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1.
Cereb Cortex ; 33(8): 4293-4304, 2023 04 04.
Artigo em Inglês | MEDLINE | ID: mdl-36030380

RESUMO

Neocortical vasoactive intestinal polypeptide-expressing (VIP+) interneurons display highly diverse morpho-electrophysiological and molecular properties. To begin to understand the function of VIP+ interneurons in cortical circuits, they must be clearly and comprehensively classified into distinct subpopulations based on specific molecular markers. Here, we utilized patch-clamp RT-PCR (Patch-PCR) to simultaneously obtain the morpho-electric properties and mRNA profiles of 155 VIP+ interneurons in layers 2 and 3 (L2/3) of the mouse somatosensory cortex. Using an unsupervised clustering method, we identified 3 electrophysiological types (E-types) and 2 morphological types (M-types) of VIP+ interneurons. Joint clustering based on the combined electrophysiological and morphological features resulted in 3 morpho-electric types (ME-types). More importantly, we found these 3 ME-types expressed distinct marker genes: ~94% of Sncg+ cells were ME-type 1, 100% of Mybpc1+ cells were ME-type 2, and ~78% of Parm1+ were ME-type 3. By clarifying the properties of subpopulations of cortical L2/3 VIP+ interneurons, this study establishes a basis for future investigations aiming to elucidate their physiological roles.


Assuntos
Córtex Somatossensorial , Peptídeo Intestinal Vasoativo , Animais , Camundongos , Fenômenos Eletrofisiológicos , Interneurônios/fisiologia , Córtex Somatossensorial/fisiologia , Peptídeo Intestinal Vasoativo/metabolismo , Proteínas de Neoplasias/metabolismo , gama-Sinucleína/metabolismo , Proteína de Ligação a Androgênios/metabolismo
2.
Int Immunopharmacol ; 88: 106940, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32916626

RESUMO

AIM: To clarify the regulatory mechanisms of lacrimal androgen-binding proteins (ABPs) in mice with keratitis caused by Aspergillus fumigatus (A. fumigatus). METHODS: Mouse models of A. fumigatus keratitis were established. Lacrimal glands were removed after 24 h for general and histological comparison. Lacrimal ABPs were detected by qRT-PCR and quantitative proteomic analysis, or were detected by qRT-PCR after subconjunctival or lacrimal gland injection with dexamethasone. Unique inflammatory factors were detected by qRT-PCR, Western blot and/or immunofluorescence. Interleukin-1ß (IL-1ß) was injected into the lacrimal gland to explore the relationship between IL-1ß and lacrimal ABPs. RESULTS: The lacrimal glands of mice with fungal keratitis were larger than normal mice and these structures became disorganized. Moreover, the expression of ABP ε and ABP δ were increased. Subconjunctival injection with dexamethasone could reduce the size of the lacrimal gland and increase the expression of ABP ε and ABP δ, while lacrimal gland injection with dexamethasone had no obvious effects. The expression of IL-1ß in the lacrimal gland of mice with A. fumigatus keratitis was increased. When IL-1ß was injected into the lacrimal gland, the lacrimal gland enlarged and the expression of ABP ε and ABP δ decreased. CONCLUSION: Lacrimal glands contributed to protection in fungal keratitis, which was not due to the involvement of inflammatory cells in mice. ABP δ and ABP ε of mice were involved in reducing the severity of corneal damage in mice with A. fumigatus keratitis. Moreover, the expression of IL-1ß and ABP δ and ABP ε were intrinsically linked.


Assuntos
Proteína de Ligação a Androgênios/metabolismo , Aspergilose/metabolismo , Aspergillus fumigatus , Ceratite/metabolismo , Aparelho Lacrimal/metabolismo , Proteína de Ligação a Androgênios/genética , Animais , Aspergilose/genética , Citocinas/genética , Citocinas/metabolismo , Feminino , Ceratite/genética , Masculino , Camundongos Endogâmicos C57BL
3.
J Cell Biochem ; 120(10): 18041-18052, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31297877

RESUMO

Owing to the high morbidity and mortality, novel biomarkers in the occurrence and development of colorectal cancer (CRC) are needed nowadays. In this study, the CRC-related datasets were downloaded from the Gene Expression Omnibus (GEO) database and The Cancer Genome Atlas (TCGA) database. After screening the differentially expressed genes (DEGs) in R software, a total of 238 upregulated and 199 downregulated DEGs were revealed simultaneously. Then the Kaplan-Meier survival analysis and Cox regression analysis were used to reveal the prognostic function of these DEGs. Neurexophilin and PC-esterase domain family member 4 (NXPE4) and prostate androgen-regulated mucin-like protein 1 (PARM1) were two outstanding independent overall survival (OS) and relapse-free survival (RFS) prognostic genes of CRC in TCGA database. We next verified the expression of NXPE4 and PARM1 messenger RNA (mRNA) levels were significantly lower in CRC tumor tissue than in the adjacent noncancerous tissue in our clinical samples, and NXPE4 mRNA expression level was related to the tumor location and tumor size, while PARM1 was related to tumor location, lymph nodes metastasis, and tumor size. This study demonstrated that NXPE4 and PARM1 might be two potential novel prognostic biomarkers for CRC.


