RESUMO
Cryptococcus gattii is the causative agent of cryptococcosis infection that can lead to pneumonia and meningitis in immunocompetent individuals. The molecular basis of the pathogenic process and impact on the host biochemistry are poorly understood and remain largely unknown. In this context, a comparative proteomic analysis was performed to investigate the response of the host during an infection caused by C. gattii. Lungs of experimentally infected rats were analyzed by shotgun proteomics to identify differentially expressed proteins induced by C. gattii clinical strain. The proteomic results were characterized using bioinformatic tools, and subsequently, the molecular findings were validated in cell culture and lungs of infected animals. A dramatic change was observed in protein expression triggered by C. gattii infection, especially related to energy metabolism. The main pathways affected include aerobic glycolysis cycle, TCA cycle, and pyrimidine and purine metabolism. Analyses in human lung fibroblast cells confirmed the altered metabolic status found in infected lungs. Thus, it is clear that C. gattii infection triggers important changes in energy metabolism leading to the activation of glycolysis and lactate accumulation in lung cells, culminating in a cancerlike metabolic status known as the Warburg effect. The results presented here provide important insights to better understand C. gattii molecular pathogenesis.
Assuntos
Criptococose/metabolismo , Metabolismo Energético/fisiologia , Glicólise/fisiologia , Pulmão/metabolismo , Proteoma/metabolismo , Proteômica/métodos , Animais , Linhagem Celular , Criptococose/microbiologia , Cryptococcus gattii/fisiologia , Modelos Animais de Doenças , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/microbiologia , Interações Hospedeiro-Patógeno , Humanos , Pulmão/microbiologia , Masculino , Ratos WistarRESUMO
The pivotal role of human endometrial stromal cells (hESCs) in the development of endometriosis lies in their ability to adopt a pro-invasive and proinflammatory profile upon migration to areas outside the uterus. However, the molecular mechanisms involved in these events remain unclear. In this study, we investigated how angiotensin II (Ang II) affects the plasminogen-plasmin system in hESCs, and the mechanisms underlying cell proliferation, migration, matrix degradation, and inflammation. Precursors, receptors, and peptidases involved in angiotensin metabolism increased significantly in Ang II-treated hESCs. The expression and activity of tissue (tPA)- and urokinase (uPA)- type plasminogen activators and the receptor for uPA (uPAR) were induced in the presence of Ang II. The up-regulation of tPA-uPA/uPAR pathway significantly contributes to heightened plasmin production both on the surface of hESCs and in their conditioned media. As a result, the plasmin generation induced by Ang II enhances the degradation of fibrin and matrix proteins, while also boosting hESC viability, proliferation, and migration through the up-regulation of growth factor expression. Notably, Ang II-induced hESC migration was dependent on the generation of active plasmin on cell surface. Ang II regulates oxidative and inflammatory signalling in hESCs primarily via NADPH oxidase and through the up-regulation of proinflammatory cytokines and adhesion molecules. Interestingly, Ang II receptor (AT1R) blockage, decreased plasmin generation, tPA-uPA/uPAR expression and hESC migration. Our results suggest that Ang II/AT1R axis regulates hESC proliferation and migration through tPA-uPA/uPAR pathway activation and plasmin generation. We propose the Ang II/AT1R axis as a potential target for endometriosis treatment.
Assuntos
Angiotensina II , Movimento Celular , Endométrio , Matriz Extracelular , Fibrinolisina , Plasminogênio , Receptor Tipo 1 de Angiotensina , Transdução de Sinais , Células Estromais , Humanos , Feminino , Endométrio/metabolismo , Endométrio/citologia , Endométrio/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Movimento Celular/fisiologia , Fibrinolisina/metabolismo , Células Estromais/metabolismo , Células Estromais/efeitos dos fármacos , Angiotensina II/farmacologia , Transdução de Sinais/fisiologia , Transdução de Sinais/efeitos dos fármacos , Matriz Extracelular/metabolismo , Matriz Extracelular/efeitos dos fármacos , Receptor Tipo 1 de Angiotensina/metabolismo , Plasminogênio/metabolismo , Células Cultivadas , Inflamação/metabolismoRESUMO
The pathophysiology of recurrent pregnancy loss (RPL) involves deficiencies in the proliferation and migration capacities of endometrial stromal cells (hESCs), which impair embryo implantation and development. Since animal venoms are rich source of bioactive molecules, we aimed to characterize the cytoprotective effects of Lonomia obliqua venom on hESCs. hESCs were isolated from endometrial biopsies and the mechanisms of L. obliqua venomous secretions on cell viability, proliferation and migration were characterized. Venom components were identified by chromatography and proteomic analyses. L. obliqua venom induced hESC proliferation, viability and migration in a dose-dependent manner, both in the presence and absence of serum. By ion-exchange chromatography, one fraction enriched in cytoprotective components and devoid of hemotoxins was obtained. Venom proteome identified at least six protein classes with potential cytoprotective properties (hemolins, lipocalins, hemocyannins, antiviral proteins, antimicrobial peptides, and protease inhibitors). L. obliqua venom protected hESCs from oxidative insult. Cytoprotection was also related to nitric oxide and PKC-ERK-activation and down-regulation of cAMP-PKA-dependent pathways that control cell proliferation. L. obliqua venom-induced hESC viability, proliferation and migration occurs mainly by protecting against oxidative damage and activating ERK. Thus, L. obliqua venom components are promising pharmacological tools to understand the underlying mechanisms of hESC deficiency in RPL.
