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1.
Nat Commun ; 10(1): 4344, 2019 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-31554802

RESUMO

Innate immune responses to Zika virus (ZIKV) are dampened in the lower female reproductive tract (LFRT) compared to other tissues, but the mechanism that underlies this vulnerability is poorly understood. Using tissues from uninfected and vaginally ZIKV-infected macaques and mice, we show that low basal expression of RNA-sensing pattern recognition receptors (PRRs), or their co-receptors, in the LFRT contributes to high viral replication in this tissue. In the LFRT, ZIKV sensing provides limited protection against viral replication, and the sensors are also minimally induced after vaginal infection. While IFNα/ß receptor signaling offers minimal protection in the LFRT, it is required to prevent dissemination of ZIKV to other tissues, including the upper FRT. Our findings support a role for RNA-sensing PRRs in the dampened innate immunity against ZIKV in the LFRT compared to other tissues and underlie potential implications for systemic dissemination upon heterosexual transmission of ZIKV in women.


Assuntos
Genitália Feminina/imunologia , Imunidade Inata/imunologia , RNA Viral/imunologia , Infecção por Zika virus/imunologia , Zika virus/imunologia , Animais , Feminino , Regulação Viral da Expressão Gênica , Genitália Feminina/metabolismo , Genitália Feminina/virologia , Humanos , Imunidade Inata/genética , Macaca mulatta , Camundongos Endogâmicos C57BL , Camundongos Knockout , RNA Viral/genética , Receptor de Interferon alfa e beta/genética , Receptor de Interferon alfa e beta/imunologia , Receptor de Interferon alfa e beta/metabolismo , Receptores de Reconhecimento de Padrão/genética , Receptores de Reconhecimento de Padrão/imunologia , Receptores de Reconhecimento de Padrão/metabolismo , Receptor 3 Toll-Like/genética , Receptor 3 Toll-Like/imunologia , Receptor 3 Toll-Like/metabolismo , Vagina/imunologia , Vagina/metabolismo , Vagina/virologia , Replicação Viral/genética , Replicação Viral/imunologia , Zika virus/genética , Zika virus/fisiologia , Infecção por Zika virus/genética , Infecção por Zika virus/virologia
2.
Future Microbiol ; 14: 839-846, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31373210

RESUMO

Aim: The primary objective of this study was to evaluate the effects of polypeptide-enriched Gastrodia elata extracts (GE) on vulvovaginal candidiasis (VVC). Materials & methods: A VVC model induced by Candida albicans (C. albicans) infection was successfully developed in BALB/c mice. After treatment, the colony-forming unit (CFU) of vaginal lavage was measured by plating. The extent of the inflammatory response was assessed by hematoxylin-eosin (H&E) staining and enzyme-linked immunosorbent assay (ELISA). Results: GE had an inhibitory effect on the proliferation of C. albicans and inflammatory reaction. Meanwhile, it had a potentially beneficial effect on the growth of Lactobacillus. Conclusion: These results showed the potential application of GE as an antifungal agent in VVC treatment.


Assuntos
Candida albicans/efeitos dos fármacos , Candidíase Vulvovaginal/tratamento farmacológico , Gastrodia/química , Peptídeos/farmacologia , Peptídeos/uso terapêutico , Animais , Antifúngicos/química , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candida albicans/crescimento & desenvolvimento , Candidíase Vulvovaginal/sangue , Candidíase Vulvovaginal/microbiologia , Candidíase Vulvovaginal/patologia , Modelos Animais de Doenças , Feminino , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Inflamação/patologia , Interleucinas/sangue , Interleucinas/metabolismo , Camundongos Endogâmicos BALB C , Peptídeos/química , Resultado do Tratamento , Vagina/metabolismo , Vagina/microbiologia , Vagina/patologia
3.
Biomed Res Int ; 2019: 8921284, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31467917

RESUMO

Ideal animal models are needed to reflect the changes in the biochemical and biomechanical properties of the vagina that occur in pelvic organ prolapse (POP). In this study, we aimed to demonstrate the short and long-term effect of menopause on the biochemical and biomechanical properties of rat anterior vaginas. Here, Sprague-Dawley rats were bilaterally ovariectomized to induce menopause. Rats without ovariectomy served as the normal control group (n=12). The histology changes and the expression of collagen I, III, and a-SMA were assessed to indicate the biochemical changes in the vagina 2 weeks, 4 weeks, and 16 weeks after ovariectomy (n=6 for 2 and 4 weeks, n=12 for 16 weeks). Uniaxial biomechanical testing was conducted in the control group and ovariectomized rats 16 weeks after ovariectomy. Compared with the control group, the ovariectomy group showed a significant increase in the expression of collagen I 2 weeks after ovariectomy, while collagen III showed a declining trend. Two weeks after ovariectomy, the smooth muscle bundles began to become disorganized, and the fraction of smooth muscle in the nonvascular muscularis of the proximal vagina was significantly decreased (P<0.001). However, there was no difference in the expression of a-SMA in the distal vagina. Compared with the control group, the ovariectomy group had stiffer vaginas with a declining trend in the ultimate load 16 weeks after ovariectomy. In conclusion, surgically induced menopause had a significant short- and long-term impact on tissue composition and biomechanical properties of the rat vagina, which may lead to increased susceptibility to POP development.