Assuntos
Biomarcadores Tumorais/genética , Neoplasias Colorretais/genética , Glicoproteínas/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Neuropeptídeos/genética , Idoso , Proteína de Ligação a Androgênios/metabolismo , Biomarcadores Tumorais/metabolismo , Intervalo Livre de Doença , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Ontologia Genética , Glicoproteínas/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Neuropeptídeos/metabolismo , Prognóstico , Modelos de Riscos Proporcionais , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
4.
Environ Toxicol Pharmacol ; 63: 84-91, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30189373

RESUMO

Arsenic (As) has been recognized as a cause of male reproductive toxicity. However, effects of long-term arsenic exposure (puberty-adult) on spermatogenesis, testosterone synthesis, and the expression of androgen binding protein (ABP) and Ddx3y remain unclear. The objective of this investigation was to explore these effects and the underlying mechanisms. Male mice were treated with 5 and 50 ppm arsenic for 6 months via drinking water. The results showed that arsenic reduced sperm count and sperm motility and enhanced the abnormal sperm percentage. The decrease in the number of spermatogenic cells and sperm in seminiferous tubules and the decline in the Johnsen score were observed in both arsenic-treated groups, suggesting spermatogenesis disorders. Moreover, arsenic diminished serum testosterone, along with the reduced expression of luteinizing hormone receptor (LHR), steroidogenic acute regulatory protein (StAR) and 17-ß-hydroxysteroid dehydrogenase (17ß-HSD) genes. Arsenic also down-regulated mRNA levels of ABP and Ddx3y in a dose-dependent manner. Meanwhile, the protein levels of StAR, 17ß-HSD and Ddx3y were significantly reduced in arsenic-treated groups. Taken together, these results suggest that the reduced testosterone through inhibition of the expression of multiple genes responsible for the biosynthesis, the damaged androgen homeostasis partially via lessening the expression levels of the ABP gene and the down-regulated expression of Ddx3y, may contribute to spermatogenesis disorders in mice exposed to arsenic.


Assuntos
Proteína de Ligação a Androgênios/genética , Arsênio/toxicidade , RNA Helicases DEAD-box/genética , Regulação para Baixo , Antígenos de Histocompatibilidade Menor/genética , Espermatogênese/efeitos dos fármacos , Testosterona/sangue , 17-Hidroxiesteroide Desidrogenases/genética , 17-Hidroxiesteroide Desidrogenases/metabolismo , Proteína de Ligação a Androgênios/metabolismo , Animais , RNA Helicases DEAD-box/metabolismo , Relação Dose-Resposta a Droga , Água Potável , Masculino , Camundongos , Antígenos de Histocompatibilidade Menor/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Receptores do LH/genética , Receptores do LH/metabolismo , Contagem de Espermatozoides , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos
5.
Mol Med Rep ; 16(5): 6443-6458, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28901490

RESUMO

Prostate cancer (PCa) poses a high risk to older men and it is the second most common type of male malignant tumor in western developed countries. Additionally, there is a lack of effective therapies for PCa at advanced stages. Novel treatment strategies such as adenovirus­mediated gene therapy and virotherapy involve the expression of a specific therapeutic gene to induce death in cancer cells, however, wild­type adenoviruses are also able to infect normal human cells, which leads to undesirable toxicity. Various PCa­targeting strategies in adenovirus­mediated therapy have been developed to improve tumor­targeting effects and human safety. The present review summarizes the relevant knowledge regarding available adenoviruses and PCa­targeting strategies. In addition, future directions in this area are also discussed. In conclusion, although they remain in the early stages of basic research, adenovirus­mediated gene therapy and virotherapy are expected to become important therapies for tumors in the future due to their potential targeting strategies.


Assuntos
Adenoviridae/genética , Genes Virais , Terapia Genética/métodos , Terapia de Alvo Molecular/métodos , Terapia Viral Oncolítica/métodos , Neoplasias da Próstata/terapia , Adenoviridae/metabolismo , Proteína de Ligação a Androgênios/genética , Proteína de Ligação a Androgênios/metabolismo , Antígenos de Superfície/genética , Antígenos de Superfície/metabolismo , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismo , Deleção de Genes , Glutamato Carboxipeptidase II/genética , Glutamato Carboxipeptidase II/metabolismo , Humanos , Masculino , Regiões Promotoras Genéticas , Próstata/metabolismo , Próstata/patologia , Próstata/virologia , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Neoplasias da Próstata/virologia
6.
PLoS One ; 12(5): e0178041, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28542629

