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1.
Article in English | WPRIM | ID: wpr-922760

ABSTRACT

Danshen-Chuanxiongqin Injection (DCI) is a commonly used traditional Chinese medicine for the treatment of cerebral ischemic stroke in China. However, its underlying mechanisms remain completely understood. The current study was designed to explore the protective mechanisms of DCI against cerebral ischemic stroke through integrating whole-transcriptome sequencing coupled with network pharmacology analysis. First, using a mouse model of cerebral ischemic stroke by transient middle cerebral artery occlusion (tMCAO), we found that DCI (4.10 mL·kg


Subject(s)
Brain Ischemia/genetics , Drugs, Chinese Herbal , Humans , Infarction, Middle Cerebral Artery/genetics , Ischemic Stroke , Myeloid Differentiation Factor 88/genetics , NF-kappa B/metabolism , Stroke/genetics , Toll-Like Receptor 2 , Toll-Like Receptor 4/metabolism
2.
Acta bioquím. clín. latinoam ; 54(3): 309-320, set. 2020. ilus, graf, tab
Article in Spanish | LILACS | ID: biblio-1130605

ABSTRACT

En los últimos años se ha determinado que los neutrófilos son células altamente versátiles y sofisticadas, cuyas funciones van mucho más allá de la eliminación de los microorganismos. En la infección con el Virus de Inmunodeficiencia Humana (HIV), si bien el papel de los neutrófilos no está totalmente caracterizado, actualmente está claro que la relación entre los neutrófilos y el virus es mucho más compleja de lo que se pensaba. Los objetivos de este trabajo fueron evaluar en pacientes con infección asintomática, y sin tratamiento antirretroviral, el efecto de la infección por el HIV sobre la muerte celular de los neutrófilos y la expresión de receptores de superficie. En pacientes seropositivos sin tratamiento hubo un aumento de la apoptosis temprana de los neutrófilos en relación a los grupos controles. Esta apoptosis aumentada no depende de la activación de la vía extrínseca o intrínseca. En estos pacientes hubo un aumento de la expresión de TLR2 que, unido al aumento de la apoptosis temprana, podría ser indicativo de un fenotipo activado de los neutrófilos. En conclusión, este trabajo aporta información sobre aspectos relacionados con la apoptosis de los neutrófilos en estadios tempranos de la infección por HIV, contribuyendo así a una mayor comprensión acerca del efecto de este virus sobre componentes de la respuesta inmune innata.


In recent years it has been determined that neutrophils are highly versatile and sophisticated cells whose functions go far beyond the elimination of microorganisms. In Human Immunodeficiency Virus (HIV) infection, the role of neutrophils is not fully characterized but it is now clear that the relationship between neutrophils and HIV is much more complex than previously thought. The aims of this study were to evaluate the effect of HIV infection on neutrophil cell death and the expression of surface molecules on neutrophils in patients with asymptomatic infection and without antiretroviral treatment (ART). In HIV seropositive patients without antiretroviral therapy there was an increase in the early apoptosis of neutrophils in relation to the control groups. This increased apoptosis does not depend on the activation of the extrinsic or intrinsic pathway. In these patients there was an increase in the expression of TLR2 which, together with the increase of early apoptosis, could be indicative of an activated phenotype of neutrophils. In conclusion, this study provides information on aspects related to the apoptosis of neutrophils in early stages of HIV infection and therefore contributes to a better understanding of the effect of this virus on components of the innate immune response.


Nos últimos anos, determinou-se que os neutrófilos são células altamente versáteis e sofisticadas, cujas funções vão muito além da eliminação dos microrganismos. Na infecção pelo HIV, embora o papel dos neutrófilos não esteja totalmente caracterizado, atualmente fica bem claro que a relação entre os neutrófilos e o vírus é muito mais complexa do que se pensava anteriormente. Os objetivos deste trabalho foram avaliar em pacientes com infecção assintomática, e sem tratamento antirretroviral, o efeito da infecção pelo HIV na morte celular dos neutrófilos e a expressão de receptores de superfície. Nos pacientes soropositivos sem tratamento, houve um aumento da apoptose precoce dos neutrófilos em relação aos grupos controle.Esta apoptose aumentada não depende da ativação da via extrínseca ou intrínseca. Nestes pacientes, houve um aumento da expressão de TLR2 que, juntamente com o aumento da apoptose precoce, poderia ser indicativo de um fenótipo ativado dos neutrófilos. Em conclusão, este trabalho fornece informações sobre aspectos relacionados com a apoptose dos neutrófilos em estágios precoces da infecção pelo HIV, contribuindo desse modo para uma maior compreensão sobre o efeito deste vírus nos componentes da resposta imune inata.