Assuntos
Colágeno/metabolismo , Ovariectomia , Vagina/metabolismo , Animais , Colágeno/genética , Feminino , Humanos , Menopausa/genética , Menopausa/metabolismo , Músculo Liso/metabolismo , Prolapso de Órgão Pélvico/metabolismo , Ratos , Ratos Sprague-Dawley , Vagina/fisiologia
4.
Gene ; 711: 143937, 2019 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-31228541

RESUMO

BACKGROUND & OBJECTIVES: Vaginal atrophy is characterized by thinning of vaginal epithelial layers and decreased local blood flow. We aimed to evaluate the regenerative effects of Adipose derived mesenchymal stem cells (ADMSC) and Bone marrow derived mesenchymal stem cells (BMDSC) on vaginal atrophy in rat menopause model. MATERIALS AND METHODS: Rats were randomly divided into 4 (four) groups: sham, control, ADMSC, BMDSC. Vaginal epithelial thickness, structure of the lamina propria, blood vessels in the lamina propria, collagen deposition, and muscle structure were evaluated. Anti ER α, VEGF, VEGFR 1, Bax and bcl-2 antibodies were analyzed. Beta actin gene was used as endogenous control. Genetical differences among the groups were compared by using Kruskal Wallis and Mann Whitney U test. p < 0.05 was regarded as statistically significant. RESULTS: Epithelial thickness of ADMSC group was higher than control group, but less than sham group Epithelial thickness of BMDSC group was less than sham group. Lamina propria and muscle tissue of ADMSC and BMDSC groups were found to be similar to sham group. VEGFR-1, VEGF, Bax and ER-α staining levels were higher in ADMSC and BMDSC groups than control group. ADMSC group stained stronger with VEGFR-1 and VEGF than BMDSC group. Bcl-2 staining level was increased in ADMSC applied group. No statistically significant difference was detected in Bax and Bcl-2 genes and Bax-/Bcl-2 ratio. CONCLUSIONS: Although genetic expression might have ended and could not be significantly demonstrated, histological and immunohistochemical results favor ADMSC application in vaginal atrophy rather than BMDSC.


Assuntos
Tecido Adiposo/citologia , Biomarcadores/metabolismo , Células da Medula Óssea/citologia , Menopausa/fisiologia , Transplante de Células-Tronco Mesenquimais/métodos , Vagina/patologia , Tecido Adiposo/metabolismo , Animais , Atrofia , Células da Medula Óssea/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Receptor alfa de Estrogênio/metabolismo , Feminino , Humanos , Menopausa/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Ratos , Vagina/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Proteína X Associada a bcl-2/metabolismo
5.
PLoS One ; 14(5): e0217229, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31107913

RESUMO

Recent data support that the vaginal microbiota may alter mucosal pharmacokinetics (PK) of topically delivered microbicides. Our team developed an intravaginal ring (IVR) that delivers tenofovir (TFV) (8-10 mg/day) alone or with levonorgestrel (LNG) (20 ug/day). We evaluated the effect of IVRs on the vaginal microbiota, and describe how the vaginal microbiota impacts mucosal PK of TFV. CONRAD A13-128 was a randomized, placebo controlled phase I study. We randomized 51 women to TFV, TFV/LNG or placebo IVR. We assessed the vaginal microbiota by sequencing the V3-V4 regions of 16S rRNA genes prior to IVR insertion and after approximately 15 days of use. We measured the concentration of TFV in the cervicovaginal (CV) aspirate, and TFV and TFV-diphosphate (TFV-DP) in vaginal tissue at the end of IVR use. The change in relative or absolute abundance of vaginal bacterial phylotypes was similar among active and placebo IVR users (all q values >0.13). TFV concentrations in CV aspirate and vaginal tissue, and TFV-DP concentrations in vaginal tissue were not significantly different among users with community state type (CST) 4 versus those with Lactobacillus dominated microbiota (all p values >0.07). The proportions of participants with CV aspirate concentrations of TFV >200,000 ng/mL and those with tissue TFV-DP concentrations >1,000 fmol/mg were similar among women with anaerobe versus Lactobacillus dominated microbiota (p = 0.43, 0.95 respectively). There were no significant correlations between the CV aspirate concentration of TFV and the relative abundances of Gardnerella vaginalis or Prevotella species. Tissue concentrations of TFV-DP did not correlate with any the relative abundances of any species, including Gardnerella vaginalis. In conclusion, active IVRs did not differ from the placebo IVR on the effect on the vaginal microbiota. Local TFV and TFV-DP concentrations were high and similar among IVR users with Lactobacillus dominated microbiota versus CST IV vaginal microbiota. Trial registration: ClinicalTrials.gov NCT02235662.


Assuntos
Fármacos Anti-HIV/administração & dosagem , Fármacos Anti-HIV/farmacocinética , Dispositivos Anticoncepcionais Femininos , Levanogestrel/administração & dosagem , Microbiota/efeitos dos fármacos , Tenofovir/administração & dosagem , Tenofovir/farmacocinética , Vagina/metabolismo , Vagina/microbiologia , Adenina/análogos & derivados , Adenina/farmacocinética , Adulto , Antivirais/administração & dosagem , Antivirais/farmacocinética , Remoção de Dispositivo , Feminino , Infecções por HIV/prevenção & controle , Herpes Genital/prevenção & controle , Humanos , Microbiota/genética , Pessoa de Meia-Idade , Membrana Mucosa/efeitos dos fármacos , Membrana Mucosa/metabolismo , Organofosfatos/farmacocinética , Vagina/efeitos dos fármacos , Adulto Jovem
6.
Mol Med Rep ; 19(6): 4727-4734, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31059065

