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Arq. gastroenterol ; 58(3): 353-358, July-Sept. 2021. tab
Article in English | LILACS | ID: biblio-1345299


ABSTRACT BACKGROUND: The Prex2 protein is a member of the Rac family proteins that belongs to small G proteins with a critical role in cell migration, cell proliferation, and apoptosis through its effects on PI3K cell signaling pathway and phosphatase activity of PTEN protein. The effect of PREX2 gene expression has been shown in some cancer cells. A survey of PREX2 gene expression in gastric antral epithelial cells of gastric cancer patients with Helicobacter pylori various genotypes infection can conduct to better understanding H. pylori infection's carcinogenesis. METHODS: In a case-control study, PREX2 gene expression was evaluated in gastric antral biopsy samples on four groups of patients referred to Sanandaj hospitals, including gastritis with (n=23) and without (n=27) H. pylori infection and gastric cancer with (n=21) and without (n=32) H. pylori infection. Each gastric biopsy sample's total RNA was extracted and cDNA synthesized by using Kits (Takara Company). The PREX2 gene expression was measured using the relative quantitative real-time RT-PCR method and ΔΔCt formula. RESULTS: The PREX2 gene expression increased in gastric antral biopsy samples of gastritis and gastric cancer patients with H. pylori infection (case groups) than patients without H. pylori infection (control groups) 2.38 and 2.27 times, respectively. The patients with H. pylori vacA s1m1 and sabB genotypes infection showed a significant increase of PREX2 gene expression in gastric cancer antral epithelial cells. CONCLUSION: H. pylori vacA s1m1 and sabB genotypes have the positive correlations with PREX2 gene expression in gastric antral epithelial cells of gastritis and gastric cancer patients.

RESUMO CONTEXTO: A proteína Prex2 é membro das proteínas da família Rac que pertencem a pequenas proteínas G com um papel crítico na migração celular, na proliferação celular e na apoptose através de seus efeitos na via de sinalização celular PI3K e atividade fosfatase da proteína PTEN. O efeito da expressão genética PREX2 tem sido mostrada em algumas células cancerosas. Um levantamento da expressão genética PREX2 em células epiteliais antrais gástricas de pacientes infectados com vários genótipos de Helicobacter pylori pode conduzir a um melhor entendimento da carcinogênese da infecção por H. pylori. MÉTODOS: Em estudo de caso-controle, a expressão genética PREX2 foi avaliada em amostras de biópsia antral gástrica em quatro grupos de pacientes encaminhados aos hospitais de Sanandaj, incluindo gastrite com (n=23) e sem (n=27) infecção por H. pylori e de câncer gástrico com (n=21) e sem (n=32) infecção por H. pylori. O RNA total de cada amostra de biópsia gástrica foi extraído e cDNA sintetizado por meio de kits (Takara Company). A expressão genética PREX2 foi medida utilizando-se o método RT-PCR em tempo real quantitativo relativo e a fórmula ΔΔCt. RESULTADOS: A expressão genética PREX2 aumentou em amostras de biópsia antral gástrica de pacientes com gastrite e câncer gástrico com infecção por H. pylori (grupos de casos) em relação aos sem infecção por H. pylori (grupos de controle) 2,38 e 2,27 vezes, respectivamente. Os pacientes com infecção por genótipos H. pylori vacA s1m1 e sabB apresentaram um aumento significativo da expressão genética PREX2 em células epiteliais antrais de câncer gástrico. CONCLUSÃO: Os genótipos H. pylori vacA s1m1 e sabB têm correlações positivas com a expressão genética PREX2 em células epiteliais antrais gástricas de pacientes com câncer gástrico e gastrites.