Subject(s)
Humans , Male , Female , Phenotype , Viruses , HIV Infections , HIV , Immunity, Innate , Neutrophils , Role , Therapeutics , HIV Antibodies/genetics , Cell Death , Apoptosis , Anti-Retroviral Agents , Toll-Like Receptor 2 , Asymptomatic Infections
3.
Arq. bras. med. vet. zootec. (Online) ; 72(4): 1329-1338, July-Aug. 2020. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1131492

ABSTRACT

The aim of this work was to measure HMGB1, TNF-alpha, and IL-8 in bronchoalveolar lavage (BAL), serum and TLR2 and TLR4mRNA expression in lung tissue of rabbits with two grades of acute lung injury (ALI). The animals were randomly assigned to groups with severe (S) and mild/moderate (MM) ALI, induced with warm saline, and a control group. HMGB1, TNF-alpha, IL-8, TLR2mRNA and TLR4mRNA were measured after ALI induction. The results showed increased levels of IL-8, TNF-alpha, HMGB1 and TLR4mRNA in the ALI groups. HMGB1, IL-8 and TNF-alpha concentrations in BAL were higher in S compared MM. Increased TLR4mRNA was observed in S and MM versus control. The results suggest an early participation of HMGB1 in ALI together with IL-8 and TNF-alpha and association with severity. TLR4 has early expression and role in ALI pathophysiology but is not associated with severity.(AU)


O objetivo deste trabalho é determinar os níveis de HMGB1, TNF-alfa e IL-8 no lavado broncoalveolar (BAL), bem como quantificar a expressão sérica de TLR2 e TLR4 mRNA em tecido pulmonar de coelhos com dois graus de lesão pulmonar aguda (LPA). Os animais foram distribuídos aleatoriamente em grupos com LPA grave (S) e leve / moderada (MM), induzidas com solução salina morna, e um grupo controle. HMGB1, TNF-alfa, IL-8, TLR2mRNA e TLR4mRNA foram medidos após a indução de LPA e quatro horas de ventilação mecânica. Os resultados mostraram níveis aumentados de IL-8, TNF-alfa, HMGB1 e TLR4mRNA nos grupos com LPA. As concentrações de HMGB1, IL-8 e TNF-alfa no LBA foram maiores no S comparado ao MM. Aumento de TLR4mRNA foi observado em S e MM versus controle. Os resultados sugerem uma participação precoce da HMGB1 na LPA em conjunto com IL-8 e TNF-alfa e associação com a gravidade da LPA. O TLR4 foi expresso na ALI e possivelmente possui papel precoce na fisiopatologia da LPA, mas sem associação com a gravidade.(AU)


Subject(s)
Animals , Rabbits , Cytokines , HMGB1 Protein , Acute Lung Injury , RNA, Messenger , Interleukin-8 , Tumor Necrosis Factor-alpha , Toll-Like Receptor 2 , Toll-Like Receptor 4
4.
Braz. oral res. (Online) ; 34: e012, 2020. graf
Article in English | LILACS | ID: biblio-1055530

ABSTRACT

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.


Subject(s)
Animals , Male , Periodontitis/etiology , Periodontitis/pathology , Toll-Like Receptor 2/antagonists & inhibitors , Lipopeptides/pharmacology , Osteoclasts/drug effects , Periodontitis/microbiology , Time Factors , Random Allocation , Alveolar Bone Loss/etiology , Alveolar Bone Loss/pathology , Disease Models, Animal , X-Ray Microtomography , Alveolar Process/drug effects , Alveolar Process/pathology , Tartrate-Resistant Acid Phosphatase , Gingiva/drug effects , Gingiva/pathology , Gingivitis/etiology , Gingivitis/pathology , Mice, Inbred C57BL
5.
Braz. oral res. (Online) ; 34: e012, 2020. graf
Article in English | LILACS | ID: biblio-1089395

ABSTRACT

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.


Subject(s)
Animals , Male , Periodontitis/etiology , Periodontitis/pathology , Toll-Like Receptor 2/antagonists & inhibitors , Lipopeptides/pharmacology , Osteoclasts/drug effects , Osteoclasts/physiology , Periodontitis/microbiology , Time Factors , Random Allocation , Alveolar Bone Loss/etiology , Alveolar Bone Loss/pathology , Disease Models, Animal , X-Ray Microtomography , Alveolar Process/drug effects , Alveolar Process/pathology , Tartrate-Resistant Acid Phosphatase , Gingiva/drug effects , Gingiva/pathology , Gingivitis/etiology , Gingivitis/pathology , Mice, Inbred C57BL
6.
Experimental Neurobiology ; : 547-553, 2019.
Article in English | WPRIM | ID: wpr-763791

ABSTRACT

Synucleinopathies are neurodegenerative disorders characterized by the progressive accumulation of α-synuclein (α-syn) in neurons and glia and include Parkinson's disease (PD) and dementia with Lewy bodies (DLB). In this review, we consolidate our key findings and recent studies concerning the role of Toll-like receptor 2 (TLR2), a pattern recognition innate immune receptor, in the pathogenesis of synucleinopathies. First, we address the pathological interaction of α-syn with microglial TLR2 and its neurotoxic inflammatory effects. Then, we show that neuronal TLR2 activation not only induces abnormal α-syn accumulation by impairing autophagy, but also modulates α-syn transmission. Finally, we demonstrate that administration of a TLR2 functional inhibitor improves the neuropathology and behavioral deficits of a synucleinopathy mouse model. Altogether, we present TLR2 modulation as a promising immunotherapy for synucleinopathies.