Assuntos
Venenos de Artrópodes , Animais , Humanos , Venenos de Artrópodes/química , Proteômica , Células EpiteliaisRESUMO
AIM: In this study, we investigated how platelets and aorta contribute to the creation and maintenance of a prothrombotic state in an experimental model of postmenopausal hypertension in ovariectomized rats. METHODS: Bilateral ovariectomy was performed in both 14-week-old female spontaneously hypertensive (SHR) and normotensive Wistar Kyoto (WKY) rats. The animals were kept in phytoestrogen free diet. Vascular parameters, platelet, coagulation and aortic prothrombotic functions and mechanisms were assessed. RESULTS: Exacerbated platelet aggregation was observed in both SHR and WKY animals after ovariectomy. The mechanism was related to aortic COX2 downregulation and reduction in AMP, ADP, and ATP hydrolysis in serum and platelets. A procoagulant potential was observed in plasma from ovariectomized rats and this was confirmed by kallikrein and factor Xa generation in aortic rings. Aortic rings derived from ovariectomized SHR presented a greater thrombin generation capacity compared to equivalent rings from WKY animals. The mechanism involved tissue factor and PAR-1 upregulation as well as an increase in extrinsic coagulation and fibrinolysis markers in aorta and platelets. Aortic smooth muscle cells pre-treated with a plasma pool derived from estrogen-depleted animals developed a procoagulant profile with tissue factor upregulation. This procoagulant profile was dependent on inflammatory signalling, since NFκB inhibition attenuated the procoagulant activity and tissue factor expression. CONCLUSIONS: A prothrombotic phenotype was observed in both WKY and SHR ovariectomized rats being associated with platelet hyperreactivity and tissue factor upregulation in aorta and platelets. The mechanism involves proinflammatory signalling that supports greater thrombin generation in aorta and vascular smooth muscle cells.
Assuntos
Hipertensão , Trombina , Ratos , Feminino , Animais , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Trombina/metabolismo , Trombina/farmacologia , Tromboplastina , Hipertensão/metabolismo , Aorta , EstrogêniosRESUMO
The impact of Chlamydia trachomatis (CT) infection on female's fertility is not completely established yet, since the level of evidence associating these factors is still weak. Hence, the goal of the present review is to contribute to a better elucidation of this matter. The electronic database chosen was the Medline/PubMed, with the last survey on May 11, 2021. Publication date was used as a filter, with the previous 5 years having been selected. The following describers were used: chlamydia trachomatis AND infertility; chlamydia trachomatis AND tubal alteration AND infertility; chlamydia AND low pregnancy rates. From the 322 studies screened, 293 that failed to meet our eligibility criteria were excluded. Subsequently, we removed seven studies for not having the possible correlation between CT infections and female infertility as its main focus, and three for being about sexually transmitted infections (STIs) in general. Moreover, two studies designed as reviews were also excluded. Ergo, we included 17 studies in our qualitative analysis. The authors conducted research individually and analyzed carefully the studies selected. As we retrieved the information needed for our study through reading the texts, no contact was made with the authors of the studies selected. This systematic review corroborates the hypothesis that CT infection potentiates female infertility, as 76.47% of the included studies found a positive correlation between them. We conclude that there is an important association between CT infection and female infertility. Ergo, making CT screening part of the infertility investigation routine is relevant and has a reasonable justification.
O impacto da infecção por Chlamydia trachomatis (CT) na fertilidade feminina ainda não está completamente estabelecido, uma vez que o nível de evidência associando esses fatores ainda é insignificante. Assim, o objetivo desta revisão é contribuir para uma melhor elucidação deste assunto. A base de dados eletrônica escolhida foi a Medline/PubMed, com a última pesquisa em 11 de maio de 2021. Utilizou-se como filtro a data de publicação, sendo selecionados os 5 anos anteriores. Foram usados os seguintes descritores: Chlamydia trachomatis E infertility; Chlamydia trachomatis E tubal alteration E infertility; Chlamydia E low pregnancy rates. Dos 322 estudos selecionados, 293 que não atenderam aos nossos critérios de elegibilidade foram excluídos. Posteriormente, retiramos sete estudos por não terem como foco principal a possível correlação entre infecção por CT e infertilidade feminina e três por tratarem de infecções sexualmente transmissíveis (ISTs) em geral. Além disso, dois estudos concebidos como revisões também foram excluídos. Portanto, incluímos 17 estudos em nossa análise qualitativa. Os autores realizaram pesquisas individualmente e analisaram criteriosamente os estudos selecionados. Como obtivemos as informações necessárias para nosso estudo por meio da leitura dos textos, nenhum contato foi feito com os autores. Esta revisão sistemática corrobora a hipótese de que a infecção por CT potencializa a infertilidade feminina, pois 76,47% dos estudos incluídos encontraram correlação positiva entre eles. Concluímos que existe uma associação importante entre infecção por CT e infertilidade feminina. Portanto, tornar os procedimentos de triagem por CT parte da rotina de investigação de infertilidade é relevante e justificável.