Humans , Helicobacter Infections , Guanine Nucleotide Exchange Factors/genetics , Gastritis/genetics , Gastritis/microbiology , Case-Control Studies , Helicobacter pylori , Epithelial Cells/metabolism , Epithelial Cells/microbiology , Gastric Mucosa
Frontiers of Medicine ; (4): 252-263, 2021.
Article in English | WPRIM | ID: wpr-880970


An unexpected observation among the COVID-19 pandemic is that smokers constituted only 1.4%-18.5% of hospitalized adults, calling for an urgent investigation to determine the role of smoking in SARS-CoV-2 infection. Here, we show that cigarette smoke extract (CSE) and carcinogen benzo(a)pyrene (BaP) increase ACE2 mRNA but trigger ACE2 protein catabolism. BaP induces an aryl hydrocarbon receptor (AhR)-dependent upregulation of the ubiquitin E3 ligase Skp2 for ACE2 ubiquitination. ACE2 in lung tissues of non-smokers is higher than in smokers, consistent with the findings that tobacco carcinogens downregulate ACE2 in mice. Tobacco carcinogens inhibit SARS-CoV-2 spike protein pseudovirions infection of the cells. Given that tobacco smoke accounts for 8 million deaths including 2.1 million cancer deaths annually and Skp2 is an oncoprotein, tobacco use should not be recommended and cessation plan should be prepared for smokers in COVID-19 pandemic.

Adult , Animals , COVID-19 , Epithelial Cells , Humans , Lung , Mice , Pandemics , Peptidyl-Dipeptidase A , SARS-CoV-2 , Spike Glycoprotein, Coronavirus , Ubiquitin-Protein Ligases/genetics
Article in English | WPRIM | ID: wpr-880648


OBJECTIVES@#To investigate the role of autophagy in oxalate-induced toxicity of human proximal renal tubular epithelial cell (HK-2).@*METHODS@#HK-2 cells were exposed to oxalate (1 mmol/L) for 2 h and 3-methyladenine (3-MA) was used to inhibit autophagy. Then Western blotting was used to measure the expression of autophagy-related protein LC3II. Cell viability and cell apoptosis were measured by MTT assay and flow cytometry assay, respectively.@*RESULTS@#Cytoplasmic vacuolization was observed in HK-2 cells after treating with oxalate for 2 h. However, 3-MA showed no effects on the formation of cytoplasmic vacuolization regardless of the dose at 1 or 5 mmol/L. The expression of LC3II protein was significantly increased in the HK-2 cells in the presence of oxalate (0.62±0.03 vs 0.35±0.02, @*CONCLUSIONS@#Autophagy of HK-2 cells is enhanced by oxalate at the concentration of 1 mmol/L. Inhibition of 3-MA-induced autophagy protects HK-2 cells from the oxalate-induced cytotoxicity.

Apoptosis , Autophagy , Cell Line , Epithelial Cells , Humans , Oxalates/toxicity
Article in English | WPRIM | ID: wpr-878443


OBJECTIVES@#The effect of Vps4b gene mutation on the expressions of cytokeratin 14 (CK14) and proliferating cell nuclear antigen (PCNA) in the Hertwig's epithelial root sheath (HERS) is investigated.@*METHODS@#The bilateral mandibular tissues of mouse on postnatal days 5, 9, 11, 15, and 19 were removed. The mandibular first molar tissue sections were obtained after paraffin embedding. The CK14 and PCNA expressions in the epithelial root sheath of the normal mouse and Vps4b knockout mouse were compared through immunohistochemistry.@*RESULTS@#On postnatal day 5, the normal mouse began to form HERS and had a strong positive PCNA expression in the HERS cells; on postnatal day 9, the HERS structure was continuous, and PCNA was positive in the HERS cells; on postnatal day 11, a small portion of HERS began to break, and PCNA was weakly positive in the HERS cells; on postnatal day 15, HERS continued to fracture; PCNA was weakly and positively expressed in the HERS cells on the root surface; on postnatal day 19, the tooth root reached normal physiological length, and PCNA was positively expressed in the HERS cells of the terminal part. Similar to the normal mouse, the gene knockout mouse also formed a HERS structure on postnatal day 5. However, HERS began to break on postnatal day 9. On postnatal day 19, only a few fragments of HERS were found on the root surface, and the root development was immature. Moreover, the expression intensity of PCNA in the gene knockout mouse was decreased.@*CONCLUSIONS@#The Vps4b gene mutation may change the CK14 and PCNA expressions, leading to abnormal root development.