RESUMO

The aim of the present study was to investigate the protective effect of integrin ß1 in the treatment of stress urinary incontinence (SUI) by electrical stimulation, and the underlying mechanisms by which electrical stimulation regulates the collagen metabolism of female vaginal wall fibroblasts (FVWFs). FVWFs obtained from the vaginal wall tissue of patients with (Ingelman­Sundberg scale; grade II, n=8; grade III, n=10) or without (n=8) SUI during gynecological operations were isolated by enzymatic digestion and subsequently identified by immunocytochemistry. Following this, cultured FVWFs were treated with an inhibitor of integrin ß1, recombinant human integrin ß1 and electrical stimulation (100 mv/mm, 2 h, 20 Hz), followed by total mRNA and protein extraction. mRNA and protein expression levels of integrin ß1, transforming growth factor (TGF)­ß1 and collagen (COL) I and III in FVWFs were quantified by reverse transcription­quantitative PCR (RT­qPCR) and western blot analysis respectively. Integrin ß1, TGF­ß1 and COL I and III expression levels were decreased in patients with SUI compared with healthy controls, and the grade III group had lower levels than the grade II group. Following electrical stimulation treatment, the expression levels of TGF­ß1, COL I and III were enhanced in the grade II group, but not in the grade III group. Nevertheless, the inhibitor of integrin ß1 reduced the protective effect of electrical stimulation in the grade II group. In addition, electrical stimulation combined with recombinant human integrin ß1 could also protect cells from SUI in the grade III group. The present study provides evidence for the increased degradation of the extracellular matrix and integrin ß1 in the vaginal wall tissues of patients with SUI, and the protective effect of electrical stimulation against SUI via integrin ß1. These results provide a novel mechanism for the treatment of SUI using electrical stimulation.


Assuntos
Estimulação Elétrica/métodos , Integrina beta1/farmacologia , Integrina beta1/uso terapêutico , Incontinência Urinária por Estresse/tratamento farmacológico , Colágeno Tipo I/metabolismo , Colágeno Tipo III/metabolismo , Matriz Extracelular/metabolismo , Feminino , Fibroblastos/metabolismo , Fibroblastos/patologia , Humanos , Fator de Crescimento Transformador beta1 , Incontinência Urinária , Vagina/metabolismo , Vagina/patologia
7.
J Nanobiotechnology ; 17(1): 65, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-31092246

RESUMO

It is essential that prophylactic drugs do not interfere with the normal function of the immune system. The use of nanoparticles as vaginal microbicides is a promising prevention strategy against sexually transmitted infections. With that aim, our group is working with the G2-S16, a second generation carbosilane dendrimer with sulfonate groups in the periphery, which has been previously shown to be effective against HIV-1 and HSV-2 infection, and it is now on the road to clinical trials. Our objective in this new study is to assess the effects of G2-S16 on the immune barrier of the female reproductive tract. The expression of differentiation, maturation and activation markers was measured in epithelial cells, dendritic cells, M and GM macrophages, and T cells using RT-qPCR and flow cytometry. The results demonstrate that G2-S16 does not alter the natural immunity of the vagina, strongly supporting the biosafety of this dendrimer for clinical use.


Assuntos
Anti-Infecciosos/farmacologia , Dendrímeros/química , Sistema Imunitário/efeitos dos fármacos , Silanos/química , Vagina/metabolismo , Anti-Infecciosos/administração & dosagem , Anti-Infecciosos/efeitos adversos , Biomarcadores/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Dendríticas/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Macrófagos/efeitos dos fármacos , Tamanho da Partícula , RNA Mensageiro/metabolismo , Transdução de Sinais , Linfócitos T/efeitos dos fármacos
8.
J Mass Spectrom ; 54(8): 693-703, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31116903

RESUMO

Cervicovaginal fluid (CVF) is a valuable source of clinical information about the female reproductive tract in both nonpregnant and pregnant women. The aim of this study is to specify the CVF proteome at different stages of cervix neoplastic transformation by label-free quantitation approach based on liquid chromatography tandem mass spectrometry (LC-MS/MS) method. The proteome composition of CVF from 40 women of reproductive age with human papillomavirus (HPV)-associated cervix neoplastic transformation (low-grade squamous intraepithelial lesion [LSIL], high-grade squamous intraepithelial lesion [HSIL], and CANCER) was investigated. Hierarchical clustering and principal component analysis (PCA) of the proteomic data obtained by a label-free quantitation approach show the distribution of the sample set between four major clusters (no intraepithelial lesion or malignancy [NILM], LSIL, HSIL and CANCER) depending on the form of cervical lesion. Multisample ANOVA with subsequent Welch's t test resulted in 117 that changed significantly across the four clinical stages, including 27 proteins significantly changed in cervical cancer. Some of them were indicated as promising biomarkers previously (ACTN4, VTN, ANXA1, CAP1, ANXA2, and MUC5B). CVF proteomic data from the discovery stage were analyzed by the partial least squares-discriminant analysis (PLS-DA) method to build a statistical model, allowing to differentiate severe dysplasia (HSIL and CANCER) from the mild/normal stage (NILM and LSIL), and receiver operating characteristic (ROC) area under the curve (AUC) were obtained on an independent set of 33 samples. The sensitivity of the model was 77%, and the specificity was 94%; AUC was equal to 0.87. CVF proteome proved to be reflect the stage of cervical epithelium neoplastic process.


