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1.
Braz. j. med. biol. res ; 54(7): e10579, 2021. tab, graf
Article in English | LILACS | ID: biblio-1249313

ABSTRACT

NOTCH pathway proteins, including the transcriptional factor HES1, play crucial roles in the development of the inner ear by means of the lateral inhibition mechanism, in which supporting cells have their phenotype preserved while they are prevented from becoming hair cells. Genetic manipulation of this pathway has been demonstrated to increase hair cell number. The present study aimed to investigate gene expression effects in hair cells and supporting cells after Hes1-shRNA lentivirus transduction in organotypic cultures of the organ of Corti from postnatal-day-3 mice. Forty-eight hours after in vitro knockdown, Hes1 gene expression was reduced at both mRNA and protein levels. Myo7a (hair cell marker) and Sox2 (progenitor cell marker) mRNA levels also significantly increased. The modulation of gene expression in the organ of Corti upon Hes1 knockdown is consistent with cell phenotypes related to lateral inhibition mechanism interference in the inner ear. The lentivirus-based expression of Hes1-shRNA is a valuable strategy for genetic interference in the organ of Corti and for future evaluation of its efficacy in protocols aiming at the regeneration of hair cells in vivo.


Subject(s)
Animals , Rats , Cochlea , Basic Helix-Loop-Helix Transcription Factors/genetics , Organ of Corti , Cell Differentiation , Receptors, Notch , Transcription Factor HES-1/genetics , Hair Cells, Auditory
2.
Article in English | WPRIM | ID: wpr-880591

ABSTRACT

Osteosarcoma is the most common malignant tumors of bone. Since 1970s, researchers had used chemotherapy drugs to treat osteosarcoma. However, multidrug resistance is a major adverse reaction that affects the efficacy of chemotherapy drugs, leading to the reduced survival rate of osteosarcoma patients. The Notch signaling pathway plays an important role in osteosarcoma proliferation, which affects tumor resistance by reducing intracellular drug accumulation, regulating epithelial-mesenchymal transition, dysregulating microRNA, disrupting the expression of apoptosis genes, and regulating tumor stem cells.


Subject(s)
Bone Neoplasms/drug therapy , Cell Line, Tumor , Cell Proliferation , Humans , Osteosarcoma/drug therapy , Pharmaceutical Preparations , Receptors, Notch/genetics , Signal Transduction
3.
Braz. j. med. biol. res ; 53(11): e9551, 2020. graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS | ID: biblio-1132485

ABSTRACT

The objective of this study was to investigate the effect of Mycobacterium vaccae on Jagged 1 and gamma delta T17 (γδT17) cells in asthmatic mice. An asthma mouse model was established through immunization with ovalbumin (OVA). Gamma-secretase inhibitor (DAPT) was used to block the Notch signaling pathway. M. vaccae was used to treat asthma, and related indicators were measured. Blocking Notch signaling inhibited the production of γδT17 cells and secretion of cytokine interleukin (IL)-17, which was accompanied by a decrease in Jagged1 mRNA and protein expression in the treated asthma group compared with the untreated asthma group. Similarly, treatment with M. vaccae inhibited Jagged1 expression and γδT17 cell production, which was associated with decreased airway inflammation and reactivity. The Notch signaling pathway may play a role in the pathogenesis of asthma through the induction of Jagged1 receptor. On the other hand, the inhibitory effect of M. vaccae on Jagged1 receptor in γδT17 cells could be used for the prevention and treatment of asthma.


Subject(s)
Animals , Rabbits , Signal Transduction , Mycobacterium , Ovalbumin , Receptors, Notch , Jagged-1 Protein
4.
Biol. Res ; 53: 05, 2020. tab, graf
Article in English | LILACS | ID: biblio-1089075