Assuntos
Infecções por Chlamydia , Infertilidade Feminina , Infecções por Chlamydia/complicações , Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis , Feminino , Fertilidade , Humanos , Infertilidade Feminina/complicações , Programas de Rastreamento , GravidezRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Acmella oleracea (L.) R. K. Jansen (Asteraceae), known as jambú in Brazil, is used in traditional medicine as analgesic and for inflammatory conditions, characterized by the presence of N-alkylamides, mainly spilanthol. This bioactive compound is responsible for the above-described pharmacological properties, including sialagogue and anesthetic. AIM OF THE STUDY: This study aimed to characterize the anti-inflammatory effects of A. oleracea leaves (AOEE-L) and flowers (AOEE-F) extracts, including an isolated alkylamide (spilanthol), using in vitro and in vivo models. The mechanism underlying this effect was also investigated. MATERIALS AND METHODS: Extracts were analyzed by HPLC-ESI-MS/MS in order to characterize the N-alkylamides content. AOEE-L, AOEE-F (25-100 µg/mL) and spilanthol (50-200 µM) were tested in vitro on VSMC after stimulation with hyperglycemic medium (25 mM glucose). Their effects over nitric oxide (NO) generation, chymase inhibition and expression, catalase (CAT), superoxide anion (SOD) radical activity were evaluated. After an acute administration of extracts (10-100 mg/mL) and spilanthol (6.2 mg/mL), the anti-inflammatory effects were evaluated by applying the formalin test in rats. Blood was collected to measure serum aminotransferases activities, NO activity, creatinine and urea. RESULTS: A number of distinct N-alkylamides were detected and quantified in AOEE-L and AOEE-F. Spilanthol was identified in both extracts and selected for experimental tests. Hyperglycemic stimulation in VSMC promoted the expression of inflammatory parameters, including chymase, NO, CAT and SOD activity and chymase expression, all of them attenuated by the presence of the extracts and spilanthol. The administration of extracts or spilanthol significantly inhibited edema formation, NO production and cell tissue infiltration in the formalin test, without causing kidney and liver toxicity. CONCLUSION: Taken together, these results provide evidence for the anti-inflammatory activity of leaves and flowers extracts of jambú associated distinctly with their chemical profile. The effects appear to be associated with the inhibition of chymase activity, suppression of the proinflammatory cytokine NO and antioxidant activities.
Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Asteraceae/química , Quimases/antagonistas & inibidores , Extratos Vegetais/farmacologia , Alcamidas Poli-Insaturadas/farmacologia , Animais , Anti-Inflamatórios/uso terapêutico , Antioxidantes/química , Antioxidantes/uso terapêutico , Brasil , Linhagem Celular , Quimases/metabolismo , Edema/induzido quimicamente , Edema/tratamento farmacológico , Edema/patologia , Etanol/química , Flores/química , Formaldeído/toxicidade , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/patologia , Masculino , Medicina Tradicional , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/metabolismo , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Folhas de Planta/química , Alcamidas Poli-Insaturadas/uso terapêutico , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismoRESUMO
INTRODUCTION: Despite recent advances in assisted reproduction techniques and recent knowledge regarding embryo and endometrium quality, implantation and birth rates remain low. The objective of this study was to investigate whether clomiphene citrate alters endometrial maturation in infertile patients. METHODS: In a prospective self-matched cohort study, we assessed the ovulation of women in spontaneous and stimulated cycles (with clomiphene citrate). We determined the ovulation day by ultrasound scanning. In both cycles, we took four blood samples (BS1 - at early proliferative phase, BS2 - at mid proliferative phase, BS3 - after ovulation and BS4 - at mid luteal phase) to determine the serum concentrations of FSH, LH, estradiol and progesterone. We retrieved an endometrial biopsy five days after ovulation, followed by blinded analysis and classification according to Noyes criteria, in both cycles. RESULTS: Twenty-two participants completed the study. There were significant differences in FSH BS3 (p=0.001), in LH BS3 and BS4 (p<0.001 and p=0.049, respectively), in estradiol BS2, BS3 and BS4 (p<0.001, p=0.024 and p<0.001, respectively) and in progesterone BS3 and BS4 (p=0.028 and p<0.001, respectively). Considering Noyes criteria, there was a one-day delay when comparing the stimulated cycle with the spontaneous cycle (p=0.004), and a two-day delay when comparing the stimulated cycle with the biopsy day. CONCLUSION: This study indicates that ovarian stimulation with clomiphene citrate delays the endometrial maturity, and could possibly impair the implantation process due to asynchrony.
