorus-containing acrylic acid (AOPA) with PVA. The reaction between AOPA and PVA, the transparency, the crystallinity and the flexibility of the membrane were investigated with Fourier transform infrared spectrometry (FTIR), UV-vis light transmittance, X-ray diffraction and tensile tests, respectively. The limited oxygen index (LOI) and vertical flame (UL 94 VTM), microscale combustion calorimetry, thermogravimetric analysis (TGA) and TGA-FTIR were employed to evaluate the flame retardancy as well as to reveal the corresponding mechanisms. Results showed that PVA containing 30 wt% of AOPA can reach the UL 94 VTM V0 rating with an LOI of 27.3% and retain 95% of the original transparency of pure PVA. Adding AOPA reduces crystallinity of PVA, while the flexibility is increased. AOPA depresses the thermal degradation of PVA and promotes char formation during combustion. The proposed decomposition mechanism indicates that AOPA acts mainly in the condensed phase.

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