Subject(s)
Animals , Autophagy , Dementia , Immunotherapy , Lewy Bodies , Mice , Neurodegenerative Diseases , Neuroglia , Neurons , Neuropathology , Parkinson Disease , Toll-Like Receptor 2 , Toll-Like Receptors
7.
Braz. dent. j ; 29(1): 43-47, Jan.-Feb. 2018. tab, graf
Article in English | LILACS | ID: biblio-888722

ABSTRACT

Abstract The aim of this study was to evaluate the expression of MMP2 and MMP9 during apical periodontitis (AP) progression in TLR2 (TLR2 KO) and in MyD88 (MyD88 KO) knockout mice compared to wild type (WT) mice. AP was induced in mandibular first molars of TLR2 KO (n= 18), MyD88 KO (n= 18), and WT mice (n= 18). After 7, 21, and 42 days, the animals were euthanized and the jaws were dissected and subjected to histotechnical processing. Subsequent sections were stained by immunohistochemistry and evaluated for detection of MMP2 and MMP9. Statistical analysis of the semi-quantitative analysis of immunohistochemistry was performed using chi-square test (α = 0.05). In the initial periods of AP progression, an increased expression of MMP9 in the TLR2 KO and MyD88 KO mice was observed. In the final periods of AP progression, a reduction of MMP2 expression and an increase of MMP9 expression in the TLR2 KO mice were observed. MMP2 and MMP9 production was modulated for TLR2 and MyD88 during apical periodontitis progression.


Resumo O objetivo deste estudo foi avaliar a expressão de MMP2 e MMP9 durante a progressão da periodontite apical (AP) em camundongos knockout para TLR2 (TLR2 KO) e MyD88 (MyD88 KO) comparados aos camundongos wild type (WT). A AP foi induzida nos primeiros molares inferiores dos camundongos TLR2 KO (n = 18), MyD88 KO (n = 18) e WT (n = 18). Após 7, 21 e 42 dias, os animais foram eutanaziados e as mandíbulas foram dissecadas e submetidas a processamento histotécnico. As lâminas foram coradas por imuno-histoquímica e analisadas para a detecção de MMP2 e MMP9. A análise estatística semi-quantitativa da imuno-histoquímica foi realizada pelo teste qui-quadrado (α = 0,05). Nos períodos iniciais de progressão AP, foi observada uma expressão aumentada de MMP9 nos camundongos TLR2 KO e MyD88 KO. Nos períodos finais de progressão AP, observou-se uma redução da expressão de MMP2 e um aumento da expressão de MMP9 nos camundongos TLR2 KO. A produção de MMP2 e MMP9 foi modulada por TLR2 e MyD88 durante a progressão da periodontite apical.


Subject(s)
Animals , Mice , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Myeloid Differentiation Factor 88/physiology , Periapical Periodontitis/enzymology , Toll-Like Receptor 2/physiology , Disease Progression , Immunohistochemistry , Mice, Inbred C57BL , Mice, Knockout , Myeloid Differentiation Factor 88/genetics , Periapical Periodontitis/metabolism , Periapical Periodontitis/pathology
8.
J. appl. oral sci ; 26: e20170512, 2018. tab, graf
Article in English | LILACS, BBO | ID: biblio-954511

ABSTRACT

Abstract Objective To evaluate and correlate, in the same research, the mRNA expression and the staining of RANK, RANKL, OPG, TLR2 and MyD88 by immunohistochemistry in the apical periodontitis (AP) progression in mice. Material and Methods AP was induced in the lower first molars of thirty-five C57BL/6 mice. They were assigned to four groups according to their euthanasia periods (G0, G7, G21 and G42). The jaws were removed and subjected to histotechnical processing, immunohistochemistry and real-time reverse transcription-PCR (qRT-PCR). Data were analyzed with parametric and nonparametric tests (α=0.05). Results An increase of positive immunoreactivity for RANK, RANKL, OPG, TLR2 and MyD88 was observed over time (p<0.05). The RANKL expression was different between the groups G0 and G42, G21 and G42 (p=0.006), with G42 presenting the higher expression in both comparations. The OPG expression was statistically different between the groups G0 and G7, G7 and G21 and G7 and G42 (p<0.001), with G7 presenting higher expression in all the time points. The TLR2 expression was different between the groups G0 and G42 (p=0.03), with G42 showing the higher expression. The MyD88 expression presented a statistical significant difference between groups G7, G21 and G42 compared with G0 (p=0.01), with G0 presenting the smallest expression in all the comparisons. The Tnfrsf11/Tnfrsf11b (RANKL/OPG) ratio increased with the AP progression (p=0.002). A moderate positive correlation between MyD88 and RANKL (r=0.42; p=0.03) and between MyD88 and TLR2 (r=0.48; p<0.0001) was observed. Conclusion The expression of the RANK, RANKL, OPG, MyD88 and TLR2 proteins as well as the ratio Tnfrsf11/Tnfrsf11b (RANKL/OPG) increased with AP progression. There was also a moderate positive correlation between the expression Myd88-Tnfrsf11 and Tlr2-Myd88, suggesting the relevance of Tlr2-Myd88 in bone loss due to bacterial infection.


Subject(s)
Animals , Male , Periapical Periodontitis/metabolism , RNA, Messenger/analysis , RANK Ligand/analysis , Myeloid Differentiation Factor 88/analysis , Osteoprotegerin/analysis , Periapical Periodontitis/pathology , Reference Values , Immunohistochemistry , Gene Expression , Disease Progression , Reverse Transcriptase Polymerase Chain Reaction , Toll-Like Receptor 2/analysis , Mice, Inbred C57BL
9.
Article in English | WPRIM | ID: wpr-728627