RESUMO

The goal was to gain understanding of how 12 genes containing SNP previously related to embryo competence to become a blastocyst (BRINP3, C1QB, HSPA1L, IRF9, MON1B, PARM1, PCCB, PMM2, SLC18A2, TBC1D24, TTLL3 and WBP1) participate in embryonic development. Gene expression was evaluated in matured oocytes and embryos. BRINP3 and C1QB were not detected at any stage. For most other genes, transcript abundance declined as the embryo developed to the blastocyst stage. Exceptions were for PARM1 and WBP1, where steady-state mRNA increased at the 9-16 cell stage. The SNP in WBP1 caused large differences in the predicted three-dimensional structure of the protein while the SNP in PARM1 caused smaller changes. The mutation in WBP1 causes an amino acid substitution located close to a P-P-X-Y motif involved in protein-protein interactions. Moreover, the observation that the reference allele varies between mammalian species indicates that the locus has not been conserved during mammalian evolution. Knockdown of mRNA for WBP1 decreased the percent of putative zygotes becoming blastocysts and reduced the number of trophectoderm cells and immunoreactive CDX2 in the resulting blastocysts. WBP1 is an important gene for embryonic development in the cow. Further research to identify how the SNP in WBP1 affects processes leading to differentiation of the embryo into TE and ICM lineages is warranted.


Assuntos
Blastocisto/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Sequência de Aminoácidos , Proteína de Ligação a Androgênios/química , Proteína de Ligação a Androgênios/genética , Proteína de Ligação a Androgênios/metabolismo , Animais , Blastocisto/citologia , Bovinos , Células Cultivadas , Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário , Fertilização in vitro , Regulação da Expressão Gênica no Desenvolvimento , Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Peptídeos e Proteínas de Sinalização Intracelular/classificação , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Oligonucleotídeos Antissenso/metabolismo , Oócitos/citologia , Oócitos/metabolismo , Filogenia , Polimorfismo de Nucleotídeo Único , Estrutura Terciária de Proteína , RNA Mensageiro/metabolismo , Alinhamento de Sequência
7.
Urol J ; 14(2): 3023-3027, 2017 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-28299766

RESUMO

PURPOSE: Varicocele is the most common risk factor for male infertility, however, not all males with varicocele experience infertility. In fact, most patients with varicocele have normal spermatogenesis. The molecular mechanism of varicocele-associated infertility is yet to be completely understood. The aim of this study is to assess the association of a number of fertility regulatory factors on varicocele associated infertility and to throw light on the mechanism of varicocele-associated infertility. MATERIALS AND METHODS: Semen from 30 infertile patients with varicocele and 30 fertile men with varicocele were collected. The concentrations of the following factors in seminal plasma were determined by ELISA: follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), androgen binding protein (ABP), transferrin (Trf), inhibin B (INHB) and stem cell factor (SCF). The expression level of c-kit in seminal precipitate of patients with varicocele was detected by real-time PCR. RESULTS: The concentrations of sexual hormones, FSH, LH and T, had no differences between infertile patients with varicocele and fertile men with varicocele (P > 0.05). Factors secreted by Sertoli cells, ABP, Trf, INHB andSCF, showed no significant differences between the two groups (P > 0.05). Interestingly, the expression of c-kit was significant higher in infertile patients with varicocele than that in fertile men with varicocele (P < 0.01). CONCLUSION: Neither the sexual hormones nor the Sertoli cells was responsible for the infertility induced by varicocele.The aberrant expression of c-kit in infertile patients with varicocele may provide new insight into the mechanism of varicocele-associated infertility.


Assuntos
Infertilidade Masculina/genética , Infertilidade Masculina/metabolismo , Proteínas Proto-Oncogênicas c-kit/genética , Sêmen/metabolismo , Varicocele/genética , Varicocele/metabolismo , Adulto , Proteína de Ligação a Androgênios/metabolismo , Estudos de Casos e Controles , Hormônio Foliculoestimulante/metabolismo , Expressão Gênica , Humanos , Infertilidade Masculina/etiologia , Inibinas/metabolismo , Hormônio Luteinizante/metabolismo , Masculino , Análise do Sêmen , Fator de Células-Tronco/metabolismo , Testosterona/metabolismo , Transferrina/metabolismo , Varicocele/complicações , Adulto Jovem
8.
Genetics ; 205(4): 1517-1527, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-28159752