Assuntos
Líquidos Corporais/metabolismo , Transformação Celular Neoplásica/metabolismo , Colo do Útero/metabolismo , Proteoma/análise , Neoplasias do Colo do Útero/diagnóstico , Vagina/metabolismo , Adulto , Biomarcadores/análise , Transformação Celular Neoplásica/patologia , Colo do Útero/patologia , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Papillomaviridae/fisiologia , Infecções por Papillomavirus/metabolismo , Gravidez , Proteoma/metabolismo , Espectrometria de Massas em Tandem , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologia , Vagina/patologia , Esfregaço Vaginal
9.
J Appl Microbiol ; 127(2): 565-575, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31102489

RESUMO

AIMS: To observe the therapeutic effects of vaginal infusion of probiotic Clostridium butyricum WZ001 on bacterial vaginosis (BV) in mice. METHODS AND RESULTS: Female ICR mice were used to establish the model of BV by infecting oestrogen-treated mice with Escherichia coli, and then treated with high- and low dose of C. butyricum. Clinical indexes of mice in the C. butyricum-treated groups were significantly improved and comparable to those in the antibiotic group. Pap staining showed that neutrophil count was significantly increased after modelling and largely decreased after C. butyricum treatment (P < 0·01). Dynamic observation of E. coli and Lactobacillus showed that the number of E. coli significantly decreased in the C. butyricum-treated groups or in the antibiotic group with prolonged treatment (P < 0·01). Besides, the number of E. coli in the low-dose C. butyricum group was higher than that in either its high-dose counterpart or the antibiotic group respectively (P < 0·01). The number of Lactobacillus decreased evidently in the antibiotic group (P < 0·01), while that in the C. butyricum groups remained consistent. Moreover, C. butyricum inhibited the proliferation of E. coli by the experiment in vitro. The phosphorylation of nuclear factor-kappa B (NF-κB) p65 in vaginal tissue and the serum levels of inflammatory cytokines, IL-1ß, TNF-α and IL-6, increased after modelling and significantly decreased after treated with C. butyricum (P < 0·01), with no difference found when compared with the antibiotic group. CONCLUSION: Clostridium butyricum inhibits the growth of pathogenic bacteria as well as the inflammatory response induced by E. coli and promotes the growth of Lactobacillus to maintain the vaginal micro-ecological balance. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggest that probiobitc C. butyricum WZ001 has a great potential in the clinical treatment of BV.


Assuntos
Clostridium butyricum , Infecções por Escherichia coli/terapia , Probióticos/uso terapêutico , Vaginose Bacteriana/terapia , Animais , Citocinas/sangue , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/metabolismo , Infecções por Escherichia coli/microbiologia , Feminino , Lactobacillus/isolamento & purificação , Camundongos , Camundongos Endogâmicos ICR , Fator de Transcrição RelA/metabolismo , Vagina/metabolismo , Vagina/microbiologia , Vaginose Bacteriana/imunologia , Vaginose Bacteriana/metabolismo , Vaginose Bacteriana/microbiologia
10.
Int J Pharm ; 564: 207-213, 2019 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-30999049

RESUMO

The past fifteen years have witnessed a resurgence of interest in vaginal ring technologies for drug delivery applications, mostly driven by the impetus for development of vaginally-administered antiretroviral microbicides to help reduce the high acquisition rates for human immunodeficiency virus (HIV) among Sub-Saharan African women. Currently, the lead candidate microbicide is a 28-day silicone elastomer vaginal ring releasing dapivirine (Ring-004), an experimental non-nucleoside reverse transcriptase inhibitor. The ring was tested in two pivotal Phase III clinical studies in 2016 and is currently undergoing review by the European Medicines Agency. Recently, we described a new type of silicone elastomer vaginal ring offering sustained release of the protein molecule 5P12-RANTES, a potent experimental chemokine analogue that potently blocks the HIV CCR5 coreceptor. Building on our previous work, here we report the preclinical development of a new combination vaginal ring that offers sustained release of both 5P12-RANTES and dapivirine, in which the 5P12-RANTES is incorporated into an exposed core within the ring body and the dapivirine in the sheath. In this way, in vitro release of dapivirine matches closely that for Ring-004. Also, we report the pharmacokinetic testing of this combination ring formulation in sheep, where vaginal concentrations of both drugs are maintained over 28 days at levels potentially useful for preventing HIV infection in women.


Assuntos
Fármacos Anti-HIV/administração & dosagem , Quimiocinas CC/administração & dosagem , Dispositivos Anticoncepcionais Femininos , Infecções por HIV/prevenção & controle , Pirimidinas/administração & dosagem , Animais , Fármacos Anti-HIV/farmacocinética , Quimiocinas CC/farmacocinética , Preparações de Ação Retardada/administração & dosagem , Combinação de Medicamentos , Feminino , Pirimidinas/farmacocinética , Ovinos , Vagina/metabolismo
11.
Pharm Biol ; 57(1): 226-230, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30946631