ATPases Associated with Diverse Cellular Activities , Animals , Endosomal Sorting Complexes Required for Transport , Epithelial Cells , Keratin-14 , Mice , Mice, Knockout , Proliferating Cell Nuclear Antigen , Tooth Root
Article in English | WPRIM | ID: wpr-878359


Objective@#This study aimed to use an air-liquid interface (ALI) exposure system to simulate the inhalation exposure of motorcycle exhaust particulates (MEPs) and then investigate the benchmark dose (BMD) of MEPs by evaluating cell relative viability (CRV) in lung epithelial BEAS-2B cells.@*Methods@#The MEPs dose was characterized by measuring the number concentration (NC), surface area concentration (SAC), and mass concentration (MC). BEAS-2B cells were exposed to MEPs at different concentrations @*Results@#Our results reveal that BMD of NC and SAC were estimated by the best-fitting Hill model, while MC was estimated by Polynomial model. The BMDL for CRV following ALI exposure to MEPs were as follows: 364.2#/cm @*Conclusion@#These results indicate that MEPs exposure

Benchmarking/statistics & numerical data , Bronchi/physiology , Cell Line , Cell Survival/drug effects , Epithelial Cells/physiology , Humans , Motorcycles , Particulate Matter/adverse effects , Vehicle Emissions/analysis
Braz. j. otorhinolaryngol. (Impr.) ; 86(6): 743-747, Nov.-Dec. 2020. tab, graf
Article in English | LILACS | ID: biblio-1142588


Abstract Introduction: Chronic rhinosinusitis with nasal polyps, a prevalent disease affecting around 2% of the world population, is characterized by symptomatic inflammation of the nasal mucosa and impairment of quality of life. Chronic rhinosinusitis with nasal polyps has a multifactorial etiology, involving a dysfunctional host response to environmental factors. Thus, inflammatory models may be useful to shed light on the pathophysiology of this disease. Micronucleus count has been used to screen DNA damage in various tissues. Objective: To investigate the association between frequency of micronucleus in exfoliated cells from the nasal cavity of patients with chronic rhinosinusitis with nasal polyps and disease severity. Methods: This cross-sectional study included 21 patients with chronic rhinosinusitis with nasal polyps and 19 controls without disease. None of the participants were smokers. Results: Mean micronucleus count was 3.690 per 1000 cells (±2.165) in individuals with vs. 1.237 per 1000 cells (±0.806) in controls; (Student's t test = 4.653, p< 0.001). Nasal surgery in the past 5 years and aspirin-exacerbated respiratory disease were not associated with nicronucleus count (p= 0.251). Conclusion: Micronucleus count seems to be linked to chronic rhinosinusitis with nasal polyps, providing a new perspective for the evaluation of this disorder.

Resumo Introdução: A rinossinusite crônica com pólipos nasais, doença prevalente que afeta cerca de 2% da população mundial, é caracterizada por inflamação sintomática da mucosa nasal e comprometimento da qualidade de vida. A rinossinusite crônica com pólipos nasais tem etiologia multifatorial, envolvendo resposta disfuncional do hospedeiro a fatores ambientais. Assim, modelos inflamatórios podem ser úteis para esclarecer a fisiopatologia dessa doença. A contagem de micronúcleos tem sido usada para rastrear danos no DNA em vários tecidos. Objetivo: Investigar a associação entre a frequência de micronúcleos em células esfoliadas da cavidade nasal de pacientes com rinossinusite crônica com pólipos nasais e a gravidade da doença. Método: Estudo transversal que incluiu 21 pacientes com rinossinusite crônica com pólipos nasais e 19 controles sem doença. Nenhum dos participantes era fumante. Resultados: A contagem média de micronúcleos foi de 3,690 por 1.000 células (± 2,165) nos indivíduos doentes e 1,237 por 1.000 células (± 0,806) nos controles (teste t de Student = 4,653; p < 0,001). A cirurgia nasal nos últimos 5 anos e a doença respiratória exacerbada por aspirina não foram associadas à contagem de micronúcleos (p = 0,251). Conclusão: A contagem de micronúcleos parece estar ligada à rinossinusite crônica com pólipos nasais, proporcionando uma nova perspectiva para a avaliação dessa doença.