Assuntos
Clomifeno , Indução da Ovulação , Estudos de Coortes , Endométrio/diagnóstico por imagem , Estradiol , Feminino , Humanos , Progesterona , Estudos ProspectivosRESUMO
AIMS: Accidental contact with the Lonomia obliqua caterpillar is a common event in southern Brazil. Envenomed victims present consumption coagulopathy, which can evolve to acute kidney injury (AKI). In the present study, we searched for AKI biomarkers and changes in molecular pathway signatures through urine proteomic analysis. METHODOLOGY: Male Wistar rats were injected with L. obliqua venom (1.5 mg/kg, via s.c.) or 0.9 % NaCl and distributed into metabolic cages. After 24 h, urine was obtained, and the set of differentially regulated proteins was analyzed by MudPIT technology in an OrbiTRAP mass spectrometer. RESULTS: L. obliqua venom leads to an increase in urine output and water and electrolyte excretion and to an increase in the albumin to creatine ratio in urine. The proteomic analysis revealed an up-regulation of tubular injury biomarkers, such as neutrophil-gelatinase associated lipocalin (NGAL) and cystatin C, in urine from envenomed rats. Several components related to the heme scavenging system were up-regulated or exclusively identified in urine from envenomed animals. There was an increase in urinary heme levels and hemoglobin subunits, hemopexin, haptoglobin, and biliverdin reductase. Similarly, kinin- and angiotensin-generating/degrading peptidases, such as kallikreins, neprilysin, plasmin, dipeptidyl peptidase IV, cathepsin D, kininogen, and neutral, basic, glutamyl, and acidic aminopeptidases, were also up-regulated in urine. CONCLUSIONS: L. obliqua envenomation induced tubular and glomerular injury, probably involving heme/hemoglobin toxicity and an imbalance in the kinin/angiotensin generating/degrading system.
Assuntos
Injúria Renal Aguda/induzido quimicamente , Aminopeptidases/metabolismo , Venenos de Artrópodes/toxicidade , Hemoglobinúria , Lepidópteros , Proteômica , Aminopeptidases/química , Animais , Heme , Hemoglobinas , Larva/fisiologia , Masculino , Ratos , Ratos Wistar , Urinálise , Urina/químicaRESUMO
OBJECTIVE: The aim of this study was to investigate the efficacy of protocols for mice ovary cryopreservation to compare the differences in Mouse Vasa Homologue expression (a germline cell marker) and ovarian viability after vitrification or slow freezing. METHODS: Female CF1 mice aged 40-45 days were randomly divided into three groups: Control, vitrification or slow freezing. Their ovaries were surgically removed, rinsed in saline solution and cryopreserved. For vitrification, we used a commercial protocol and for slow freeze, we used 1.5 M ethylene glycol (EG) as cryoprotectant. After that, the ovaries were processed for histological an immunohistochemical analysis, and counting of primordial, primary, pre-antral and antral follicles. RESULTS: No significant difference was found in the proportion of high-quality primordial, primary and pre-antral follicles after thawing/warming in the slow freezing and vitrification groups. The immunohistochemistry for MVH antibody demonstrated that the slow freeze group had a higher number of unmarked cells (p=0.012), indicating a harmful effect on the MVH expression in the ovarian tissue, where the cell structure is complex. CONCLUSION: Although both protocols indicated similar results in the histological analysis of follicular counts, the vitrification protocol was significantly better to preserve ovarian stem cells, an immature germ cell population. These cells are able to self-renew having regeneration potential, and may be effective for the treatment of ovarian failure and consequently infertility.
Assuntos
Criopreservação/métodos , Preservação da Fertilidade/métodos , Ovário , Vitrificação , Animais , Feminino , Camundongos , Ovário/citologia , Ovário/fisiologia , Células-Tronco/citologia , Células-Tronco/fisiologiaRESUMO
Kallikrein-kinin system (KKS) is involved in vascular reactivity and inflammatory response to cytotoxic drugs. Since cisplatin is a widely used chemotherapy and its cytotoxic mechanism can trigger inflammation and oxidative damage, in this work we evaluated the role of KKS in an animal model of cisplatin-induced ovarian toxicity. Biomarkers of ovarian stem cells, activity of KKS, inflammation and oxidative damage were measured in ovarian tissue of C57BL/6 female mice treated with vehicle or cisplatin (2.5â¯mg/kg). Cisplatin group presented greater number of atretic follicles, and lower numbers of antral and total viable follicles. Ki67, DDX4 and OCT-4 markers were similar between groups. Cisplatin triggered plasma and ovarian tissue kallikrein generation; and increased expression of bradykinin receptors B1 and B2. Neutrophil and macrophage infiltration markers increased. Superoxide anion generation also increased, while reduced glutathione levels decreased. These results suggest that KKS is activated and contributes to ovarian injury during cisplatin treatment.