RESUMO

CONTEXT: We identified an active prenylated derivative of genistein, 8-prenylgenistein (8PG) from Erythrina variegata L. (Leguminosae) and found that 8PG increased osteoprotective effects of genistein in oestrogen-deficient mice. OBJECTIVE: This study investigated and compared the oestrogenic effects of genistein and 8PG on uterus and vagina of immature mice. MATERIALS AND METHODS: Immature female CD-1 mice were orally treated with vehicle (Control, n = 10) or genistein (75 mg/kg, n = 10) or 8PG with low (8PG-L, 75 mg/kg, n = 10) and high dose (8PG-H, 150 mg/kg, n = 10) for 7 consecutive days by intragastric gavage. The uterus and vagina were harvested for histological and molecular measurements. RESULTS: Treatment with genistein and 8PG-H significantly increased uterus index (1.98 ± 0.21 & 1.49 ± 0.16 mg/g) and vagina index (3.83 ± 0.11 & 3.13 ± 0.25 mg/g) as compared to untreated control (uterus, 1.12 ± 0.13 mg/g; vagina, 2.32 ± 0.18 mg/g). Accordingly, both genistein and 8PG-H made vaginal cells keratinized and induced uterine and vaginal hypertrophy associated with the endometrial proliferation. 8PG-L did not affect oestrus cycle and histology of uterus and vagina. Treatment of immature mice with genistein or 8PG-H upregulated protein expression of oestrogen receptor-α (ER-α) and proliferating cell nuclear antigen (PCNA), but 8PG-L did not alter ER-α and PCNA expression in uterus and vagina. CONCLUSION: This study indicated that 8-prenylgenistein exerted oestrogenic effects in immature female mice. The efficacy and safety of 8-prenylgenistein when applied in improving oestrogen deficiency-induced syndrome requires further elucidation.


Assuntos
Estrogênios/farmacologia , Genisteína/análogos & derivados , Genisteína/farmacocinética , Útero/efeitos dos fármacos , Vagina/efeitos dos fármacos , Administração Oral , Animais , Relação Dose-Resposta a Droga , Receptor alfa de Estrogênio/metabolismo , Estrogênios/administração & dosagem , Estrogênios/toxicidade , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Genisteína/administração & dosagem , Genisteína/farmacologia , Genisteína/toxicidade , Camundongos , Regulação para Cima/efeitos dos fármacos , Útero/metabolismo , Vagina/metabolismo
12.
EBioMedicine ; 44: 675-690, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31027917

RESUMO

BACKGROUND: Dysbiotic vaginal microbiota have been implicated as contributors to persistent HPV-mediated cervical carcinogenesis and genital inflammation with mechanisms unknown. Given that cancer is a metabolic disease, metabolic profiling of the cervicovaginal microenvironment has the potential to reveal the functional interplay between the host and microbes in HPV persistence and progression to cancer. METHODS: Our study design included HPV-negative/positive controls, women with low-grade and high-grade cervical dysplasia, or cervical cancer (n = 78). Metabolic fingerprints were profiled using liquid chromatography-mass spectrometry. Vaginal microbiota and genital inflammation were analysed using 16S rRNA gene sequencing and immunoassays, respectively. We used an integrative bioinformatic pipeline to reveal host and microbe contributions to the metabolome and to comprehensively assess the link between HPV, microbiota, inflammation and cervical disease. FINDINGS: Metabolic analysis yielded 475 metabolites with known identities. Unique metabolic fingerprints discriminated patient groups from healthy controls. Three-hydroxybutyrate, eicosenoate, and oleate/vaccenate discriminated (with excellent capacity) between cancer patients versus the healthy participants. Sphingolipids, plasmalogens, and linoleate positively correlated with genital inflammation. Non-Lactobacillus dominant communities, particularly in high-grade dysplasia, perturbed amino acid and nucleotide metabolisms. Adenosine and cytosine correlated positively with Lactobacillus abundance and negatively with genital inflammation. Glycochenodeoxycholate and carnitine metabolisms connected non-Lactobacillus dominance to genital inflammation. INTERPRETATION: Cervicovaginal metabolic profiles were driven by cancer followed by genital inflammation, HPV infection, and vaginal microbiota. This study provides evidence for metabolite-driven complex host-microbe interactions as hallmarks of cervical cancer with future translational potential. FUND: Flinn Foundation (#1974), Banner Foundation Obstetrics/Gynecology, and NIH NCI (P30-CA023074).


Assuntos
Metaboloma , Microbiota , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/virologia , Neoplasias do Colo do Útero/etiologia , Neoplasias do Colo do Útero/metabolismo , Vaginite/etiologia , Vaginite/metabolismo , Adulto , Aminoácidos/metabolismo , Biologia Computacional/métodos , Suscetibilidade a Doenças , Feminino , Humanos , Metabolismo dos Lipídeos , Metabolômica/métodos , RNA Ribossômico 16S/genética , Curva ROC , Neoplasias do Colo do Útero/patologia , Vagina/metabolismo , Vagina/microbiologia , Vagina/patologia , Vagina/virologia , Xenobióticos/metabolismo
13.
Arch Biochem Biophys ; 666: 127-137, 2019 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-30914253