ABSTRACT

BACKGROUND: LincRNAs have been revealed to be tightly associated with various tumorigeneses and cancer development, but the roles of specific lincRNA on tumor-related angiogenesis was hardly studied. Here, we aimed to investigate whether linc-OIP5 in breast cancer cells affects the angiogenesis of HUVECs and whether the linc-OIP5 regulations are involved in angiogenesis-related Notch and Hippo signaling pathways. METHODS: A trans-well system co-cultured HUVECs with linc-OIP5 knockdown breast cancer cell MDA-MB-231 was utilized to study the proliferation, migration and tube formation abilities of HUVECs and alterations of related signaling indicators in breast cancer cells and their conditioned medium through a series of cell and molecular experiments. RESULTS: Overexpressed linc-OIP5, YAP1, and JAG1 were found in breast cancer cell lines MCF7 and MDA-MB-231 and the expression levels of YAP1 and JAG1 were proportional to the breast cancer tissue grades. MDA-MB-231 cells with linc-OIP5 knockdown led to weakened proliferation, migration, and tube formation capacity of co-cultured HUVECs. Besides, linc-OIP5 knockdown in co-cultured MDA-MB-231 cells showed downregulated YAP1 and JAG1 expression, combined with a reduced JAG1 level in conditioned medium. Furthermore, a disrupted DLL4/Notch/NRP1 signaling in co-cultured HUVECs were also discovered under this condition. CONCLUSION: Hence, linc-OIP5 in MDA-MB-231 breast cancer cells may act on the upstream of the YAP1/Notch/NRP1 signaling circuit to affect proliferation, migration, and tube formation of co-cultured HUVECs in a non-cellular direct contact way through JAG1 in conditioned medium. These findings at least partially provide a new angiogenic signaling circuit in breast cancers and suggest linc-OIP5 could be considered as a therapeutic target in angiogenesis of breast cancers.


Subject(s)
Humans , Female , Transcription Factors/metabolism , Breast Neoplasms/pathology , Neuropilin-1/metabolism , Adaptor Proteins, Signal Transducing/metabolism , Receptors, Notch/metabolism , Tumor Microenvironment , Human Umbilical Vein Endothelial Cells/cytology , Breast Neoplasms/metabolism , Immunohistochemistry , Signal Transduction , Blotting, Western , Reverse Transcriptase Polymerase Chain Reaction , Cell Line, Tumor , Real-Time Polymerase Chain Reaction
5.
Article in English | WPRIM | ID: wpr-739669

ABSTRACT

BACKGROUND: Glioblastoma multiforme (GBM) is a highly malignant brain tumor with a worst prognosis of less than one year despite advance treatment facilities. Among various signaling pathway genes displaying genetic modifications, aberrant expression of Notch pathway genes is frequent in GBM offering novel therapeutic targets. Herbal extracts having anticancer properties are used in adjuvant therapy that is safe and affordable as compared to chemotherapeutics. Bacopa monnieri has been used for the development of brain cells because of its neuroprotective properties. Its anticancer properties have shown to be promising in cancer treatment. METHODS: The anticancer properties of Bacoside A, an active and abundant component of Bacopa monnieri was assessed on U-87 MG cell line and its effects on expression of Notch pathway genes were studied. Cell cycle arrest and apoptosis were studied using flow cytometry. Expression of Notch pathway genes comprising of Notch receptors (notch1, notch2, notch3 and notch4), ligands (jagged1 and jagged2), a component of gamma-secretase complex (APH1A) and downstream target (HES1) were evaluated by quantitative real-time PCR. RESULTS: Bacoside A exhibited considerable cytotoxicity on U-87 MG cells inducing cell cycle arrest and apoptosis. Cell cycle analysis revealed a significant arrest of 39.21% cells in sub-G0 phase at 80 µg/mL concentration, increasing to 53.21% at a higher concentration of 100 µg/mL. The fraction of early apoptotic cells in control was low (3.48%) that increased substantially to 31.36% and 41.11% after 80 µg/mL and 100 µg/mL of Bacoside A treatment respectively. Additionally, the expression of notch1 gene decreased after exposure to Bacoside A with a fold change of 0.05, whereas HES1 gene expression was increased by 25 fold. CONCLUSION: These data indicate that Bacoside A has a possible anticancer activity that could be inducing cell cycle arrest and apoptosis through Notch pathway in GBM in vitro.