ABSTRACT

The subgranular zone (SGZ) of hippocampal dentate gyrus (HDG) is a primary site of adult neurogenesis. Toll-like receptors (TLRs), are involved in neural system development of Drosophila and innate immune response of mammals. TLR2 is expressed abundantly in neurogenic niches such as adult mammalian hippocampus. It regulates adult hippocampal neurogenesis. However, the role of TLR2 in adult neurogenesis is not well studied in global or focal cerebral ischemia. Therefore, this study aimed to investigate the role of TLR2 in adult neurogenesis after photochemically induced cerebral ischemia. At 7 days after photothrombotic ischemic injury, the number of bromodeoxyuridine (BrdU)-positive cells was increased in both TLR2 knock-out (KO) mice and wild-type (WT) mice. However, the increment rate of BrdU-positive cells was lower in TLR2 KO mice compared to that in WT mice. The number of doublecortin (DCX) and neuronal nuclei (NeuN)-positive cells in HDG was decreased after photothrombotic ischemia in TLR2 KO mice compared to that in WT mice. The survival rate of cells in HDG was decreased in TLR2 KO mice compared to that in WT mice. In contrast, the number of cleaved-caspase 3 (apoptotic marker) and the number of GFAP (glia marker)/BrdU double-positive cells in TLR2 KO mice were higher than that in WT mice. These results suggest that TLR2 can promote adult neurogenesis from neural stem cell of hippocampal dentate gyrus through increasing proliferation, differentiation, and survival from neural stem cells after ischemic injury of the brain.


Subject(s)
Adult , Animals , Brain , Brain Ischemia , Bromodeoxyuridine , Dentate Gyrus , Drosophila , Hippocampus , Humans , Immunity, Innate , Ischemia , Mammals , Mice , Neural Stem Cells , Neurogenesis , Neurons , Survival Rate , Toll-Like Receptor 2 , Toll-Like Receptors
10.
Braz. j. infect. dis ; 21(5): 545-549, Sept.-Oct. 2017. graf
Article in English | LILACS | ID: biblio-1039201

ABSTRACT

Abstract The present study was aimed to identify the underlying mechanisms of improper renal function in Leishmania donovani infection that causes VL. Mice (BALB/c) were infected with L. donovani and different parameters for proteinuria were assessed. The levels of superoxide anion (O2 -), hydrogen peroxide (H2O2), lipid peroxidation (MDA), inflammatory cytokines, and toll-like receptor (TLR) 2 and 4 expression were found significantly elevated at 60th day in these animals and declined at 90th day post infection. However, TGF-β and caspase 3 activities were higher at 90th day in comparison to 60th day post infection. These findings suggested that exacerbated inflammatory conditions correlate with abnormal renal functions in L. donovani infection, which is further augmented by activated TLRs expressions by circulating leishmanial antigens. Further, the increased levels of TGF-β and caspase 3 at 90th day suggested TGF-β mediated apoptotic cell death of renal and other cells during later stages of disease that may eventually result in release of host and parasitic factors in urine during visceral leishmaniasis.


Subject(s)
Animals , Female , Rats , Transforming Growth Factor beta/blood , Toll-Like Receptor 2/blood , Toll-Like Receptor 4/blood , Kidney/parasitology , Kidney/pathology , Leishmaniasis, Visceral/pathology , Leishmania donovani , Apoptosis , Disease Models, Animal , Leishmaniasis, Visceral/blood , Mice, Inbred BALB C
11.
Braz. dent. j ; 28(3): 281-286, May-June 2017. tab, graf
Article in English | LILACS | ID: biblio-888657

ABSTRACT

Abstract The aim of this study was to find the role of TLR2 signaling pathway in reducing osteoclast activity and promoting osteoblast growth by inducing a combination of Aloe vera and cancellous bovine xenograft (XCB) into dental extraction socket. Forty-eight Cavia cobayas were used. They were divided into eight groups (n=6). For control group, their mandibular incisors were extracted and filled with PEG. For treatment groups, they were extracted and filled with XCB, Aloe vera and the combination of Aloe vera and XCB. The first four groups were sacrificed after 7 days and the other groups after 30 days. Immunohistochemistry and histopathology examination were conducted to examine TLR2, TNFa, OPG, collagen-1, and the osteoblast and osteoclast expressions. The expressions of TLR2, OPG and Collagen-1, as well as the number of osteoblast were increased. Meanwhile, the expressions of TNFa and osteoclast were decreased. The study finding was that TLR2 signaling pathway influenced alveolar bone osteogenesis process by reducing osteoclast activity and stimulating osteoblast growth induced by the combination of Aloe vera and XCB.


Resumo O objetivo deste estudo foi investigar o papel da via de sinalização de TLR2 na redução da atividade osteoclástica e na promoção do crescimento de osteoblastos, induzindo uma combinação de Aloe vera e enxerto de osso esponjoso bovino (EOEB) em alvéolo de extração dentária. Quarenta e oito Cavia cobayas foram utilizados e divididos em 8 grupos (n = 6). Para o grupo de controle, seus incisivos mandibulares foram extraídos e preenchidos com polietilenoglicol (PEG). Para grupos de tratamento, os dentes foram extraídos e preenchidos com EOEB, Aloe vera e a combinação de Aloe vera e EOEB. Os primeiros quatro grupos foram sacrificados após 7 dias e os outros grupos após 30 dias. As análises de imunohistoquímica e histopatologia foram realizada para examinar TLR2, TNFa OPG, colágeno-1 e as expressões de osteoblastos e osteoclastos. Houve maior expressão de TLR2, FGF2, OPG e colágeno-1, bem como maior número de osteoblastos. Enquanto isso, a expressão de TNFa e osteoclastos estava diminuída. O principal achado do estudo foi que a via de sinalização de TLR2 influenciou o processo de osteogênese do osso alveolar, reduzindo a atividade dos osteoclastos e estimulando o crescimento de osteoblastos induzido pela combinação de Aloe vera e EOEB.