Assuntos
Antineoplásicos/efeitos adversos , Cisplatino/efeitos adversos , Ovário/efeitos dos fármacos , Animais , Feminino , Sistema Calicreína-Cinina , Calicreínas/metabolismo , Camundongos Endogâmicos C57BL , Óxido Nítrico/metabolismo , Ovário/metabolismo , Ovário/patologia , Estresse Oxidativo/efeitos dos fármacos , Receptor B1 da Bradicinina/metabolismo , Receptor B2 da Bradicinina/metabolismoRESUMO
Some biomaterial scaffolds can positively interfere with tissue regeneration and are being developed to successfully repair the tissue function. The possibility of using epithelial cells combined with biomaterials appears to be a new option as therapeutic application. This combination emerges as a possibility for patients with Mayer-Rokitansky-Kuster-Hauser syndrome which requires vaginal repair and can be performed with tissue-engineered solution containing cells and biomaterials. It is expected that tissue-engineered solution containing cells and biomaterials would promote tissue repair in a more efficient, modern, and safe way. This study tested the efficiency of tissue-engineered solution containing human malignant melanoma cell line (HMV-II) and different biomaterials, including Cellprene®, Membracel®, and poly lactic-co-glycolic acid/epoxidized polyisoprene. The cells adhered better on poly lactic-co-glycolic acid/epoxidized polyisoprene, and it was found that tissue-engineered solution may also contain mesenchymal stem cells cultivated on poly lactic-co-glycolic acid/epoxidized polyisoprene. Histological, immunofluorescence, and scanning electron microscopy analyses were performed. These initial in vitro results suggest that tissue-engineered solution containing cells and poly lactic-co-glycolic acid/epoxidized polyisoprene is a potential for tissue reconstruction.
Assuntos
Regeneração Tecidual Guiada/métodos , Procedimentos de Cirurgia Plástica/métodos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Engenharia Tecidual/métodos , Alicerces Teciduais , Transtornos 46, XX do Desenvolvimento Sexual/cirurgia , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Linhagem Celular , Anormalidades Congênitas/cirurgia , Células Epiteliais , Feminino , Humanos , Células-Tronco Mesenquimais , Ductos Paramesonéfricos/anormalidades , Ductos Paramesonéfricos/cirurgia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/farmacologia , SoluçõesRESUMO
BACKGROUND: Lonomia obliqua venom is nephrotoxic and acute kidney injury (AKI) is the main cause of death among envenomed victims. Mechanism underlying L. obliqua-induced AKI involves renal hypoperfusion, inflammation, tubular necrosis and loss of glomerular filtration and tubular reabsorption capacities. In the present study, we aimed to investigate the contribution of kallikrein to the hemodynamic instability, inflammation and consequent renal and vascular impairment. METHODOLOGY/PRINCIPAL FINDINGS: Addition of L. obliqua venom to purified prekallikrein and human plasma in vitro or to vascular smooth muscle cells (VSMC) in culture, was able to generate kallikrein in a dose-dependent manner. Injected in rats, the venom induced AKI and increased kallikrein levels in plasma and kidney. Kallikrein inhibition by aprotinin prevented glomerular injury and the decrease in glomerular filtration rate, restoring fluid and electrolyte homeostasis. The mechanism underlying these effects was associated to lowering renal inflammation, with decrease in pro-inflammatory cytokines and matrix metalloproteinase expression, reduced tubular degeneration, and protection against oxidative stress. Supporting the key role of kallikrein, we demonstrated that aprotinin inhibited effects directly associated with vascular injury, such as the generation of intracellular reactive oxygen species (ROS) and migration of VSMC induced by L. obliqua venom or by diluted plasma obtained from envenomed rats. In addition, kallikrein inhibition also ameliorated venom-induced blood incoagulability and decreased kidney tissue factor expression. CONCLUSIONS/SIGNIFICANCE: These data indicated that kallikrein and consequently kinin release have a key role in kidney injury and vascular remodeling. Thus, blocking kallikrein may be a therapeutic alternative to control the progression of venom-induced AKI and vascular disturbances.
Assuntos
Injúria Renal Aguda/induzido quimicamente , Venenos de Artrópodes/toxicidade , Calicreínas/antagonistas & inibidores , Mariposas/fisiologia , Injúria Renal Aguda/prevenção & controle , Animais , Aprotinina , Transtornos da Coagulação Sanguínea/induzido quimicamente , Modelos Animais de Doenças , Taxa de Filtração Glomerular , Larva/fisiologia , Masculino , Ratos , Ratos WistarRESUMO
The recent microcephaly outbreak in Brazil has been associated with Zika virus (ZIKV) infection. The current understanding of damage caused by ZIKV infection is still unclear, since it has been implicated in other neurodegenerative and developmental complications. Here, the differential proteome analysis of human mesenchymal stem cells (hMSC) infected with a Brazilian strain of ZIKV was identified by shotgun proteomics (MudPIT). Our results indicate that ZIKV induces a potential reprogramming of the metabolic machinery in nucleotide metabolism, changes in the energy production via glycolysis and other metabolic pathways, and potentially inhibits autophagy, neurogenesis, and immune response by downregulation of signaling pathways. In addition, proteins previously described in several brain pathologies, such as Alzheimer's disease, autism spectrum disorder, amyotrophic lateral sclerosis, and Parkinson's disease, were found with altered expression due to ZIKV infection in hMSC. This potential link between ZIKV and several neuropathologies beyond microcephaly is being described here for the first time and can be used to guide specific follow-up studies concerning these specific diseases and ZIKV infection.