RESUMO

The house mouse Androgen-binding protein (Abp) gene family is comprised of 64 paralogs, 30 Abpa and 34 Abpbg, encoding the alpha (ABPA) and beta-gamma (ABPBG) protein subunits that are disulfide-bridged to form dimers in secretions. Only 14 Abp genes are expressed in distinct patterns in the lacrimal (11) and submandibular glands (3). We created a knockout mouse line lacking two of the three genes expressed in submandibular glands, Abpa27 and Abpbg27, by replacing them with the neomycin resistance gene. The knockout genotype (-/-) showed no Abpa27 or Abpbg27 transcripts in submandibular gland complementary DNA (cDNA) libraries and there was a concomitant lack of protein expression of ABPA27 and ABPBG27 in the -/- genotype saliva, shown by elimination of these two proteins from the saliva proteome and the loss of cross-reactive material in the acinar cells of the submandibular glands. We also observed a decrease in BG26 protein in the -/- animals, suggesting monomer instability. Overall, we observed no major phenotypic changes in the -/- genotype, compared with their +/+ and +/- siblings raised in a laboratory setting, including normal growth curves, tissue histology, fecundity, and longevity. The only difference is that male and female C57BL/6 mice preferred saliva of the opposite sex containing ABP statistically significantly more than saliva of the opposite sex without ABP in a Y-maze test. These results show for the first time that mice can sense the presence of ABP between saliva targets with and without ABPs, and that they spend more time investigating the target containing ABP.


Assuntos
Proteína de Ligação a Androgênios/genética , Fenótipo , Glândulas Salivares/metabolismo , Proteína de Ligação a Androgênios/metabolismo , Animais , Feminino , Fertilidade , Longevidade , Masculino , Preferência de Acasalamento Animal , Aprendizagem em Labirinto , Camundongos , Proteoma , Saliva/metabolismo
9.
Anim Reprod Sci ; 179: 35-43, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28233584

RESUMO

Podoplanin (PDPN) is a transmembrane receptor which is involved in various physiological and pathological processes, such as cell motility, invasion, tumor metastasis and blood vessels formation. Although there are reports on the involvement of PDPN in Sertoli cells in human and mice, the role of PDPN on the development of bovine Sertoli cells has not been reported. In the present study, Sertoli cells were isolated from 1-day-old bovine testes by two steps enzyme digestion method. Feulgen staining of satellite karyosomes and inhibin immunofluorescence staining suggested that the isolated immature Sertoli cells were very pure. Transfection with overexpression plasmid pBI-CMV3-PDPN and interference shRNA plasmid indicated that PDPN could significantly promote Sertoli cells cycle progression, cells proliferation and androgen-binding protein (ABP) production. Our results indicated that PDPN gene plays a significant role in the proliferation and maturation of bovine Sertoli cells.


Assuntos
Bovinos/fisiologia , Proliferação de Células/fisiologia , Regulação da Expressão Gênica/fisiologia , Glicoproteínas de Membrana/metabolismo , Células de Sertoli/fisiologia , Proteína de Ligação a Androgênios/genética , Proteína de Ligação a Androgênios/metabolismo , Animais , Células Cultivadas , Masculino , Glicoproteínas de Membrana/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Regulação para Cima/fisiologia
10.
Andrologia ; 49(6)2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27659778

RESUMO

The aim was to investigate the effects of long-term heat stress and dietary restriction on the expression of certain genes involving in steroidogenic pathway and small heat-shock proteins (sHSPs) in rat testis. Sprague Dawley rats (n = 24) were equally divided into four groups. Group I and II were kept at an ambient temperature of 22°C, while Groups III and IV were reared at 38°C for 9 weeks. Feed was freely available for Group I and Group III, while Group II and Group IV were fed 60% of the diet consumed by their ad libitum counterparts. At the end of 9 weeks, testicles were collected under euthanasia. Total RNA was isolated from testis tissue samples. Expression profiles of the genes encoding androgen-binding protein, follicle-stimulating hormone receptor, androgen receptor, luteinising hormone receptor, steroidogenic acute regulatory protein (StAR), cyclooxygenase-2 and sHSP genes were assessed at mRNA levels using qPCR. Long-term heat stress decreased the expression of StAR and HspB10 genes while dietary restriction upregulated StAR gene expression. The results suggested that long-term heat stress negatively affected the expression of StAR and HspB10 genes and the dietary restriction was able to reverse negative effect of heat stress on the expression of StAR gene in rat testis.


Assuntos
Restrição Calórica , Regulação da Expressão Gênica , Transtornos de Estresse por Calor/metabolismo , Proteínas de Choque Térmico Pequenas/genética , Testículo/metabolismo , Proteína de Ligação a Androgênios/genética , Proteína de Ligação a Androgênios/metabolismo , Animais , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Proteínas de Choque Térmico Pequenas/metabolismo , Masculino , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Androgênicos/genética , Receptores Androgênicos/metabolismo , Receptores do FSH/genética , Receptores do FSH/metabolismo , Receptores do LH/genética , Receptores do LH/metabolismo
11.
Environ Toxicol Pharmacol ; 42: 146-55, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26851377