Subject(s)
Animals , Male , Cattle , Aloe , Alveolar Process/growth & development , Cancellous Bone/transplantation , Osteogenesis , Signal Transduction , Toll-Like Receptor 2/metabolism , Collagen Type I/metabolism , Guinea Pigs , Heterografts , Osteoprotegerin/metabolism , Tooth Socket , Tumor Necrosis Factor-alpha/metabolism
12.
Mem. Inst. Oswaldo Cruz ; 112(4): 260-268, Apr. 2017. tab, graf
Article in English | LILACS | ID: biblio-841779

ABSTRACT

BACKGROUND Leprosy or hansen’s disease is a spectral disease whose clinical forms mostly depends on host’s immune and genetic factors. Different Toll-like receptors (TLR) variants have been described associated with leprosy, but with some lack of replication across different populations. OBJECTIVES To evaluate the role of polymorphisms in genes TLR1, TLR2 and TLR4 and susceptibility to leprosy in a genetic case control study; to verify the association between genotypes of these markers and the immunological profile in the serum of patients with leprosy. METHODS Pre-designed TaqMan® assays were used to genotype markers at TLR1 (rs4833095, rs5743551), TLR2 (rs7656411, rs3804099) and TLR4 (rs1927914, rs1927911). A panel of cytokines and chemokines was accessed by enzime-linked immunosorbent assay (ELISA) test in the serum of a subgroup of patients with and without leprosy reactions. FINDINGS Our results show an association between the T allele of rs3804099 at the TLR2 gene and increased risk for leprosy per se [Odds ratio (OR) = 1.296, p = 0,022]. In addition, evaluating the association between different genotypes of the TLR1, 2 and 4 markers and cytokine/chemokine serological levels, IL-17 appears as an immunological marker regulated by the polymorphism of the three TLR genes evaluated, whereas different TLR1 genotypes were associated with differential production of IL-12p40 and MCP-1(CCL2). Furthermore, other relevant serum markers such as CXCL-10 and IL-6 seemed to be regulated by TLR2 variants and IL-1β was related to TLR4 genotypes. MAIN CONCLUSIONS All together our data points that the tested TLR markers may have a regulatory role in the immunity against Mycobacterium leprae, by driving the host’s production of key cytokines and chemokines involved in the pathogenesis of this disease.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Chemokines/immunology , Chemokines/blood , Toll-Like Receptor 1/genetics , Toll-Like Receptor 2/genetics , Toll-Like Receptor 4/genetics , Leprosy/genetics , Leprosy/immunology , Case-Control Studies , Polymorphism, Single Nucleotide , Alleles , Enzyme-Linked Immunospot Assay , Genotype
13.
AJMB-Avicenna Journal of Medical Biotechnology. 2017; 9 (1): 8-12
in English | IMEMR | ID: emr-185806

ABSTRACT

Background: M2000 is a newly designed and safe Non-Steroidal Anti-Inflammatory Drug [NSAID]. The aim of this study was to assess the effects of M2000 on expression levels of Suppressor of Cytokine Signaling-1 [SOCS-1] and Src Homology-2 domain containing inositol-5'-phosphatase 1 [SHIP1] proteins via Toll-Like Receptor [TLR] 2/microRNA-155 pathway


Methods: HEK293 TLR2 cell line and Peripheral Blood Mononuclear Cells [PBMCs] were treated by different concentrations of M2000 in MTT assay. RNA was extracted by miRN easy Mini kit. Then, cDNA was synthesized and the expression levels of SOCS1, SHIP1 and miRNA155 were evaluated by Quantitative Real time PCR


Results: Our results showed that M2000 significantly increased the expression levels of SOCS1 and SHIP-1 in Lipopolysachride [LPS]-treated and non-treated cells. Moreover, M2000 decreased expression level of miR-155 in LPS treated PBMCs


Conclusion: M2000 can be used as NSAID in LPS induced inflammation and decrease inflammatory cytokines production by targeting SOCS1, SHIP1 and miR-155 in autoimmune and inflammatory diseases


Subject(s)
Humans , Toll-Like Receptor 2/drug effects , MicroRNAs/drug effects , Suppressor of Cytokine Signaling 1 Protein/drug effects , src Homology Domains , Iran
14.
Braz. oral res. (Online) ; 31: e63, 2017. graf
Article in English | LILACS | ID: biblio-952122

ABSTRACT

Abstract This study was conducted to investigate the roles of different Toll-like receptor (TLR) signaling in Porphyromonas gingivalis (P. gingivalis)-induced and ligature-induced experimental periodontal bone resorption in mice. Wild-type (WT), TLR2 knockout (KO), TLR4KO, and TLR2&4 KO mice with C57/BL6 background were divided into three groups: control, P. gingivalis infection, and ligation. Live P. gingivalis or silk ligatures were placed in the sulcus around maxillary second molars over a 2-week period. Images were captured by digital stereomicroscopy, and the bone resorption area was measured with ImageJ software. The protein expression level of gingival RANKL was measured by ELISA. The gingival mRNA levels of RANKL, IL-1β, TNF-α, and IL-10 were detected by RT-qPCR. The results showed that P. gingivalis induced significant periodontal bone resorption in WT mice and TLR2 KO mice but not in TLR4 KO mice or TLR2&4 KO mice. For all four types of mice, ligation induced significant bone loss compared with that in control groups, and this bone loss was significantly higher than that in the P. gingivalis infection group. RANKL protein expression was significantly increased in the ligation group compared with that in the control group for all four types of mice, and in the P. gingivalis infection group of WT, TLR2 KO, and TLR4 KO mice. Expression patterns of RANKL, IL-1β, TNF-α, and IL-10 mRNA were different in the P. gingivalis infection group and the ligation group in different types of mice. In summary, P. gingivalis-induced periodontal bone resorption is TLR4-dependent, whereas ligation-induced periodontal bone resorption is neither TLR2- nor TLR4-dependent.