Subject(s)
Amyloid Precursor Protein Secretases , Apoptosis , Bacopa , Brain , Brain Neoplasms , Cell Cycle , Cell Cycle Checkpoints , Cell Line , Flow Cytometry , Gene Expression , Glioblastoma , Humans , In Vitro Techniques , Ligands , Prognosis , Real-Time Polymerase Chain Reaction , Receptors, Notch
6.
Article in Chinese | WPRIM | ID: wpr-776540

ABSTRACT

OBJECTIVE@#To investigate the effects of Notch signal on hypoxic induction factor (HIF-1α) and autophagy-associated genes Beclin1, LC3I, LC3II in oxygen-glucose deprivation (OGD) induced myocardial cell injury.@*METHODS@#The OGD model was established using hypoxic culture box and hypoglycemic DMEM medium. The cells were divided into normal control group, OGD group, OGD + NC siRNA group, OGD + Notch1 siRNA group and OGD + HIF-1α siRNA group. Western blot was used to detect the interference effects of HIF-1α siRNA and Notch1 siRNA. The effects of Notch1 siRNA and HIF-1α siRNA on the activity of myocardial cells in OGD model were detected by the CCK-8 assay. The effects of Notch1 siRNA and HIF-1α siRNA on autophage-associated genes Beclin1, LC3I and LC3II expression were detected by Western blot.@*RESULTS@#The results of Western blot showed that HIF-1α siRNA could effectively knock down the expression of HIF-1α in myocardial cells in OGD model, and Notch1 siRNA could effectively knock down the expression of Notch1 and HIF-1α in myocardial cells in OGD model. The result of CCK-8 assay showed that Notch1 siRNA and HIF-1α siRNA reduced the activity of myocardial cells in OGD model, and there was no statistical difference between the two groups. Western blot results showed that Notch1 siRNA and HIF-1α siRNA could reduce the expressions of the autophagy-associated genes Beclin1, LC3I and LC3II, and reduce the ratio of LC3II to LC3I at mRNA level.@*CONCLUSION@#Notch1 plays a role in myocardial protection by regulating the expression of HIF-1α to regulate the autophagy in OGD model cells.


Subject(s)
Autophagy , Beclin-1 , Metabolism , Cell Hypoxia , Cells, Cultured , Glucose , Humans , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Microtubule-Associated Proteins , Metabolism , Myocytes, Cardiac , Cell Biology , Pathology , Oxygen , Receptors, Notch , Metabolism , Signal Transduction
7.
Article in Chinese | WPRIM | ID: wpr-775065

ABSTRACT

OBJECTIVE@#To study the expression of molecules associated with the Notch signaling pathway in children with tuberculosis, as well as the role of this pathway in the pathogenesis of tuberculosis in children.@*METHODS@#A total of 62 children who were diagnosed with tuberculosis from June 2017 to December 2018 were enrolled as the case group, and 64 healthy children were enrolled as the healthy control group. Peripheral venous blood samples with a volume of 2 mL were collected, and quantitative real-time PCR was used to measure the mRNA expression levels of the molecules associated with the Notch signaling pathway (receptors Notch1-4, ligands Jagged1/2 and DLL1/3/4, and downstream target genes Hes1 and Hey1) in leukocytes.@*RESULTS@#Compared with the healthy control group, the case group had significant increases in the mRNA expression levels of Notch1, Notch2, and DLL4 in leukocytes (P0.05).@*CONCLUSIONS@#There are significant increases in the mRNA expression of Notch1/2 and DLL4 in children with tuberculosis, while there are no significant changes in the expression of downstream target genes, suggesting that the Notch signaling pathway, which is activated by the interaction between Notch1/2 and DLL4 after Mycobacterium tuberculosis infection, may play a role in childhood tuberculosis by acting on other target genes, and further studies are needed for clarification.


Subject(s)
Child , Humans , Real-Time Polymerase Chain Reaction , Receptors, Notch , Signal Transduction , Tuberculosis
8.
Article in Chinese | WPRIM | ID: wpr-813033

ABSTRACT

Ulcerative colitis is an intestinal inflammatory disease characterized by diarrhea, abdominal pain and purulent stool. Uncontrolled inflammation caused by macrophage hyper-activation is an important cause of ulcerative colitis. Therefore, inhibiting macrophage hyper-activation is an effective way to treat ulcerative colitis. Notch signaling pathway is involved in regulating the immune response of macrophages and promoting inflammation. NF-κB signaling pathway is the "star pathway" involved in inflammation. NLRP3 inflammatory body is involved in the activation of macrophages. Notch, NF-κB and NLRP3 inflammatory bodies constitute the upstream and downstream signal pathways in the existing immune inflammatory diseases. Notch signal pathway can regulate the activation of macrophage via NF-κB/NLRP3 inflammatory body signaling pathway.