RESUMO

The aim of this study was to investigate the effects of ZEA on the cytoskeletal structure, and factors specifically expressed by Sertoli cells. Primary Sertoli cells from rats aged 18-21 days were exposed to increasing ZEA concentrations (0, 5, 10, 20 µg mL(-1)) for 24 h. The results of immunofluorescence showed disruption of α-tubulin filaments and F-actin bundles, and damage to the nucleus of Sertoli cells on exposure to ZEA. In the control group, the protein level expression of androgen-binding protein (ABP), transferrin, vimentin, N-cadherin, and follicle-stimulating hormone receptor (FSHR) were decreased significantly (p<0.05, p<0.01). The mRNA levels of ABP, transferrin, vimentin, N-cadherin, and FSHR varied significantly in the experimental group (p<0.05). The results of enzyme-linked immunosorbent assay indicated a significant decrease in the levels of inhibin-ß and transferrin in the cultural supernatants (p<0.05). Additionally, the ultrastructural analysis indicated the absence of mitochondria and Golgi apparatus, and presence of vacuoles in the cytoplasm. These findings showed that ZEA treatment can damage the cytoskeletal structure and affect specific secretory functions of Sertoli cells, which may be an underlying cause of ZEA-induced reproductive toxicity.


Assuntos
Estrogênios não Esteroides/toxicidade , Zearalenona/toxicidade , Proteína de Ligação a Androgênios/metabolismo , Animais , Caderinas/metabolismo , Inibinas/metabolismo , Masculino , RNA Mensageiro/metabolismo , Ratos , Receptores do FSH/metabolismo , Células de Sertoli/efeitos dos fármacos , Vimentina/metabolismo
12.
Planta Med ; 81(10): 804-12, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26039262

RESUMO

Understanding the molecular mechanism of action of traditional medicines is an important step towards developing marketable drugs from them. Piperine, an active constituent present in the Piper species, is used extensively in Ayurvedic medicines (practiced on the Indian subcontinent). Among others, piperine is known to possess a male contraceptive effect; however, the molecular mechanism of action for this effect is not very clear. In this regard, detailed docking and molecular dynamics simulation studies of piperine with the androgen-binding protein and androgen receptors were carried out. Androgen receptors control male sexual behavior and fertility, while the androgen-binding protein binds testosterone and maintains its concentration at optimal levels to stimulate spermatogenesis in the testis. It was found that piperine docks to the androgen-binding protein, similar to dihydrotestosterone, and to androgen receptors, similar to cyproterone acetate (antagonist). Also, the piperine-androgen-binding protein and piperine-androgen receptors interactions were found to be stable throughout 30 ns of molecular dynamics simulation. Further, two independent simulations for 10 ns each also confirmed the stability of these interactions. Detailed analysis of the piperine-androgen-binding protein interactions shows that piperine interacts with Ser42 of the androgen-binding protein and could block the binding with its natural ligands dihydrotestosterone/testosterone. Moreover, piperine interacts with Thr577 of the androgen receptors in a manner similar to the antagonist cyproterone acetate. Based on the in silico results, piperine was tested in the MDA-kb2 cell line using the luciferase reporter gene assay and was found to antagonize the effect of dihydrotestosterone at nanomolar concentrations. Further detailed biochemical experiments could help to develop piperine as an effective male contraceptive agent in the future.


Assuntos
Alcaloides/química , Alcaloides/farmacologia , Proteína de Ligação a Androgênios/metabolismo , Benzodioxóis/química , Benzodioxóis/farmacologia , Anticoncepcionais Masculinos/farmacologia , Piperidinas/química , Piperidinas/farmacologia , Alcamidas Poli-Insaturadas/química , Alcamidas Poli-Insaturadas/farmacologia , Receptores Androgênicos/metabolismo , Alcaloides/metabolismo , Proteína de Ligação a Androgênios/química , Benzodioxóis/metabolismo , Domínio Catalítico , Linhagem Celular/efeitos dos fármacos , Simulação por Computador , Anticoncepcionais Masculinos/química , Di-Hidrotestosterona/farmacologia , Humanos , Ligação de Hidrogênio , Masculino , Metribolona/química , Metribolona/metabolismo , Metribolona/farmacologia , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Piperidinas/metabolismo , Alcamidas Poli-Insaturadas/metabolismo , Conformação Proteica , Receptores Androgênicos/química , Serina/metabolismo
13.
Sci Rep ; 5: 8894, 2015 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-25745956

RESUMO

Dysregulation of androgen-binding protein (ABP) is associated with a number of endocrine and andrology diseases. However, the ABP metabolism in Sertoli cells is largely unknown. We report that autophagy degrades ABP in rat Sertoli cells, and the autophagic clearance of ABP is regulated by testosterone, which prolongs the ABP biological half-life by inhibiting autophagy. Further studies identified that the autophagic clearance of ABP might be selectively regulated by testosterone, independent of stress (hypoxia)-induced autophagic degradation. These data demonstrate that testosterone up-regulates ABP expression at least partially by suppressing the autophagic degradation. We report a novel finding with respect to the mechanisms by which ABP is cleared, and by which the process is regulated in Sertoli cells.