Humans , Sinusitis/complications , Rhinitis/complications , Nasal Polyps/complications , Quality of Life , Chronic Disease , Cross-Sectional Studies , Epithelial Cells
Rev. MVZ Córdoba ; 25(2): 41-48, mayo-ago. 2020. graf
Article in Spanish | LILACS | ID: biblio-1340772


RESUMEN Objetivo. Describir la influencia del Suero Fetal Bovino (SFB) en la supervivencia, crecimiento y expresión de organelas celulares en las células epiteliales dentales de rata. Materiales y métodos. Cultivos de células epiteliales dentales de rata fueron llevados a cabo a 37°C en una atmosfera húmeda, en ausencia y a una concentración de 10% de SFB. Una evaluación morfológica fue realizada durante la proliferación y confluencia de las células en cultivo. Dobles marcajes por inmunofluorencia fueron efectuados haciendo uso de anticuerpos anti-actina, anti-TOMM20 y anti-LAMP1. Resultados. Se evidenciaron células epiteliales dentales circulares u ovoides con núcleos voluminosos durante la proliferación y confluencias de manera similar en las células cultivas en presencia y ausencia de SFB. La carencia de SFB impactó negativamente la proliferación de las células epiteliales. No fueron observadas alteraciones en la localización de los inmunomarcajes anti-actina, anti-TOMM20 y anti-LAMP1 en las dos condiciones de cultivos experimentales. Conclusiones. La supresión del SFB en el cultivo de células epiteliales dentales de rata disminuyó la supervivencia, proliferación y sugiere no tener un impacto sobre las organelas evaluadas.

ABSTRACT Objective. Describe the influence of Fetal bovine serum (FBS) on the survival, growth and expression of cellular organelles in rat dental epithelial cells. Material and methods. Cell cultures of rat dental epithelial cells were carried out at 37°C in a humid atmosphere, in the absence and at a concentration of 10% FBS. Morphological evaluation was performed during the proliferation and confluence of cell in culture. Double immunofluorescence labels were made using anti-Actin, anti-TOMM20A, and anti-LAMP1 antibodies. Results. Circular or ovoid dental epithelial cells with bulky nuclei were evidenced during proliferation and confluences in a similar manner in culturing cells in the presence and absence of FBS. The lack of FBS negatively impacts the proliferation of epithelial cells. No alterations were observed in the localization of the anti-actin, anti-TOMM20 and anti-LAMP1 immunomarkers in both conditions of experimental cultures. Conclusion. FBS suppression in rat dental epithelial cells decreased survival, proliferation and suggests not having an impact on the organelles evaluated.

Animals , Cattle , Serum Albumin, Bovine , Cattle , Dental Enamel , Epithelial Cells
Rev. colomb. nefrol. (En línea) ; 7(1): 130-134, ene.-jun. 2020. graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1144380


Resumen En pacientes con enfermedad renal se ha reportado la presencia de células renales reactivas, cuyas alteraciones morfológicas severas dificultan su clasificación e interpretación. El conocimiento de las características morfológicas y los patrones de sedimentos en donde se presentan pueden ser de ayuda para su manejo en los departamentos médicos correspondientes. Aquí, nosotros reportamos la presencia de células agrupadas en acinos, con abundante citoplasma, cariomegalia, contornos nucleares irregulares y nucléolos prominentes, acompañados de cilindruria y cuerpos ovales grasos en el sedimento urinario de dos pacientes con diabetes mellitus, las cuales fueron sugestivas de células renales reactivas.

Abstract In patients with kidney disease, the presence of reactive renal cells has been reported. These cells show several morphological alterations that difficult their classification and interpretation. Therefore, the knowledge of its morphological characteristics and sediments patterns where they can be found will helpful for their correct management by medical departments. Here, we reported the presence of renal cells grouped in acinus with abundant cytoplasm, cariomegaly, irregular nuclear contours and prominent nucleoli, accompanied with cilindruria and fatty oval bodies in the urinary sediment of two patients with Diabetes Mellitus, these cells were named as reactive renal cells.