Subject(s)
Colitis, Ulcerative , Cytokines , Humans , Macrophage Activation , NF-kappa B , NLR Family, Pyrin Domain-Containing 3 Protein , Receptors, Notch , Signal Transduction
10.
Rev. cuba. obstet. ginecol ; 44(3): 1-17, jul.-set. 2018.
Article in Spanish | LILACS, CUMED | ID: biblio-1093610

ABSTRACT

La vía de señalización Notch desempeña un papel clave para regular el destino celular, crecimiento, proliferación y la muerte celular programada durante el desarrollo de organismos eucariotas. Esta vía está relacionada con una enorme diversidad de procesos del desarrollo y su disfunción está implicada en el origen de muchas malformaciones congénitas. Se realizó una revisión bibliográfica con el objetivo de actualizar la información sobre la vía de señalización Notch y su relación con el origen de diferentes malformaciones congénitas sensibles a la deficiencia materna de ácido fólico y otros micronutrientes. La literatura médica publicada en idiomas español e inglés se recopiló a través de buscadores como PubMed, Medline, Scielo, Lilacs y la biblioteca Cochrane en enero de 2018 usando palabras clave apropiadas. El conocimiento de esta vía de señalización podría ayudar a comprender mejor algunos aspectos de la morfogénesis, ya que, al actuar como un controlador maestro del destino celular, la proliferación, diferenciación y muerte celular programada, ofrece puntos específicos y susceptibles de intervención que posibilitan la prevención de determinadas malformaciones congénitas en el hombre(AU)


Notch signaling pathway plays a key role to regulate cell grow, fates, proliferation and programmed cell death in development of eukaryotic organisms. This pathway is related with an enormous diversity of developmental processes and its dysfunction is implicated in the origin of many congenital malformations. A review was performed to provide updated information on Notch signaling pathway involved in the origin of some congenital malformations related with maternal deficiency of folic acid and other micronutrients. Published medical literature in Spanish and English languages was retrieved from PubMed, Medline, Scielo, Lilacs and the Cochrane Library in January 2018, using appropriate key words. Knowledge about this signaling pathway could help to better understand some topics of morphogenesis, since by acting as a master controller of cell fate, proliferation, differentiation and programmed cell death, it offers susceptible and specific points which make possible to prevent some human congenital malformations(AU)


Subject(s)
Humans , Receptors, Notch/analysis , Receptors, Notch/genetics , Receptors, Notch/therapeutic use , Morphogenesis/genetics , Congenital Abnormalities/genetics
11.
Article in English | WPRIM | ID: wpr-765277

ABSTRACT

OBJECTIVE: Notch receptors are heterodimeric transmembrane proteins that regulate cell fate, such as differentiation, proliferation, and apoptosis. Dysregulated Notch pathway signaling has been observed in glioblastomas, as well as in other human malignancies. Nerve growth factor (NGF) is essential for cell growth and differentiation in the nervous system. Recent reports suggest that NGF stimulates glioblastoma proliferation. However, the relationship between NGF and Notch1 in glioblastomas remains unknown. Therefore, we investigated expression of Notch1 in a glioblastoma cell line (U87-MG), and examined the relationship between NGF and Notch1 signaling. METHODS: We evaluated expression of Notch1 in human glioblastomas and normal brain tissues by immunohistochemical staining. The effect of NGF on glioblastoma cell line (U87-MG) was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. To evaluate the relationship between NGF and Notch1 signaling, Notch1 and Hes1 expression were evaluated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. To confirm the effects of NGF on Notch1 signaling, Notch1 and Hes1 small interfering RNAs (siRNAs) were used. RESULTS: In immunohistochemistry, Notch1 expression was higher in glioblastoma than in normal brain tissue. MTT assay showed that NGF stimulates U87-MG cells in a dose-dependent manner. RT-PCR and Western blot analysis demonstrated that Notch1 and Hes1 expression were increased by NGF in a dose-dependent manner. After transfection with Notch1 and Hes1 siRNAs, there was no significant difference between controls and 100 nM NGF-β, which means that U87-MG cell proliferation was suppressed by Notch1 and Hes1 siRNAs. CONCLUSION: These results indicate that NGF stimulates glioblastoma cell proliferation via Notch1 signaling through Hes 1.