Subject(s)
Animals , Periodontitis/microbiology , Alveolar Bone Loss/etiology , Porphyromonas gingivalis/pathogenicity , Disease Models, Animal , Toll-Like Receptor 2/physiology , Toll-Like Receptor 2/genetics , Toll-Like Receptor 4/physiology , Time Factors , Enzyme-Linked Immunosorbent Assay , Reproducibility of Results , Alveolar Bone Loss/microbiology , Tumor Necrosis Factor-alpha/metabolism , Interleukin-10/metabolism , Mice, Knockout , Toll-Like Receptor 2/analysis , Toll-Like Receptor 4/analysis , Toll-Like Receptor 4/genetics , Interleukin-1beta/metabolism , Receptor Activator of Nuclear Factor-kappa B/metabolism , Real-Time Polymerase Chain Reaction , Ligation , Metabolism , Mice, Inbred C57BL
15.
Experimental Neurobiology ; : 186-194, 2017.
Article in English | WPRIM | ID: wpr-22196

ABSTRACT

Despite paramount clinical significance of white matter stroke, there is a paucity of researches on the pathomechanism of ischemic white matter damage and accompanying oligodendrocyte (OL) death. Therefore, a large gap exists between clinical needs and laboratory researches in this disease entity. Recent works have started to elucidate cellular and molecular basis of white matter injury under ischemic stress. In this paper, we briefly introduce white matter stroke from a clinical point of view and review pathophysiology of ischemic white matter injury characterized by OL death and demyelination. We present a series of evidence that Toll-like receptor 2 (TLR2), one of the membranous pattern recognition receptors, plays a cell-autonomous protective role in ischemic OL death and ensuing demyelination. Moreover, we also discuss our recent findings that its endogenous ligand, high-mobility group box 1 (HMGB1), is released from dying OLs and exerts autocrine trophic effects on OLs and myelin sheath under ischemic condition. We propose that modulation of TLR2 and its endogenous ligand HMGB1 can be a novel therapeutic target for ischemic white matter disease.


Subject(s)
Demyelinating Diseases , HMGB1 Protein , Ischemia , Leukoencephalopathies , Myelin Sheath , Oligodendroglia , Receptors, Pattern Recognition , Stroke , Toll-Like Receptor 2 , Toll-Like Receptors , White Matter
16.
São Paulo; s.n; s.n; 2017. 137 p. tab, ilus, graf.
Thesis in Portuguese | LILACS | ID: biblio-878456

ABSTRACT

A esporotricose caracteriza-se como uma micose subcutânea causada por fungos dimórficos do gênero Sporothrix, capazes de acometer o homem e uma grande variedade de animais, dentre eles os felinos. A princípio, Sporothrix schenckii era a única espécie conhecida como responsável pela esporotricose. Após estudos genotípicos e fenotípicos de isolados ambientais, clínicos humanos e animais, verificou-se alta variabilidade entre os isolados e estabeleceu-se a existência de um Complexo Sporothrix. Dentro deste, a maior causadora de surtos epidêmicos, justificada por uma maior virulência e capacidade de evasão da resposta imune, é a espécie Sporothrix brasiliensis. Nesse sentido, dada a ausência de estudos direcionados a está espécie, objetivou-se avaliar a importância de receptores Toll like-2 (TLR-2) e Toll like-4 (TLR-4) na infecção por S. brasiliensis. Além disso, utilizando técnicas de proteômica, procurou-se elucidar proteínas diferencialmente expressas em S. brasiliensis quando comparado à espécie S. schenckii. Para avaliação da resposta imune utilizaram-se modelos in vitro e in vivo de infecção, e para a investigação das proteínas diferencialmente expressas, utilizou-se a técnica de proteômica Bottom-up. A investigação da resposta imune in vitro mostrou a dependência dos receptores TLR-2 e TLR-4 no desencadeamento da resposta imune. Os ensaios in vivo mostraram a importância desses receptores no controle da infecção e dependência dos mesmos na produção de citocinas, principalmente nos primeiros 14 dias de infecção. Na ausência do receptor TLR-2, houve a polarização de resposta Th17 na tentativa de controle da infecção. Quando avaliadas as diferenças entre as espécies S. brasiliensis e S. schenckii, em termos de proteínas expressas, verificou-se que S. brasiliensis expressa diferencialmente 60 proteínas. Dentre essas, 9 são relatadas na literatura, como importantes na virulência e escape imunológico dos principais fungos de importância médica. Os resultados encontrados no presente trabalho permitem concluir que reconhecimento de S. brasiliensis é dependente dos receptores TLR-2 e TLR-4. Estudos que investiguem a utilização de outras vias de sinalização como mecanismos compensatórios, bem como, o sinergismo desses receptores no contexto da infecção por S. brasiliensis são fundamentais na compreensão da fisiopatologia dessa doença. No que tange a caracterização proteica, estudos com mutantes para cada uma das proteínas descritas nesse trabalho devem ser avaliados