Assuntos
Células-Tronco Mesenquimais/metabolismo , Doenças do Sistema Nervoso/patologia , Doenças do Sistema Nervoso/virologia , Infecção por Zika virus/metabolismo , Infecção por Zika virus/patologia , Zika virus/fisiologia , Adulto , Feminino , Humanos , Proteoma/metabolismoRESUMO
Mesenchymal stem cells (MSCs) have desirable characteristics for use in therapy in animal models and veterinary medicine, due to their capacity of inducing tissue regeneration and immunomodulation. The objective of this study was to evaluate the differences between canine adipose tissue-derived MSCs (AD-MSCs) extracted from subcutaneous (Sc) and visceral (Vs) sites. Surface antigenic markers, in vitro differentiation, and mineralized matrix quantification of AD-MSCs at different passages (P4, P6, and P8) were studied. Immunophenotypic analysis showed that AD-MSCs from both sites were CD44+, CD90+, and CD45-. Moreover, they were able, in vitro, to differentiate into fat, cartilage, and bone. Sc-AD-MSCs preserve in vitro multipotentiality up to P8, but Vs-AD-MSCs only tri-differentiated up to P4. In addition, compared to Vs-AD-MSCs, Sc-AD-MSCs had greater capacity for in vitro mineralized matrix synthesis. In conclusion, Sc-AD-MSCs have advantages over Vs-AD-MSCs, as Sc AD-MSCs preserve multipotentiality during a greater number of passages, have more osteogenic potential, and require less invasive extraction.
Assuntos
Diferenciação Celular , Gordura Intra-Abdominal/citologia , Células-Tronco Mesenquimais/citologia , Gordura Subcutânea/citologia , Animais , Cães , Feminino , Imunofenotipagem/veterináriaRESUMO
To investigate the effects of heterologous adipose-derived stem cells (ADSCs) on autologous full-thickness skin grafts, we designed a first-intention healing model using Wistar rats. We harvested and sutured two full-thickness skin grafts in the dorsal recipient beds of 15 rats, randomized into three groups. In the treatment group, 1â¯×â¯106 ADSCs resuspended in saline solution (200⯵L) were administered subcutaneously to the skin graft. The control group received only saline solution subcutaneously, whereas the negative control group did not receive any treatment. Compressive dressings were maintained until postoperative day 5. The grafts were assessed by two observers, who checked for the presence of epidermolysis on day 14. Planimetry showed the relative areas of normal skin, redness, ulceration, and contraction. Graft samples were obtained on day 14 and stained with hematoxylin and eosin and Masson's trichrome. Epidermal analysis evaluated thickening, keratosis, acanthosis, hydropic degeneration, and inflammatory infiltrate. Dermal evaluation investigated the absence of hair follicles, granulation tissue formation, presence of inflammatory infiltrate, and collagen deposition. Immunohistochemistry was performed for dermal anti-VEGF and epidermal anti-Ki-67 staining. The ADSC group presented better macroscopic aspects, lower incidence of epidermolysis, and less loss of hair follicles. In addition, the ADSC group presented the lowest frequency of histopathological changes in the dermis and epidermis, as well as the largest subcutaneous and granulation tissue VEGF averages and the weakest Ki-67 staining of the epidermal basal layer. Subcutaneous administration of ADSCs may improve the integration of skin grafts, reducing the deleterious effects of ischemia and reperfusion injury.
Assuntos
Adipócitos/citologia , Transplante de Pele , Transplante de Células-Tronco , Tecido Adiposo/citologia , Animais , Modelos Animais de Doenças , Ratos , Ratos Wistar , Pele , Células-Tronco , Engenharia Tecidual , CicatrizaçãoRESUMO
Abstract The impact of Chlamydia trachomatis (CT) infection on female's fertility is not completely established yet, since the level of evidence associating these factors is still weak. Hence, the goal of the present review is to contribute to a better elucidation of this matter. The electronic database chosen was the Medline/PubMed, with the last survey on May 11, 2021. Publication date was used as a filter, with the previous 5 years having been selected. The following describers were used: chlamydia trachomatis AND infertility; chlamydia trachomatis AND tubal alteration AND infertility; chlamydia AND low pregnancy rates. From the 322 studies screened, 293 that failed to meet our eligibility criteria were excluded. Subsequently, we removed seven studies for not having the possible correlation between CT infections and female infertility as its main focus, and three for being about sexually transmitted infections (STIs) in general. Moreover, two studies designed as reviews were also excluded. Ergo, we included 17 studies in our qualitative analysis. The authors conducted research individually and analyzed carefully the studies selected. As we retrieved the information needed for our study through reading the texts, no contact was made with the authors of the studies selected. This systematic review corroborates the hypothesis that CT infection potentiates female infertility, as 76.47% of the included studies found a positive correlation between them. We conclude that there is an important association between CT infection and female infertility. Ergo, making CT screening part of the infertility investigation routine is relevant and has a reasonable justification.