Humans , Male , Female , Renal Insufficiency, Chronic , Patients , Proteinuria , Diabetes Mellitus , Epithelial Cells , Hematuria , Mexico
Arq. bras. med. vet. zootec. (Online) ; 72(3): 926-930, May-June, 2020. ilus, tab, graf
Article in English | ID: biblio-1129601


Papillomatosis is one of the few diseases in which autovaccine therapy is recommended, especially in the case of mass manifestation in animals. It is noted that papillomas are diagnosed in 15.7% of cases in histological diagnosis of oncological diseases. Therefore, this work studied the profile of oncological diseases in dogs and the clinical effectiveness of autovaccine therapy of papillomatosis. The article gives a morphological description of papillomas used as a biomaterial, and also describes the technological methods of manufacturing an autovaccine used to treat animals against papillomatosis. A therapeutic scheme for administering the vaccine was proposed and tested, and the results of the clinical trial are presented, as well as a reflection on some moments of the mechanism of vaccine therapy for papillomatosis.(AU)

A papilomatose é uma das poucas doenças onde se recomenda a terapia com autovacina, principalmente no caso de manifestação em massa em animais. Nota-se que papilomas são diagnosticados em 15,7% dos casos em diagnóstico histológico de doenças oncológicas. Portanto, este trabalho estudou o perfil das doenças oncológicas em cães e a eficácia clínica da terapia com autovacina em papilomatose. O presente artigo fornece descrição morfológica dos papilomas utilizados como biomaterial, além da descrição dos métodos tecnológicos de fabricação de uma autovacina usada para tratar animais contra a papilomatose. Um esquema terapêutico para a administração da vacina foi proposto e testado, e os resultados do ensaio clínico são apresentados, e é feita uma reflexão sobre alguns momentos do mecanismo da terapia de vacina para papilomatose.(AU)

Animals , Dogs , Papilloma/veterinary , Papilloma/epidemiology , Epithelial Cells/pathology , Autovaccines/therapeutic use , Immunotherapy/veterinary
Braz. dent. j ; 31(2): 135-142, Mar.-Apr. 2020. graf
Article in English | LILACS, BBO | ID: biblio-1132288


Abstract Inflammation of periodontal tissues is the consequence of interaction between periodontal pathogens and immune system. This is associated with increased expression of inflammatory cytokines, which may exert destructive effect to the periodontal tissues when released over long period. The aim of this study was to chronologically track the homeostasis of oral keratinocytes following removal of periodontal pathogens. This was done by investigating expression of selected inflammatory markers and integrity of epithelial monolayers in vitro. Rat oral keratinocytes were stimulated with heat-killed Fusobacterium nucleatum and Porphyromonas gingivalis over 7-days then bacteria were washed away and epithelial cells re-cultured for 3-days. Expression of IL-1β, IL-6, and IL-8 was measured by ELISA while transcription of tissue inhibitor of metalloproteinase-1 (TIMP-1) and matrix metalloproteinase -8 (MMP-8) was measured by polymerase chain reaction before and after removal of bacteria. Integrity of epithelial sheet was investigated by using transepithelial electrical resistance. Data showed general downregulation of IL-1b, IL-6, and IL-8 associated with restoring transcription of TIMP-1 and MMP-8 to normal level following removal of bacteria from epithelial cultures. However, expression of IL-8 and MMP-8 remained significantly higher than unstimulated epithelial cells despite withdrawal of F. nucleatum and P. gingivalis respectively from oral keratinocytes cultures. In addition, integrity of epithelial barrier function remained compromised even after removal of P. gingivalis. Results suggest that even after three days following removal of periodontal pathogens, oral keratinocytes sustained persistent upregulation of certain inflammatory markers that could compromise integrity of epithelial barrier function.