Assuntos
Proteína de Ligação a Androgênios/metabolismo , Autofagia/fisiologia , Células de Sertoli/citologia , Células de Sertoli/fisiologia , Testosterona/metabolismo , Animais , Células Cultivadas , Masculino , Taxa de Depuração Metabólica , Ratos
14.
PLoS One ; 9(12): e115454, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25531410

RESUMO

The Androgen-binding protein (Abp) region of the mouse genome contains 30 Abpa genes encoding alpha subunits and 34 Abpbg genes encoding betagamma subunits, their products forming dimers composed of an alpha and a betagamma subunit. We endeavored to determine how many Abp genes are expressed as proteins in tears and saliva, and as transcripts in the exocrine glands producing them. Using standard PCR, we amplified Abp transcripts from cDNA libraries of C57BL/6 mice and found fifteen Abp gene transcripts in the lacrimal gland and five in the submandibular gland. Proteomic analyses identified proteins corresponding to eleven of the lacrimal gland transcripts, all of them different from the three salivary ABPs reported previously. Our qPCR results showed that five of the six transcripts that lacked corresponding proteins are expressed at very low levels compared to those transcripts with proteins. We found 1) no overlap in the repertoires of expressed Abp paralogs in lacrimal gland/tears and salivary glands/saliva; 2) substantial sex-limited expression of lacrimal gland/tear expressed-paralogs in males but no sex-limited expression in females; and 3) that the lacrimal gland/tear expressed-paralogs are found exclusively in ancestral clades 1, 2 and 3 of the five clades described previously while the salivary glands/saliva expressed-paralogs are found only in clade 5. The number of instances of extremely low levels of transcription without corresponding protein production in paralogs specific to tears and saliva suggested the role of subfunctionalization, a derived condition wherein genes that may have been expressed highly in both glands ancestrally were down-regulated subsequent to duplication. Thus, evidence for subfunctionalization can be seen in our data and we argue that the partitioning of paralog expression between lacrimal and salivary glands that we report here occurred as the result of adaptive evolution.


Assuntos
Proteína de Ligação a Androgênios/genética , Proteína de Ligação a Androgênios/metabolismo , Genoma , Aparelho Lacrimal/metabolismo , Saliva/metabolismo , Glândula Submandibular/metabolismo , Lágrimas/metabolismo , Proteína de Ligação a Androgênios/classificação , Animais , Western Blotting , Células Cultivadas , Evolução Molecular , Feminino , Perfilação da Expressão Gênica , Aparelho Lacrimal/citologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteômica , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saliva/citologia , Seleção Genética , Glândula Submandibular/citologia
15.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 39(6): 598-604, 2014 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-25011964

RESUMO

OBJECTIVE: To determine the effect of soy isoflavones on cell proliferation and the transcription levels of follicle-stimulating hormone receptor (FSHR), inhibin α (INHα), INHßB, androgen binding protein (ABP), transferrin (Tf) and vimentin in testis sertoli cells in SD rats. METHODS: Sertoli cells were cultured in vitro, exposed to daidzein at 0.03, 0.3, 3, and 30 µmol/L and genistein at 0.05, 0.5, 5 and 50 µmol/L, respectively. MTT was used to detect the proliferation of sertoli cells. Real-time PCR was used to detect the relative mRNA expressions of FSHR, INHα, INHßB, ABP, Tf and vimentin. RESULTS: Compared with control groups, cell proliferation and the relative mRNA expression levels of INHßB and ABP in the treated cells showed no significant alternation. The INHα mRNA expression levels were increased in 0.3 and 3 µmol/L Dai and 0.05 µmol/L Gen, while the mRNA expression levels of FSHR were downregulated in 30 µmol/L Dai and Gen at all concentrations. Tf mRNA expression levels were downregulated in 30 µmol/L Dai and 5 µmol/L and 50 µmol/L Gen, and the mRNA expression levels of vimentin were downregulated in 3 and 30 µmol/L Dai and 50 µmol/L Gen. CONCLUSION: Soy Isoflavones may have potential detrimental effect on the male reproductive system, as they may impact the function of sertoli cells by downregulating the transcription levels of some important proteins.