Sporotrichosis is a subcutaneous mycosis caused by dimorphic fungi of the genus Sporothrix that affects humans and animals, predominantelly felines. Inicially, Sporothrix schenckii was the only specie associated to sporotrichosis. However, after genotypic and phenotypic studies of human and animal clinical isolates, a high variability among the isolates was found and was concluded the existence of a complex: the Sporothrix Complex. Inside the Sporothrix complex, the major cause of epidemic outbreaks, justified by a greater virulence and ability to evade the immune system, is Sporothrix brasiliensis. Concerning this, the absence of studies directed to this specific specie, the aim was to evaluate the importance of Toll like receptor-2 (TLR-2) and Toll like receptor-4 (TLR-4) during S. brasiliensis infection. In addition, was look using proteomics techniques, the proteins differentially expressed in S. brasiliensis when compared to S. schenckii. To evaluate the immune response, in vitro and in vivo tecniques were used, and for the investigation of differentially expressed proteins, the Bottom-up proteomics technique was used. The investigation of the in vitro immune response showed the dependence of TLR-2 and TLR-4 receptors on phagocytosis and the production of inflammatory mediators, such as cytokines and NO. In vivo assays showed the importance of these receptors to control the infection and their dependence on cytokine production during the first 14 days of infection. In the absence of the TLR-2 receptor, the Th17 response was polarized in an attempt to control the infection. Evaluating the differences between S. brasiliensis and S. schenckii, in terms of expressed proteins, it was verified that S. brasiliensis differentially expressed 60 proteins. Among these, 9 are reported in the literature, as important in the virulence and immune evasion among the most important medical fungi. The results found in the present study allow to conclude that S. brasiliensis recognition is dependent on TLR-2 and TLR-4 receptors. Studies investigating the use of other signaling pathways as compensatory mechanisms, as well as the synergism of these receptors in the context of S. brasiliensis infection, are fundamental to understand the pathophysiology of this disease. Regarding the protein characterization, studies with mutants for each of the proteins described in this work should be evaluated


Subject(s)
Sporothrix/immunology , Virulence/immunology , Proteomics/methods , Sporotrichosis/classification , Toll-Like Receptor 2/analysis , Toll-Like Receptor 4/analysis
17.
Article in English | WPRIM | ID: wpr-116879

ABSTRACT

PURPOSE: Activation of Toll-like receptor 2 (TLR2) present on circulating monocytes in patients with Kawasaki disease (KD) can lead to the production of proinflammatory cytokines and interleukin-10 (IL-10). We aimed to determine the association of the frequency of circulating TLR2+/CD14+ monocytes (FTLR2%) with the outcomes of KD, as well as to compare FTLR2% to the usefulness of sIL-10. METHODS: The FTLR2% in patients with KD was measured by flow cytometry. Serum levels of IL-10 (sIL-10) were determined in 31 patients with KD before the initial treatment with intravenous immunoglobulin (IVIG) and in 21 febrile controls by using enzyme-linked immunosorbent assay. Patients were classified as having coronary artery lesions (CALs) based on the maximal internal diameters of the proximal right coronary artery and proximal left anterior descending coronary artery one month after the initial diagnosis. RESULTS: We found that FTLR2% greater than 92.62% predicted CALs with 80% sensitivity and 68.4% specificity, whereas FTLR2% more than 94.61% predicted IVIG resistance with 66.7% sensitivity and 71.4% specificity. Moreover, sIL-10 more than 15.52 pg/mL predicted CALs and IVIG resistance with 40% and 66.7% sensitivity, respectively, and 73.7% and 76.2% specificity, respectively. CONCLUSION: We showed that measuring FTLR2% before the initial treatment could be useful in predicting CAL development with better sensitivity than sIL-10 and with results comparable to sIL-10 results for the prediction of IVIG resistance in patients with KD. However, further studies are necessary to validate FTLR2% as a marker of prognosis and severity of KD.


Subject(s)
Coronary Vessels , Cytokines , Diagnosis , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Humans , Immunoglobulins , Immunoglobulins, Intravenous , Interleukin-10 , Interleukins , Monocytes , Mucocutaneous Lymph Node Syndrome , Prognosis , Sensitivity and Specificity , Toll-Like Receptor 2 , Toll-Like Receptors
18.
Article in English | WPRIM | ID: wpr-219680

ABSTRACT

PURPOSE: Chronic rhinosinusitis with nasal polyps is a chronic inflammatory disease with markedly increased eosinophils, Th2-type lymphocytes, fibroblasts, and goblet cells. Fungi are commonly associated with airway inflammatory diseases, and thymic stromal lymphopoietin (TSLP) is important in the development of Th2 inflammatory responses. The aim of this study was to investigate the interaction between airborne fungi and nasal fibroblasts in TSLP mRNA and protein expression. METHODS: Inferior turbinate and nasal polyp fibroblasts were stimulated with Alternaria and Aspergillus, respectively, for 48 hours, and TSLP mRNA and protein expressions were measured. The reverse transcriptase polymerase chain reaction was performed for the Toll-like receptor (TLR) mRNA expression of the nasal fibroblasts. To determine the role of TLR in the induction of TSLP, the fibroblasts were transfected with siRNA against TLR2 and TLR5. RESULTS: Alternaria induced TSLP mRNA and protein expression in both inferior turbinate and nasal polyp fibroblasts. The nasal polyp fibroblasts responded more strongly to the fungi. TLR2 and TLR5 mRNA expressions were significantly increased with fungal stimulation and TSLP production was significantly inhibited by siRNA against TLR2. CONCLUSIONS: The results of this study show that TSLP expression could be induced in nasal fibroblasts by exposure to Alternaria and that TLR2 may be involved in the process. The promotion of TSLP production in nasal fibroblasts by airborne fungi may facilitate the development or exacerbation of Th2-type nasal inflammation, especially in CRS with nasal polyps.