Resumo A inflamação dos tecidos periodontais é a consequência da interação entre patógenos periodontais e o sistema imunológico. Isso está associado ao aumento da expressão de citocinas inflamatórias, que podem exercer efeito destrutivo nos tecidos periodontais quando liberadas por um longo período. O objetivo deste estudo foi rastrear cronologicamente a homeostase dos queratinócitos orais após a remoção dos patógenos periodontais. Isto foi feito através da investigação da expressão de marcadores inflamatórios selecionados e da integridade de monocamadas epiteliais in vitro. Os queratinócitos orais de rato foram estimulados com Fusobacterium nucleatum e Porphyromonas gingivalis destruídas pelo calor por 7 dias, depois as bactérias foram lavadas e as células epiteliais foram cultivadas novamente por 3 dias. A expressão de IL-1b, IL-6 e IL-8 foi medida por ELISA, enquanto a transcrição do inibidor tecidual de metaloproteinase-1 (TIMP-1) e matriz metalopeptidase-8 (MMP-8) foi medida por reação em cadeia da polimerase antes e após a remoção de bactérias. A integridade da folha epitelial foi investigada usando resistência elétrica transepitelial. Os dados mostraram uma regulação negativa geral de IL-1b, IL-6 e IL-8 associada à restauração da transcrição de TIMP-1 e MMP-8 para o nível normal após a remoção de bactérias de culturas epiteliais. No entanto, a expressão de IL-8 e MMP-8 permaneceu significativamente maior que as células epiteliais não estimuladas, apesar da retirada de F. nucleatum e P. gingivalis, respectivamente, das culturas de queratinócitos orais. Além disso, a integridade da função da barreira epitelial permaneceu comprometida mesmo após a remoção de P. gingivalis. Os resultados sugerem que, mesmo após três dias após a remoção dos patógenos periodontais, os queratinócitos orais sustentaram uma regulação positiva persistente de certos marcadores inflamatórios que poderiam comprometer a integridade da função da barreira epitelial.

Animals , Rats , Tissue Inhibitor of Metalloproteinase-1 , Epithelial Cells , Fusobacterium nucleatum , Porphyromonas gingivalis , Homeostasis
Article in English | WPRIM | ID: wpr-828976


Objective@#To screen the differentially expressed proteins (DEPs) in human bronchial epithelial cells (HBE) treated with atmospheric fine particulate matter (PM ).@*Methods@#HBE cells were treated with PM samples from Shenzhen and Taiyuan for 24 h. To detect overall protein expression, the Q Exactive mass spectrometer was used. Gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and Perseus software were used to screen DEPs.@*Results@#Overall, 67 DEPs were screened in the Shenzhen sample-treated group, of which 46 were upregulated and 21 were downregulated. In total, 252 DEPs were screened in the Taiyuan sample-treated group, of which 134 were upregulated and 118 were downregulated. KEGG analysis demonstrated that DEPs were mainly enriched in ubiquitin-mediated proteolysis and HIF-1 signal pathways in Shenzhen PM samples-treated group. The GO analysis demonstrated that Shenzhen sample-induced DEPs were mainly involved in the biological process for absorption of various metal ions and cell components. The Taiyuan PM -induced DEPs were mainly involved in biological processes of protein aggregation regulation and molecular function of oxidase activity. Additionally, three important DEPs, including ANXA2, DIABLO, and AIMP1, were screened.@*Conclusion@#Our findings provide a valuable basis for further evaluation of PM -associated carcinogenesis.

Air Pollutants , Bronchi , Metabolism , Computational Biology , Epithelial Cells , Metabolism , Gene Expression , Humans , Mass Spectrometry , Particle Size , Particulate Matter , Proteomics
Article in Chinese | WPRIM | ID: wpr-828634


OBJECTIVE@#To study the expression level of cAMP response element-binding protein (CREB) in children with recurrent wheezing under three years of age and its effect on the expression of the serum orosomucoid 1-like protein 3 (ORMDL3) gene.@*METHODS@#Thirty-six children with recurrent wheezing under three years of age who visited the hospital from June 2017 to June 2019 were selected as the recurrent wheezing group. Twenty-four healthy children from physical examination were selected as the control group. The CREB expression level in peripheral blood was measured by quantitative real-time PCR. Human bronchial epithelial cells (BEAS-2B) were cultured, and dual-luciferase reporter assay and quantitative real-time PCR were used to investigate the effects of overexpression and siRNA interference of CREB on the promoter activity and mRNA expression of the ORMDL3 gene in the BEAS-2B cells.@*RESULTS@#The expression level of CREB in the recurrent wheezing group was significantly higher than that in the control group (P<0.001). In BEAS-2B cells, overexpression of CREB significantly up-regulated the promoter activity and mRNA expression of the ORMDL3 gene (P<0.05), while siRNA interference of CREB significantly reduced the promoter activity and mRNA expression of the ORMDL3 gene (P<0.05).@*CONCLUSIONS@#The expression of CREB is increased in children with recurrent wheezing, and CREB may be involved in the pathogenesis of recurrent wheezing by regulating expression of the ORMDL3 gene.