Subject(s)
Apoptosis , Blotting, Western , Brain , Cell Line , Cell Proliferation , Glioblastoma , Humans , Immunohistochemistry , Nerve Growth Factor , Nervous System , Polymerase Chain Reaction , Receptor, Notch1 , Receptors, Notch , Reverse Transcription , RNA, Small Interfering , Transfection
12.
Article in English | WPRIM | ID: wpr-691404

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect of acupuncture on the Notch signaling pathway in rats with traumatic brain injury and to explore the pathogenesis of acupuncture intervention on traumatic brain injury.</p><p><b>METHODS</b>Feeney's freefall epidural impact method was used to establish a traumatic brain injury model in rats; the rats were randomly divided into a normal group, sham operation group, model group and acupuncture group. Acupuncture was performed in the Baihui (DU 20), Shuigou (DU 26), Fengfu (DU 16), Yamen (DU 15) and Hegu (LI 4) acupoints in the rat, and Yamen was punctured via Fengfu. Then, the rats in each group were randomly divided into three subgroups, namely the day 3 subgroup, day 7 subgroup and day 14 subgroup according to treatment duration. The modified neurological severity scores (mNss) method was used to perform neurobehavioral scoring for evaluating the degree of injury in the rats. The hematoxylin-eosin (HE) staining method was used to observe the pathological change in the brain tissue of rats in each group. Real-time fluorescent quantitative polymerase chain reaction (Q-PCR) technology was used to detect changes in the Notch1, Hes1 and Hes5 gene expression levels in the cortex on the injured side. Western blot was used to detect the protein expression changes.</p><p><b>RESULTS</b>One day after modeling, the mNss scores in the model group and in the acupuncture group were significantly higher than those in the normal and sham operation groups (P<0.01) ; there was no statistically significant difference between the normal group and the sham operation group. The scores decreased with increased treatment time, and the scores in the acupuncture group decreased more significantly than those in the model group (P<0.01). The pathological examination by the HE staining method demonstrated that the brain tissue of the rats in the acupuncture and model groups relatively significantly changed. The Notch1 gene expression level in the acupuncture group was significantly higher than the level in all of the other groups (P<0.01) ; the Hes1 and Hes5 gene expression levels were also higher in the acupuncture group. The expression changes of the Notch1 and Hes1 protein were consistent with that of mRNA. In each experimental group, the mNss score and the pathological results by the HE staining method were consistent with the mRNA results.</p><p><b>CONCLUSION</b>Acupuncture could significantly promote high expression levels of Notch1, Hes1 and Hes5 in the brain tissue of traumatic brain injury rats. Therefore, acupuncture might be an important intervention for inducing endogenous stem cell proliferation and for promoting nerve repair.</p>


Subject(s)
Acupuncture Points , Acupuncture Therapy , Animals , Brain Injuries , Genetics , Pathology , Therapeutics , Brain Ischemia , Pathology , Therapeutics , Male , Nerve Regeneration , Genetics , Rats , Rats, Sprague-Dawley , Receptors, Notch , Genetics , Metabolism , Reperfusion Injury , Genetics , Therapeutics , Signal Transduction , Genetics
13.
Article in Chinese | WPRIM | ID: wpr-690947