RESUMO

Female reproductive tissues undergo significant alterations during pregnancy, which may compromise the structural integrity of extracellular matrix proteins. Here, we report on modifications of elastic fibers, which are primarily composed of elastin and believed to provide a scaffold to the reproductive tissues, due to parity and parturition. Elastic fibers from the upper vaginal wall of virgin Sprague Dawley rats were investigated and compared to rats having undergone one, three, or more than five pregnancies. Optical microscopy was used to study fiber level changes. Mass spectrometry, 13C and 2H NMR, was applied to study alterations of elastin from the uterine horns. Spectrophotometry was used to measure matrix metalloproteinases-2,9 and tissue inhibitor of metalloproteinase-1 concentration changes in the uterine horns. Elastic fibers were found to exhibit increase in tortuosity and fragmentation with increased pregnancies. Surprisingly, secondary structure, dynamics, and crosslinking of elastin from multiparous cohorts appear similar to healthy mammalian tissues, despite fragmentation observed at the fiber level. In contrast, elastic fibers from virgin and single pregnancy cohorts are less fragmented and comprised of elastin exhibiting structure and dynamics distinguishable from multiparous groups, with reduced crosslinking. These alterations were correlated to matrix metalloproteinases-2,9 and tissue inhibitor of metalloproteinase-1 concentrations. This work indicates that fiber level alterations resulting from pregnancy and/or parturition, such as fragmentation, rather than secondary structure (e.g. elastin crosslinking density), appear to govern scaffolding characteristics in the female reproductive tissues.


Assuntos
Elastina/química , Paridade/fisiologia , Vagina/metabolismo , Animais , Desmosina/metabolismo , Tecido Elástico/química , Tecido Elástico/metabolismo , Elastina/metabolismo , Feminino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Ressonância Magnética Nuclear Biomolecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Gravidez , Estrutura Secundária de Proteína , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Inibidor Tecidual de Metaloproteinase-1/metabolismo
14.
BMC Infect Dis ; 19(1): 218, 2019 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-30832593

RESUMO

BACKGROUND: Chlamydia trachomatis infections in women continue to be a major public health concern due to their high prevalence and consequent reproductive morbidities. While antibiotics are usually efficient to clear the Chlamydia, repeat infections are common and may contribute to pathological outcomes. Interferon-gamma (IFN-γ)-mediated immunity has been suggested to be protective against reinfection, and represent an important anti-chlamydial agent, primarily via the induction of indoleamine-2,3 dioxygenase 1 (IDO1) enzyme. IDO1 catalyzes the degradation of tryptophan, which can eliminate C. trachomatis infection in vitro. Here, we sought to measure IDO1 expression levels and related immune markers during different C. trachomatis infection statuses (repeated vs single infection vs post antibiotic treatment), in vitro and in vivo. METHODS: In this study, we measured the expression levels of IDO1 and immune regulatory markers, transforming growth factor ß1 (TGF-ß1) and forkhead box P3 (FoxP3), in vaginal swab samples of C. trachomatis-infected women, with either single or repeated infection. In addition, we used an in vitro co-culture model of endometrial carcinoma cell-line and peripheral blood mononuclear cells (PBMCs) to measure the same immune markers. RESULTS: We found that in women with repeated C. trachomatis infections vaginal IDO1 and TGF-ß1 expression levels were significantly increased. Whereas, women who cleared their infection post antibiotic treatment, had increased levels of IDO1 and TGF-ß1, as well as FoxP3. Similarly, using the in vitro model, we found significant upregulation of IDO1 and TGF-ß1 levels in the co-culture infected with C. trachomatis. Furthermore, we found that in PBMCs infected with C. trachomatis there was a significant upregulation in IDO1 levels, which was independent of IFN-γ. In fact, C. trachomatis infection in PBMCs failed to induce IFN-γ levels in comparison to the uninfected culture. CONCLUSIONS: Our data provide evidence for a regulatory immune response comprised of IDO1, TGF-ß1 and FoxP3 in women post antibiotic treatment. In this study, we demonstrated a significant increase in IDO1 expression levels in response to C. trachomatis infection, both in vivo and in vitro, without elevated IFN-γ levels. This study implicates IDO1 and TGF-ß1 as part of the immune response to repeated C. trachomatis infections, independently of IFN-γ.


Assuntos
Infecções por Chlamydia/diagnóstico , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Interferon gama/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Adulto , Antibacterianos/uso terapêutico , Linhagem Celular , Infecções por Chlamydia/tratamento farmacológico , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/patogenicidade , Técnicas de Cocultura , Feminino , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Humanos , Indolamina-Pirrol 2,3,-Dioxigenase/genética , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/metabolismo , Recidiva , Fator de Crescimento Transformador beta1/genética , Regulação para Cima , Vagina/metabolismo , Vagina/microbiologia , Adulto Jovem
15.
Recent Pat Drug Deliv Formul ; 13(1): 3-15, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30767755

RESUMO

BACKGROUND: Vaginal drug delivery approach represents one of the imperative strategies for local and systemic delivery of drugs. The peculiar dense vascular networks, mucus permeability, and range of physiological characteristics of the vaginal cavity have been exploited for therapeutic benefit. Furthermore, the vaginal drug delivery has been curtailed due to the influence of different physiological factors like acidic pH, constant cervical secretion, microflora, cyclic changes during periods along with turnover of mucus of varying thickness. OBJECTIVE: This review highlights advancement of nanomedicine and its prospective progress towards the clinic. METHODS: Relevant literature reports and patents related to topics are retrieved and used. RESULT: The extensive literature search and patent revealed that nanocarriers are efficacious over conventional treatment approaches. CONCLUSION: Recently, nanotechnology based drug delivery approach has promised better therapeutic outcomes by providing enhanced permeation and sustained drug release activity. Different nanoplatforms based on drugs, peptides, proteins, antigens, hormones, nucleic material, and microbicides are gaining momentum for vaginal therapeutics.