Assuntos
Isoflavonas/efeitos adversos , Células de Sertoli/efeitos dos fármacos , Testículo/efeitos dos fármacos , Proteína de Ligação a Androgênios/metabolismo , Animais , Subunidades beta de Inibinas/metabolismo , Inibinas/metabolismo , Masculino , RNA Mensageiro , Ratos , Ratos Sprague-Dawley , Receptores do FSH/metabolismo , Glycine max/química , Testículo/citologia , Transferrina/metabolismo
16.
Chin J Integr Med ; 20(3): 194-9, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24615212

RESUMO

OBJECTIVE: To evaluate the effects of the Chinese herbal formula Wuzi Yanzong Pill (, WYP) on the spermatogenesis and specific secretory functions of Sertoli cells in rat model and to investigate the underlying mechanism. METHODS: Five groups of male Sprague-Dawley rats including the control group, the model group, the low-dose WYP group, the medium-dose WYP group and the high-dose WYP group (5 in each group) were treated daily with vehicle, multiglycosides of Tripterygium wilfordii Hook f (GTW) either alone (20 mg/kg) or followed by WYP (0.5, 1.0, or 2.0 g/kg daily), respectively for 30 days. Serum levels of follicle-stimulating hormone (FSH), inhibin B (INHB) and testosterone (T) were evaluated using enzyme-linked immunosorbent assay. Androgen-binding protein (ABP) gene expression and transferrin (TF) protein expression in testis tissue specimens of all rats were determined using real-time reverse transcriptase polymerase chain reaction and Western blotting analysis, respectively. Histopathological alterations in the testis were determined using Johnsen's score. RESULTS: The toxicity of GTW towards Sertoli cell secretory functions and spermatogenesis was accompanied by increased serum FSH concentrations and decreased INHB and T concentrations. Upregulated ABP mRNA levels, and decreased TF protein expression and Johnsen's scores were detected in the model group compared with the control group P<0.05 or P<0.01). Oral high-dose WYP administrations to GTW-treated rats effectively alleviated all of the GTW-induced changes in specific secretory functions of Sertoli cells (ABP, INHB and TF). Furthermore, serum T level and Johnsen's score of the testis increased greatly compared with the model group (P<0.01). CONCLUSION: WYP has the ability to improve the spermatogenesis, possibly through modulating the secretory proteins expression of Sertoli cells.


Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Células de Sertoli/metabolismo , Espermatogênese/efeitos dos fármacos , Proteína de Ligação a Androgênios/genética , Proteína de Ligação a Androgênios/metabolismo , Animais , Western Blotting , Hormônio Foliculoestimulante/sangue , Regulação da Expressão Gênica/efeitos dos fármacos , Inibinas/sangue , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Células de Sertoli/efeitos dos fármacos , Comprimidos , Testículo/citologia , Testículo/metabolismo , Testosterona/sangue , Transferrina/metabolismo
17.
Environ Toxicol Pharmacol ; 37(2): 608-16, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24583408

RESUMO

The aim of this study was to investigate the effects of nonylphenol (NP) exposure on the expression of cell receptors and secretory function in mouse Sertoli TM4 cells. There were no significant changes in mRNA expression of estrogen receptor (ER)-α and toll like receptor (TLR)-4 in the cells exposed to NP for 24h. However, the mRNA expression levels of ER-ß, progesterone receptor (PR) and androgen receptor (AR) were down-regulated in NP groups. Furthermore, NP treatment evoked significant changes in protein expression levels of ER-ß and follicle-stimulating hormone receptor (FSHR). There were significant changes in the mRNA expression levels of vinculin, N-cadherin and occludin, but not vimentin. Levels of inhibin B, androgen binding protein (ABP) and transferrin (Trf) were found to change significantly in NP challenged cells. Additionally, the decrease of nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) mRNA expression and increase of cytokine levels were simultaneously found in NP stimulated TM4 cells. In conclusion, these findings have shown that NP exposure affected expression of cell receptors and may damage specific secretory function of Sertoli TM4 cells, which may be associated with the male-specific reproductive toxicity of NP.


Assuntos
Fenóis/toxicidade , Células de Sertoli/efeitos dos fármacos , Proteína de Ligação a Androgênios/metabolismo , Animais , Hormônio Antimülleriano/genética , Caderinas/genética , Linhagem Celular , Citocinas/metabolismo , Inibinas/metabolismo , Masculino , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Ocludina/genética , RNA Mensageiro/metabolismo , Receptores do FSH/metabolismo , Receptores de Esteroides/genética , Células de Sertoli/metabolismo , Receptor 4 Toll-Like/genética , Transferrina/genética , Vimentina/genética , Vinculina/genética
18.
Mol Endocrinol ; 28(2): 239-48, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24422631

RESUMO

The androgen receptor (AR) plays a pivotal role in prostate homeostasis and prostate cancer development. To understand the mechanism underlying the regulation of the AR holds a promise for developing novel therapeutic approaches for prostate cancer. Here, we show that the Von Hippel-Lindau gene product, pVHL, physically interacts with AR and inhibits AR transcription activity but does not induce AR turnover. Moreover, pVHL also suppresses androgen-induced cell proliferation, implicating a physiological role of pVHL in androgen-induced signaling pathway. In addition, we provide evidence to show that pVHL actually enhanced AR de-ubiquitination instead of inducing AR ubiquitination, uncovering a noncanonical role of pVHL in the ubiquitin proteasome pathway. Our data reveal a novel function of pVHL in the regulation of AR transcription activity, which may expand the scope of pVHL in tumor suppression and provide mechanistic insight into prostate cancer initiation and progression.