ABSTRACT

<p><b>OBJECTIVE</b>To express and purify the mouse endothelial cell-targeted recombinant Notch ligand protein mD1R, and to investigate its effect on hematopoiesis after carbon tetrachloride damage.</p><p><b>METHODS</b>PCR was performed to clone and construct the expression vector pET22b(+)-mD1R. The mD1R successfully transformed into E. coli was induced by IPTG, and purified with Ni-beads affinity chromatography. The target protein was detected by SDS-PAGE. The fluorescence-activated cell sorting analysis (FACS), cell adhesion test, immunofluorescence staining and quantitative real-time PCR were employed to detect the endothelial cell-targeted and Notch signaling-activated biological characteristics of mD1R. The carbon tetrachloride mouse model was established to observe the effects of mD1R on the hematopoietic stem cell (HSC), myeloid cells and lymphoid cells by flow cytometry. The LinScal-1c-Kit cells were sorted by magnetic bead, FACS was performed to analyze the cell cycle, and RT-PCR was employed to observe the expression of interleukin (IL)-10.</p><p><b>RESULTS</b>The prokaryotic expression vector was successfully cloned and constructed. The purity and the activity were confirmed in mD1R recombinant protein. The purified mD1R activated the Notch signaling pathway of hematopoietic stem cells in carbon tetrachloride damaged mouse, and internally elevated the number of HSC and long-term HSC to 2.96-fold and 6.18-fold. In addition, mD1R improved the amplification of the myeloid progenitor cells and the myeloid-derived suppressor cells, particularly the granulocyte/monocyte into blood. Mechanistically, the further analyses suggested that Notch pathway could increase the proliferation of HSC and enhance expression of IL-10 after stress injury.</p><p><b>CONCLUSIONS</b>A new and activated recombinant Notch ligand protein has been obtained successfully to communicate hematopoietic stem cells and hematopoietic microenvironment. The Notch- mediated intrinsic hematopoiesis has been regulated by the anti-inflammatory factor after stress injury.</p>


Subject(s)
Animals , Carbon Tetrachloride , Escherichia coli , Hematopoiesis , Hematopoietic Stem Cells , Ligands , Mice , Receptors, Notch , Signal Transduction
14.
Article in English | WPRIM | ID: wpr-788707

ABSTRACT

OBJECTIVE: Notch receptors are heterodimeric transmembrane proteins that regulate cell fate, such as differentiation, proliferation, and apoptosis. Dysregulated Notch pathway signaling has been observed in glioblastomas, as well as in other human malignancies. Nerve growth factor (NGF) is essential for cell growth and differentiation in the nervous system. Recent reports suggest that NGF stimulates glioblastoma proliferation. However, the relationship between NGF and Notch1 in glioblastomas remains unknown. Therefore, we investigated expression of Notch1 in a glioblastoma cell line (U87-MG), and examined the relationship between NGF and Notch1 signaling.METHODS: We evaluated expression of Notch1 in human glioblastomas and normal brain tissues by immunohistochemical staining. The effect of NGF on glioblastoma cell line (U87-MG) was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. To evaluate the relationship between NGF and Notch1 signaling, Notch1 and Hes1 expression were evaluated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. To confirm the effects of NGF on Notch1 signaling, Notch1 and Hes1 small interfering RNAs (siRNAs) were used.RESULTS: In immunohistochemistry, Notch1 expression was higher in glioblastoma than in normal brain tissue. MTT assay showed that NGF stimulates U87-MG cells in a dose-dependent manner. RT-PCR and Western blot analysis demonstrated that Notch1 and Hes1 expression were increased by NGF in a dose-dependent manner. After transfection with Notch1 and Hes1 siRNAs, there was no significant difference between controls and 100 nM NGF-β, which means that U87-MG cell proliferation was suppressed by Notch1 and Hes1 siRNAs.CONCLUSION: These results indicate that NGF stimulates glioblastoma cell proliferation via Notch1 signaling through Hes 1.


Subject(s)
Apoptosis , Blotting, Western , Brain , Cell Line , Cell Proliferation , Glioblastoma , Humans , Immunohistochemistry , Nerve Growth Factor , Nervous System , Polymerase Chain Reaction , Receptor, Notch1 , Receptors, Notch , Reverse Transcription , RNA, Small Interfering , Transfection
15.
Journal of Experimental Hematology ; (6): 1780-1784, 2018.
Article in Chinese | WPRIM | ID: wpr-774386

ABSTRACT

OBJECTIVE@#To investigate the effect of Notch signaling pathways on differentiation of mouse embryonic stem cells(ESC) into haematopoietic stem cells or haematopoietic progenitors cells(HSC/HPC).@*METHODS@#Mouse embryonic stem cells were proliferated in vitro to form embryoid bodies; the differentiation of embryoid bodies should be induced in vitro, the experiments were divided into BE, control, VEGF, DAPT and VEGF-DAPT groups; HSC/HPC ohenotype: CD117D34Sca1 was detected by flow cytometry; the related gene expression was detected by RT-PCR.@*RESULTS@#The number of VEGF-induced HSC/HPC in VEGF group was significantey higher than that in the control and EB group (P<0.05), suggesting that VEGF promotes ESC differentiation to HSC/HPC; the number of DAPT-induced HSC/HPC in DAPT group was significanty higher than that in the Control and EB groups(P<0.05), suggesting that DAPT promotes ESC differentiation to HSC/HPC; the number of VEGF+DAPT-induced HSC/HPC in VEGF-DAPT group was significantly higher than that in VEGF and DAPT groups(P<0.05), suggesting that DAPT and VEGF play a synergistic role to promote differentiation of ESC into HSC/HPC.@*CONCLUSION@#Notch signal pathway inhibits differentiation of ESC into HSC / HPC by VEGF.