Resumo O impacto da infecção por Chlamydia trachomatis (CT) na fertilidade feminina ainda não está completamente estabelecido, uma vez que o nível de evidência associando esses fatores ainda é insignificante. Assim, o objetivo desta revisão é contribuir para uma melhor elucidação deste assunto. A base de dados eletrônica escolhida foi a Medline/PubMed, com a última pesquisa em 11 de maio de 2021. Utilizou-se como filtro a data de publicação, sendo selecionados os 5 anos anteriores. Foram usados os seguintes descritores: Chlamydia trachomatis E infertility; Chlamydia trachomatis E tubal alteration E infertility; Chlamydia E low pregnancy rates. Dos 322 estudos selecionados, 293 que não atenderam aos nossos critérios de elegibilidade foram excluídos. Posteriormente, retiramos sete estudos por não terem como foco principal a possível correlação entre infecção por CT e infertilidade feminina e três por tratarem de infecções sexualmente transmissíveis (ISTs) em geral. Além disso, dois estudos concebidos como revisões também foram excluídos. Portanto, incluímos 17 estudos em nossa análise qualitativa. Os autores realizaram pesquisas individualmente e analisaram criteriosamente os estudos selecionados. Como obtivemos as informações necessárias para nosso estudo por meio da leitura dos textos, nenhum contato foi feito com os autores. Esta revisão sistemática corrobora a hipótese de que a infecção por CT potencializa a infertilidade feminina, pois 76,47% dos estudos incluídos encontraram correlação positiva entre eles. Concluímos que existe uma associação importante entre infecção por CT e infertilidade feminina. Portanto, tornar os procedimentos de triagem por CT parte da rotina de investigação de infertilidade é relevante e justificável.
Assuntos
Humanos , Feminino , Gravidez Tubária , Infecções Sexualmente Transmissíveis , Chlamydia trachomatisRESUMO
INTRODUCTION: The rationale of mesenchymal stem cells (MSCs) as a novel therapeutic approach in certain neurodegenerative diseases is based on their ability to promote neurogenesis. Hippocampal atrophy has been related to bipolar disorder (BD) in preclinical, imaging and postmortem studies. Therefore, the development of new strategies to stimulate the neurogenesis process in BD is crucial. OBJECTIVES: To investigate the behavioral and neurochemical changes induced by transplantation of MSCs in a model of mania-like behavior induced by lisdexamfetamine dimesylate (LDX). METHODS: Wistar rats (n=65) received one oral daily dose of LDX (10 mg/kg) or saline for 14 days. On the 8th day of treatment, the animals additionally received intrahippocampal saline or MSC (1 µL containing 25,000 cells) or lithium (47.5 mg/kg) as an internal experimental control. Two hours after the last administration, behavioral and neurochemical analyses were performed. RESULTS: LDX-treated rats had increased locomotor activity compared to saline-saline rats (p=0.004), and lithium reversed LDX-related hyperactive behavior (p<0.001). In contrast, the administration of MSCs did not change hyperlocomotion, indicating no effects of this treatment on LDX-treated rats (p=0.979). We did not find differences between groups in BDNF levels (p>0.05) in the hippocampus of rats. CONCLUSION: Even though these results suggest that a single intrahippocampal injection of MSCs was not helpful to treat hyperactivity induced by LDX and neither influenced BDNF secretion, we cannot rule out the possible therapeutic effects of MSCs. Further research is required to determine direct effects of LDX on brain structures as well as in other pathophysiological targets related to BD.
Assuntos
Transtorno Bipolar/terapia , Transplante de Células-Tronco Mesenquimais , Tecido Adiposo/citologia , Animais , Antimaníacos/farmacologia , Transtorno Bipolar/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Hipocampo/metabolismo , Hipocampo/cirurgia , Dimesilato de Lisdexanfetamina , Compostos de Lítio/farmacologia , Masculino , Camundongos Endogâmicos C57BL , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Estudo de Prova de Conceito , Ratos WistarRESUMO
Tissue engineering replaces injured tissues by manipulating cells, making scaffolds, and using molecules that stimulate the tissue. Mesenchymal stem cells (MSCs) are good candidates for tissue engineering, as this is one of the cell types which are recruited to repair injured tissues. Scaffolds are structural devices that allow cell fixation and migration, with polypropylene meshes being an example. This study aims to evaluate the culture of adipose tissue-derived mesenchymal stem cells (ADSCs), isolated from C57Bl/6 GFP + mice, in two types of polypropylene meshes (macroporous and microporous) in conventional culture plates and plates coated with methacrylate, over a period of fifteen days. The objective was to obtain the best interaction protocol between the mesh and the cells. The choice of the best method was based on adherence, maintenance of adherence and viability during culture. The amount of ADSCs adhering was checked daily by counting in a Neubauer Chamber and by using a growth curve performed with the MTT assay. The ADSCs adhering to the meshes were visualized with DAPI, panotic, hematoxylin and eosin, immunohistochemistry (integrin), and immunofluorescence (actin). ADSCs adhere to all forms of culture and to the two types of polypropylene mesh. ADSCs adhered more to the microporous mesh, within the seven day period of culture and in the plates without methacrylate. Thus, polypropylene meshes offer a good scaffold for ADSCs to adhere to.