Assuntos
Portadores de Fármacos/metabolismo , Sistemas de Liberação de Medicamentos/métodos , Vagina/metabolismo , Portadores de Fármacos/administração & dosagem , Sistemas de Liberação de Medicamentos/tendências , Feminino , Humanos , Nanomedicina/métodos , Nanomedicina/tendências , Nanopartículas/administração & dosagem , Nanopartículas/metabolismo , Doenças Sexualmente Transmissíveis/tratamento farmacológico , Doenças Sexualmente Transmissíveis/metabolismo , Vagina/efeitos dos fármacos
16.
J Ethnopharmacol ; 236: 9-20, 2019 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-30771519

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Marantodes pumilum (MP) (Kacip Fatimah) is used to maintain the well-being of post-menopausal women. However, its role in ameliorating post menopause-related vaginal atrophy (VA) is unknown. AIMS: To investigate the ability of intravaginal MP gel treatment to ameliorate VA in sex-steroid deficient condition, mimicking post-menopause. METHODS: Ovariectomized female Sprague-Dawley rats received MP (100 µg/ml, 250 µg/ml and 500 µg/ml) and estriol (E) gels intravaginally for seven consecutive days. Rats were then euthanized and vagina was harvested and subjected for histological and protein expression and distribution analyses. Vaginal ultrastructure was observed by transmission electron microscopy (TEM). RESULTS: Thickness of vaginal epithelium increased with increasing intravaginal MP doses. Additionally, increased in expression and distribution of proliferative protein i.e. PCNA, tight junction protein i.e. occludin, water channel proteins i.e. AQP-1 and AQP-2 and proton extruder protein i.e. V-ATPase A1 were observed in the vagina following intravaginal MP and E gels treatment. Intravaginal MP and E gels also induced desmosome formation and approximation of the intercellular spaces between the vaginal epithelium. CONCLUSIONS: Intravaginal MP was able to ameliorate features associated with VA; thus, it has potential to be used as an agent to treat this condition.


Assuntos
Extratos Vegetais/uso terapêutico , Pós-Menopausa , Primulaceae/química , Vagina/efeitos dos fármacos , Vagina/ultraestrutura , Administração Intravaginal , Animais , Atrofia/prevenção & controle , Feminino , Humanos , Microscopia Eletrônica de Transmissão , Ovariectomia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Folhas de Planta/química , Antígeno Nuclear de Célula em Proliferação/metabolismo , Ratos , Ratos Sprague-Dawley , Vagina/metabolismo
17.
Mol Cell Proteomics ; 18(3): 461-476, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30504243

RESUMO

Women at high risk of HIV infection, including sex workers and those with active genital inflammation, have molecular signatures of immune activation and epithelial barrier remodeling in samples of their genital mucosa. These alterations in the local immunological milieu are likely to impact HIV susceptibility. We here analyze host genital protein signatures in HIV uninfected women, with high frequency of condom use, living in HIV-serodiscordant relationships. Cervicovaginal secretions from women living in HIV-serodiscordant relationships (n = 62) were collected at three time points over 12 months. Women living in HIV-negative seroconcordant relationships (controls, n = 25) were sampled at one time point. All study subjects were examined for demographic parameters associated with susceptibility to HIV infection. The cervicovaginal samples were analyzed using a high-throughput bead-based affinity assay. Proteins involved in epithelial barrier function and inflammation were increased in HIV-serodiscordant women. By combining several methods of analysis, a total of five proteins (CAPG, KLK10, SPRR3, elafin/PI3, CSTB) were consistently associated with this study group. Proteins analyzed using the affinity set-up were further validated by label-free tandem mass spectrometry in a partially overlapping cohort with concordant results. Women living in HIV-serodiscordant relationships thus had elevated levels of proteins involved in epithelial barrier function and inflammation despite low prevalence of sexually transmitted infections and a high frequency of safe sex practices. The identified proteins are important markers to follow during assessment of mucosal HIV susceptibility factors and a high-throughput bead-based affinity set-up could be a suitable method for such evaluation.


Assuntos
Colo do Útero/metabolismo , Infecções por HIV/transmissão , Proteômica/métodos , Doenças Sexualmente Transmissíveis/metabolismo , Vagina/metabolismo , Adulto , Colo do Útero/virologia , Análise por Conglomerados , Proteínas Ricas em Prolina do Estrato Córneo/metabolismo , Cistatina B/metabolismo , Diagnóstico Precoce , Elafina/metabolismo , Feminino , Infecções por HIV/metabolismo , Ensaios de Triagem em Larga Escala , Humanos , Calicreínas/metabolismo , Estudos Longitudinais , Masculino , Proteínas dos Microfilamentos/metabolismo , Proteínas Nucleares/metabolismo , Parceiros Sexuais , Espectrometria de Massas em Tandem , Vagina/virologia , Adulto Jovem
18.
Int J Impot Res ; 31(1): 46-49, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30254307

RESUMO

The purpose of this study was to identify and localize mucosal epithelial progenitor cells (EPCs) in the rat vagina. Rat vagina was obtained from f77emale Sprague-Dawley rats (12 weeks old, n = 20). To identify EPCs in vagina, we followed a single-cell isolation protocol and analyzed the number of cells staining positive for EPC markers by flow cytometry. The expression of EPC-specific markers (CD44, ERα, PR) was evaluated by immunofluorescence. As shown by confocal immunofluorescence, CD44/ERα double-labeled cells were mainly expressed in the basal cell and suprabasal cell layers. Immunofluorescent staining of CD44 was expressed in the plasma membrane of the vaginal epithelium, and ERα was expressed in the cytoplasm of the vaginal epithelium. The CD44/ERα double-positive cells were noted in the rat vagina by flow cytometric analysis. PR-labeled cells were localized in the intermediate layer of the epithelial space of the vagina. The results revealed the existence of EPCs in the vagina. These findings imply that resident EPCs may have a role in the regeneration of vaginal mucosa.