Subject(s)
Animals , Cell Differentiation , Hematopoietic Stem Cells , Mice , Mouse Embryonic Stem Cells , Receptors, Notch , Signal Transduction , Vascular Endothelial Growth Factor A
16.
Article in Chinese | WPRIM | ID: wpr-247645

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the clinical features and genetic cause for a family affected with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL).</p><p><b>METHODS</b>Clinical manifestations, neuroimaging, and genetic analysis were performed.</p><p><b>RESULTS</b>The main clinical features have included stroke, emotional disturbance and history of migraine without progressive memory impairment. A positive family history was confirmed. Cranial MRI has revealed multi-infarct lesions and white matter hyperintensity involving bilateral basal ganglia, subcortex and brain stem. All such features were in keeping with the diagnosis of CADASIL. A rare 2182C>T mutation in exon 14 of the NOTCH3 gene was identified in all available cases.</p><p><b>CONCLUSION</b>Both clinical and molecular features suggested that the family has been affected with CADASIL.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Migraine Disorders , Genetics , Receptor, Notch3 , Receptors, Notch , Genetics
17.
Article in English | WPRIM | ID: wpr-238418

ABSTRACT

The relationship between Kruppel-like factor 4 (KLF4) and the Notch pathway was determined to investigate the effect of KLF4 on the activation of hepatic stellate cells and underlying mechanisms. Fifty SPF BALB/c mice were randomly divided into two groups. A liver fibrosis model was established in 25 mice as the experimental group, and the remaining 25 mice served as controls. On the day 0, 7, 14, and 35, liver tissues were removed for immunofluorescent detection. The Notch pathway inhibitor DAPT was added to the primary original hepatic stellate cells, and KLF4 and Notch-associated factor expression was detected by qRT-PCR. Additionally, the hepatic stellate cell line LX-2 was used to establish control and experimental groups, and was cultured in vitro. LX-2 cells in the experimental groups were treated with DAPT and the Notch activator transforming growth factor-beta 1 separately, whereas those in the control group were given isotonic culture medium. After 48 h, KLF4 expression was examined by Western blotting. After transient transfection of LX-2 cells to increase KLF4, the expression of Notch factor was examined. Immunofluorescence analysis showed that, with the aggravation of liver fibrosis, the absorbance (A) values of KLF4 were decreased (day 0: 980.73±153.19; day 7: 1087.99±230.23; day 14: 390.95±93.56; day 35: 245.99±87.34). The expression of Notch pathway- related factors (Notch-1, Notch-2, and Jagged-1) in the hepatic stellate cell membrane was negatively correlated to KLF4 expression. With the increase of KLF4 expression, Notch-2 (0.73±0.13) and Jagged-1 (0.43±0.12) expression decreased, whereas Notch-1 level was not detectable. When the Notch pathway was inhibited, KLF4 levels generally increased (18.12±1.31). Our results indicate that KLF4 expression is negatively correlated to the Notch pathway in hepatic stellate cells, which may provide a reference for the treatment of hepatic fibrosis.


Subject(s)
Animals , Cell Line , Cells, Cultured , Hepatic Stellate Cells , Metabolism , Kruppel-Like Transcription Factors , Genetics , Metabolism , Liver Cirrhosis , Metabolism , Mice , Mice, Inbred BALB C , Receptors, Notch , Metabolism , Signal Transduction , Transforming Growth Factor beta1 , Metabolism
18.
Protein & Cell ; (12): 159-174, 2016.
Article in English | WPRIM | ID: wpr-757142

ABSTRACT

The Notch signaling pathway is conserved from Drosophila to mammals and is critically involved in developmental processes. In the immune system, it has been established that Notch signaling regulates multiple steps of T and B cell development in both central and peripheral lymphoid organs. Relative to the well documented role of Notch signaling in lymphocyte development, less is known about its role in regulating myeloid lineage development and function, especially in the context of acute and chronic inflammation. In this review article, we will describe the evidence accumulated during the recent years to support a key regulatory role of the Notch pathway in innate immune and inflammatory responses and discuss the potential implications of such regulation for pathogenesis and therapy of inflammatory disorders.