A engenharia de tecidos substitui tecidos danificados com a manipulação de células, confecção de arcabouços e a utilização de moléculas que estimulem o tecido. As células-tronco mesenquimais (MSCs) são boas candidatas para engenharia de tecido, pois são um dos tipos celulares recrutadas para a reparação de tecidos lesionados. O arcabouço deve ser um dispositivo estrutural que forneça uma estrutura para o crescimento e a diferenciação celular no sítio, sendo a tela de polipropileno um exemplo. O objetivo deste estudo foi avaliar o cultivo de células-tronco mesenquimais de tecido de adiposo (ADSCs), isoladas de camundongos C57Bl/6 GFP+, em dois tipos de telas de polipropileno (macroporosa e microporosa) em placas de cultura convencionais e revestidas com metacrilato, durante quinze dias, para obter o melhor protocolo de interação entre a tela e as células. A escolha do melhor método foi baseada na adesão, manutenção da adesão e viabilidade durante cultivo. A quantidade de ADSCs aderidas foi verificada diariamente em contagem em Câmara de Neubauer e através de uma curva de crescimento realizada através de ensaio de MTT. As ADSCs aderidas nas telas foram visualizadas com a marcação de DAPI, panótico, hematoxilina e eosina, imumo-histoquímica (integrina) e imunofluorescência (actina). Nas duas formas de cultivo e nos dois tipos de telas de polipropileno houve aderência das ADSCs. Houve maior aderência na tela microporosa, no período de sete dias de cultivo e em placas sem metacrilato. Conclui-se que a tela de polipropileno oferece um bom arcabouço para as ADSCs se aderirem.
Assuntos
Animais , Camundongos , Polipropilenos/análise , Inclusão do Tecido/métodos , Engenharia Tecidual/métodos , Alicerces Teciduais , Células-Tronco Mesenquimais , CamundongosRESUMO
A engenharia de tecidos tem como objetivo substituir tecidos danificados, manipulando células, confecção de arcabouços e a utilização de moléculas que estimulem o tecido. A proposta deste estudo foi avaliar duas técnicas de cultivo de células-tronco mesenquimais (MSC) em diferentes placas de cultura, utilizando dois tipos de telas de polipropileno (macroporoso e microporoso), para obter as melhores condições de interação entre a tela e as células, e definir uma proposta de protético para engenharia de tecidos. As telas de polipropileno foram cultivadas com células-tronco mesenquimais de tecido adiposo (ADSCs) isoladas de camundongos C57B1/6 GFP+ durante quinze dias em placas revestidas com metacrilato ou não revestidas com metacrilato. A quantidade de ADSCs aderidas foram verificadas diariamente em Câmara de Neubauer e através de uma curva de crescimento realizada pelo ensaio MTT. As ADSCs aderidas às malhas foram visualizadas com marcação de DAPI, panóticas, hematoxilina e eosina imuno-histoquímica e imunofluorescência. O melhor protocolo foi na tela microporosa, no o período de sete dias de cultivo e em placas sem metacrilato. Conclui-se que a tela de polipropileno fornece um bom suporte para as ADSCs se aderirem podendo ser utilizada na engenharia de tecidos.
Tissue engineering replaces injured tissues by manipulating cells, making scaffolds, and using molecules that stimulate the tissue. Mesenchymal stem cells (MSCs) are good candidates for tissue engineering, as this is one of the cell types which are recruited to repair injured tissues. Scaffolds are structural devices that allow cell fixation and migration, with polypropylene meshes being an example. This study aims to evaluate the culture of adipose tissue-derived mesenchymal stem cells (ADSCs), isolated from C57Bl/6 GFP + mice, in two types of polypropylene meshes (macroporous and microporous) in conventional culture plates and plates coated with methacrylate, over a period of fifteen days. The objective was to obtain the best interaction protocol between the mesh and the cells. The choice of the best method was based on adherence, maintenance of adherence and viability during culture. The amount of ADSCs adhering was checked daily by counting in a Neubauer Chamber and by using a growth curve performed with the MTT assay. The ADSCs adhering to the meshes were visualized with DAPI, panotic, hematoxylin and eosin, immunohistochemistry (integrin), and immunofluorescence (actin). ADSCs adhere to all forms of culture and to the two types of polypropylene mesh. ADSCs adhered more to the microporous mesh, within the seven day period of culture and in the plates without methacrylate. Thus, polypropylene meshes offer a good scaffold for ADSCs to adhere to.