Assuntos
Células Epiteliais/citologia , Células-Tronco/citologia , Vagina/citologia , Animais , Células Epiteliais/metabolismo , Epitélio/metabolismo , Receptor alfa de Estrogênio/metabolismo , Feminino , Receptores de Hialuronatos/metabolismo , Ratos , Ratos Sprague-Dawley , Células-Tronco/metabolismo , Vagina/metabolismo
19.
Int J Pharm ; 554: 276-283, 2019 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-30423417

RESUMO

The aim of this work is to test the in vivo behavior of a mucoadhesive vaginal emulsion resistant to the clearance of vaginal fluids using ciprofloxacin (CPX) as an anti-infective model of drug. CPX is a broad-spectrum antibiotic used in the treatment of sexual tissues infections, as intravenous injection in a dose of 20 mg every 12 h. In this study, CPX was incorporated in water in silicone (W/S) mucoadhesive emulsions and the in vivo residence time and the CPX in vivo absorption and distribution to the sexual organs was studied using the rat as animal model. W/S emulsion shows excellent in vitro bioadhesion having high resistance to the vaginal fluids clearance. The drug release profiles show a constant release of CPX during at least 6 h according to a zero-order kinetics. In vivo computerized PET/CT Image Analysis after intravaginal administration to rats indicates that W/S emulsions remain in the vaginal area for a long time and shows a good absorption of the radiotracers used as markers through the vaginal mucosa. Ciprofloxacin pharmacokinetic studies developed after the single intravaginal administration of W/S emulsion shows a good absorption and distribution of CPX on the uterus and ovarian tissue. A significant concentration of CPX in the sexual tissues was observed after 24 h of administration of W/S emulsion. Therefore, W/S emulsions have a good in vivo residence and drug release in the vaginal mucosae showing a great potential for the treatment of sexual tissues infections, as vaginal bioadhesive delivery systems of antinfectious drugs.


Assuntos
Antibacterianos/administração & dosagem , Ciprofloxacino/administração & dosagem , Silicones/química , Vagina/metabolismo , Adesividade , Administração Intravaginal , Animais , Antibacterianos/farmacocinética , Química Farmacêutica/métodos , Ciprofloxacino/farmacocinética , Preparações de Ação Retardada , Liberação Controlada de Fármacos , Emulsões , Feminino , Membrana Mucosa/metabolismo , Tomografia Computadorizada com Tomografia por Emissão de Pósitrons , Ratos , Ratos Endogâmicos WKY , Distribuição Tecidual , Água/química
20.
Int Urogynecol J ; 30(3): 439-446, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29600404

RESUMO

INTRODUCTION AND HYPOTHESIS: Pelvic organ prolapse (POP) is a multifactorial disorder that impairs the quality of life (QoL) of older women in particular. The purpose of this study was to elucidate the pathogenesis of POP by focusing on the extracellular matrix (ECM). METHODS: Patients were classified into two groups-with or without cervical elongation-using the POP quantification system. Specimens were obtained from 29 women with POP during hysterectomy. The expression of fibulin-5, elastin, integrin ß1 (ITGß1), lysyl oxidase-like protein-1 (LOXL1) and collagen in the vagina, uterosacral ligament, and uterine cervix was investigated by quantitative real-time polymerase chain reaction (RT-PCR) and correlation between gene levels and severity of POP examined. The location of proteins was analyzed using immunohistochemical staining and expression of fibulin-5 protein analyzed by Western blotting. RESULTS: Fibulin-5 and elastin were mainly expressed in lamina propria and fibromuscular layers of the vagina and uterosacral ligament. Gene levels of fibulin-5 and ITGß1 in uterosacral ligaments increased with severity of POP in women with cervical elongation, while no correlation was observed in women with a normal cervix. In women with uterine cervical elongation, each ECM-related gene significantly increased with POP staging. Furthermore, fibulin-5 protein also increased in the uterosacral ligament and uterine cervix. CONCLUSIONS: The severity of POP and gene expression of ECM-related proteins were inversely correlated in vaginal tissue in a normal and elongated cervix. These results suggested that the differing progression of the two types of POP have a relationship with ECM-related protein.


Assuntos
Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Ligamentos/metabolismo , Prolapso de Órgão Pélvico/genética , Prolapso de Órgão Pélvico/metabolismo , Vagina/metabolismo , Idoso , Aminoácido Oxirredutases/genética , Aminoácido Oxirredutases/metabolismo , Colo do Útero/metabolismo , Colo do Útero/patologia , Colágeno/genética , Colágeno/metabolismo , Elastina/genética , Elastina/metabolismo , Feminino , Humanos , Integrina beta1/genética , Integrina beta1/metabolismo , Pessoa de Meia-Idade , Prolapso de Órgão Pélvico/patologia , RNA/metabolismo , Índice de Gravidade de Doença , Transcriptoma
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