Subject(s)
Animals , B-Lymphocytes , Allergy and Immunology , Pathology , Humans , Inflammation , Allergy and Immunology , Pathology , Receptors, Notch , Allergy and Immunology , Signal Transduction , Allergy and Immunology , T-Lymphocytes , Allergy and Immunology , Pathology
19.
Braz. j. oral sci ; 14(2): 135-140, Apr.-June 2015. tab, ilus
Article in English | LILACS | ID: lil-755036

ABSTRACT

Aim: To detect the expression of molecules associated with Notch signaling pathway in stem cells from human exfoliated deciduous teeth (SHED) cultured in specific differentiation medium, namely, keratinocyte growth medium (KGM). Methods:RNA was extracted from SHED harvested on day 1, 3 and 7. RNA was reverse-transcribed to obtain the cDNA and then proceeded with PCR using specific primers for the Notch signaling pathway molecules (Notch1, Jagged-1, Jagged-2 and, Hes1) as well as stem cell marker (Nanog). PCR products were electrophoresed on a 2% agarose gel and stained with SYBR green. Results:Notch-1 was highly expressed in SHED cultured in KGM and showed increase in density as the days progressed, while Jagged-1 showed a decrease. Jagged-2 on the other hand, showed a slight increase on day 3 followed by a decrease on day 7. However, Hes-1 was not expressed in SHED cultured in KGM. Nanog showed expression only on day 3 and gradually increased in expression on day 7. Conclusions:Notch signaling pathway associated molecules; Notch-1, Jagged-1, Jagged-2, and stem cell marker Nanog are expressed in SHED cultured in KGM which may be involved in the differentiation into epithelial-like cells in human dental pulp tissues.


Subject(s)
Humans , Male , Female , Culture Media , Tooth, Deciduous/cytology , Gene Expression , Keratinocytes , Receptors, Notch , Stem Cells
20.
Int. arch. otorhinolaryngol. (Impr.) ; 19(2): 121-123, Apr-Jun/2015.
Article in English | LILACS | ID: lil-747156

ABSTRACT

Introduction A foreign body (FB) is an object or substance foreign to the location where it is found. FBs in the ear, nose, and throat are a common problem frequently encountered in both children and adults. Objective To analyze FBs in terms of type, site, age, and gender distribution and method of removal. Methods A retrospective study was performed in a tertiary care hospital in the central part of Nepal. The study period was from June 2013 to May 2014. The information was obtained from hospital record books. Results A total of 134 patients had FBs in the ear, nose, or throat; 94 were males and 40 were females. Of the 134 patients, 70 (52.23% ) had FB in the ear, 28 (20.89% ) in the nose, and 36 (26.86% ) in the throat. The FB was animate (living) in 28 (40% ) patients with FB in the ear and 1 (3.5% ) patient with FB in the nose, but the FB was inanimate (nonliving) in any patient with FB in the throat, in 42 (60% ) patients with FB in the ear FB, and in 27 (96.4% ) patients with FB of the nose. The FB was removed with or without local anaesthesia (LA) in 98 (73.13% ) patients, and only 36 patients (26.86% ) required general anaesthesia (GA). The most common age group affected was <10 years. Conclusion FBs in the ear and nose were found more frequently in children, and the throat was the most common site of FB in adults and elderly people. Most of the FBs can be easily removed in emergency room or outpatient department. .


Subject(s)
Animals , Humans , Genes, Tumor Suppressor/physiology , Oncogenes/physiology , Receptors, Notch/physiology , Erythrocytes/physiology , Genes, Switch , Hematologic Neoplasms/drug therapy , Hematologic Neoplasms/metabolism , Hematopoiesis/genetics , Hematopoietic Stem Cells/physiology , Megakaryocytes/physiology , Signal Transduction/physiology
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