Subject(s)
Alternaria , Aspergillus , Eosinophils , Fibroblasts , Fungi , Goblet Cells , Inflammation , Lymphocytes , Nasal Polyps , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger , RNA, Small Interfering , Toll-Like Receptor 2 , Toll-Like Receptors , Turbinates
19.
Article in English | WPRIM | ID: wpr-32083

ABSTRACT

Chronic/cyclic neutropenia, leukocyte adhesion deficiency syndrome, Papillon-Lefèvre syndrome, and Chédiak-Higashi syndrome are associated with severe periodontitis, suggesting the importance of neutrophils in the maintenance of periodontal health. Various Toll-like receptor (TLR) ligands are known to stimulate neutrophil function, including FcR-mediated phagocytosis. In the present study, the effect of TLR2 activation on the non-opsonic phagocytosis of oral bacteria and concomitant production of reactive oxygen species (ROS) by human neutrophils was evaluated. Neutrophils isolated from peripheral blood were incubated with Streptococcus sanguinis or Porphyromonas gingivalis in the presence of various concentrations of Pam3CSK4, a synthetic TLR2 ligand, and analyzed for phagocytosis and ROS production by flow cytometry and chemiluminescence, respectively. Pam3CSK4 significantly increased the phagocytosis of both bacterial species in a dose-dependent manner. However, the enhancing effect was greater for S. sanguinis than for P. gingivalis. Pam3CSK4 alone induced ROS production in neutrophils and also increased concomitant ROS production induced by bacteria. Interestingly, incubation with P. gingivalis and Pam3CSK4 decreased the amounts of ROS, as compared to Pam3CSK4 alone, indicating the possibility that P. gingivalis survives within neutrophils. However, neutrophils efficiently killed phagocytosed bacteria of both species despite the absence of Pam3CSK4. Although P. gingivalis is poorly phagocytosed even by the TLR2-activated neutrophils, TLR2 activation of neutrophils may help to reduce the colonization of P. gingivalis by efficiently eliminating S. sanguinis , an early colonizer, in subgingival biofilm.


Subject(s)
Bacteria , Biofilms , Colon , Flow Cytometry , Humans , Leukocyte-Adhesion Deficiency Syndrome , Ligands , Luminescence , Neutropenia , Neutrophils , Periodontitis , Phagocytosis , Porphyromonas gingivalis , Reactive Oxygen Species , Streptococcus , Toll-Like Receptor 2 , Toll-Like Receptors
20.
Article in English | WPRIM | ID: wpr-35998

ABSTRACT

BACKGROUND/AIMS: There has been controversy about the role of Toll-like receptor 2 (TLR2) in renal injury following ureteric obstruction. Although inhibition of the renin angiotensin system (RAS) reduces TLR2 expression in mice, the exact relationship between TLR2 and RAS is not known. The aim of this study was to determine whether the RAS modulates TLR2. METHODS: We used 8-week-old male wild type (WT) and TLR2-knockout (KO) mice on a C57Bl/6 background. Unilateral ureteral obstruction (UUO) was induced by complete ligation of the left ureter. Angiotensin (Ang) II (1,000 ng/kg/min) and the direct renin inhibitor aliskiren (25 mg/kg/day) were administrated to mice using an osmotic minipump. Molecular and histologic evaluations were performed. RESULTS: Ang II infusion increased mRNA expression of TLR2 in WT mouse kidneys (p < 0.05). The expression of renin mRNA in TLR2-KO UUO kidneys was significantly higher than that in WT UUO kidneys (p < 0.05). There were no differences in tissue injury score or mRNA expression of monocyte chemotactic protein 1 (MCP-1), osteopontin (OPN), or transforming growth factor beta (TGF-beta) between TLR2-KO UUO and WT UUO kidneys. However, aliskiren decreased the tissue injury score and mRNA expression of TLR2, MCP-1, OPN, and TGF-beta in WT UUO kidneys (p < 0.05). Aliskiren-treated TLR2-KO UUO kidneys showed less kidney injury than aliskiren-treated WT UUO kidneys. CONCLUSIONS: TLR2 deletion induced activation of the RAS in UUO kidneys. Moreover, inhibition of both RAS and TLR2 had an additive ameliorative effect on UUO injury of the kidney.


Subject(s)
Amides/pharmacology , Angiotensin II/pharmacology , Animals , Disease Models, Animal , Fibrosis , Fumarates/pharmacology , Kidney/drug effects , Male , Mice, Inbred C57BL , Mice, Knockout , Nephritis, Interstitial/genetics , RNA, Messenger/genetics , Renin/antagonists & inhibitors , Renin-Angiotensin System/drug effects , Toll-Like Receptor 2/deficiency , Ureteral Obstruction/drug therapy
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