Assuntos
Receptores Androgênicos/metabolismo , Ubiquitinação , Proteína Supressora de Tumor Von Hippel-Lindau/fisiologia , Proteína de Ligação a Androgênios/genética , Proteína de Ligação a Androgênios/metabolismo , Animais , Linhagem Celular Tumoral , Proliferação de Células , Proteínas de Ligação a DNA/metabolismo , Expressão Gênica , Células HEK293 , Humanos , Calicreínas/genética , Calicreínas/metabolismo , Regiões Promotoras Genéticas , Antígeno Prostático Específico/genética , Antígeno Prostático Específico/metabolismo , Domínios e Motivos de Interação entre Proteínas , Mapeamento de Interação de Proteínas , Ratos , Receptores Androgênicos/química , Serina Endopeptidases/genética , Serina Endopeptidases/metabolismo , Ativação Transcricional , Proteína Supressora de Tumor Von Hippel-Lindau/química
19.
Andrologia ; 46(8): 902-9, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24079456

RESUMO

Testicular steroidogenesis has significant implication in male reproductive function. Although the effects of various signalling molecules on testicular functions have been studied earlier, the influence of the plant hormone gibberellic acid (GA3 ) on steroidogenesis has not been investigated. Acute (4 h) and subacute (15 days) studies using this compound through oral administration (150 µg day(-1) ) to groups of normal and diabetic Wistar male rats were therefore carried out. Results indicate that (i) enhanced activity of steroidogenic markers 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase (17ß-HSD), elevated tissue testosterone (T) content, increased steroidogenic acute regulatory protein (StAR) and androgen binding protein (ABP) levels with reduced lipid peroxidation and improved antioxidant defence in this treatment group of normal and diabetic rat testis, and (ii) elevated lipid peroxidation and diminished antioxidant defence, with insignificant change in 3ß-HSD and 17ß-HSD activity and testosterone level in acute treatment group of normal and diabetic rats testis, were noted. The observed increase in the activity of testicular 3ß-HSD and 17ß-HSD along with elevated testosterone content established GA3 as an inducer of steroidogenesis in rat.


Assuntos
Giberelinas/farmacologia , Hormônios Esteroides Gonadais/biossíntese , Reguladores de Crescimento de Plantas/farmacologia , Testículo/efeitos dos fármacos , 17-Hidroxiesteroide Desidrogenases/metabolismo , 3-Hidroxiesteroide Desidrogenases/metabolismo , Proteína de Ligação a Androgênios/metabolismo , Animais , Antioxidantes/metabolismo , Diabetes Mellitus Experimental/metabolismo , Avaliação Pré-Clínica de Medicamentos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Fosfoproteínas/metabolismo , Ratos Wistar , Testículo/metabolismo
20.
Nan Fang Yi Ke Da Xue Xue Bao ; 33(11): 1709-13, 2013 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-24273285

RESUMO

OBJECTIVE: To investigate the effect of different doses of glyphosate on apoptosis and expressions of androgen-binding protein (ABP) and vimentin mRNA in mouse Sertoli cells. METHODS: Primarily cultured mouse Sertoli cells incubated with different doses of glyphosate (60, 90, 120, 150 and 180 mg/L) for 24 h. The growth and morphological alterations in the cells were observed under inverted microscope, and the cell proliferation rate was evaluated withMTT assay. Hoechst 33342 staining was used to detect cell apoptosis after the treatment, and RT-PCR was performed to examine the changes in the expression of ABP and vimentin mRNAs. RESULTS: Sertoli cells exposed to glyphosate showed a reduced cell volume, cell dissociation with occasional cell disruption. The proliferation of the exposed was suppressed with an increased rate of cell apoptosis and lowered expressions of ABP and vimentin mRNAs (P<0.05). CONCLUSION: GLY can cause cellular damages, inhibit cell proliferation, induce cell apoptosis, and decrease expression of ABP and vimentin mRNAs in mouse Sertoli cells in vitro.


Assuntos
Proteína de Ligação a Androgênios/genética , Apoptose/efeitos dos fármacos , Glicina/análogos & derivados , Herbicidas/toxicidade , Células de Sertoli/efeitos dos fármacos , Vimentina/genética , Proteína de Ligação a Androgênios/metabolismo , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Glicina/administração & dosagem , Glicina/toxicidade , Herbicidas/administração & dosagem , Masculino , Camundongos , RNA Mensageiro/metabolismo , Células de Sertoli/citologia , Células de Sertoli/metabolismo , Vimentina/metabolismo , Glifosato
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