Child, Preschool , Cyclic AMP Response Element-Binding Protein , Epithelial Cells , Humans , Membrane Proteins , Genetics , Promoter Regions, Genetic , Respiratory Sounds
Article in Chinese | WPRIM | ID: wpr-828366


The aim of this paper was to observe the effect of salvianolic acid B(Sal B) on high-glucose induced renal tubular epithelial-mesenchymal transition(EMT) in rats, and to explore its possible mechanisms of prevention and treatment of diabetic nephropathy. The rat renal tubular epithelial NRK-52 E cells were cultured in vitro. The cells were divided into control group, high glucose group, high glucose+10 μmol·L~(-1)Sal B group(Sal B), the above 3 groups were set at 6, 12, 24 and 48 h for dynamic observation; high glucose+Sal B different concentration(1, 5, 10 μmol·L~(-1)) groups, high glucose+5.0 μmol·L~(-1) pioglitazone group, high glucose+10 μmol·L~(-1)Sal B+5 μmol·L~(-1)GW9662 group. The protein expression levels of PPARγ, PTEN, α-SMA, E-cadherin and PI3 K/Akt signaling molecules were determined by Western blot. The mRNA expression of PPARγ and PTEN were detected by Real-time PCR. The viabi-lity of NRK52 E cells was determined by MTT assay. The results showed that as compared with control group, the mRNA and protein expression levels of PPARγ and PTEN in high glucose group gradually reduced, the protein expression levels of α-SMA and p-Akt~((Thr308))gradually increased, and the protein expression of E-cadherin gradually reduced(P<0.05). As compared with high glucose group, when increases in Sal B doses, the mRNA and protein expression levels of PPARγ, PTEN in high glucose + different concentrations of Sal B groups gradually increased, the protein expression levels of α-SMA and p-Akt~((Thr308)) gradually reduced, and the protein expression of E-cadherin gradually increased(P<0.05), however, the effect of 1 μmol·L~(-1)concentration of Sal B on the expression of PPARγ mRNA and protein and PTEN mRNA was not significantly different. As compared with high glucose group, the mRNA and protein expression levels of PPARγ mRNA(except 6 h) and protein(except 6 h), PTEN mRNA(except 6 h) and protein(except 6, 12 h) kept increasing, the protein expression levels of α-SMA and p-Akt~((Thr308))(except 6 h) continued to reduce, the protein expression of E-cadherin kept increasing in high glucose+10 μmol·L~(-1) Sal B dynamic observation group(P<0.05). As compared with high glucose group, Sal B and the pioglitazone(PIO) can greatly enhance the expression of PPARγ, PTEN at mRNA and protein levels, enhance the expression of E-cadherin at protein levels, and reduce the expression of α-SMA, p-Akt~((Thr308))protein level(P<0.05), there was no significant difference between the two groups. However, the expression levels of PPARγ and PTEN mRNA and protein, E-cadherin, α-SMA and p-Akt(Thr308) protein in the Sal B+GW9662 control group were not statistically significant compared with the high glucose group. The effect of Sal B was blocked by the PPARγ antagonist GW9662. It can be concluded that Sal B can suppress the NRK52 E cells induced by high-glucose EMT. The mechanism may be related to the activation of PPARγ with Sal B, and the up-regulation of PTEN expression, and thereby inhibiting the fibrosis effect of PI3 K/Akt signaling pathway.

Animals , Benzofurans , Epithelial Cells , Epithelial-Mesenchymal Transition , Glucose , Rats , Transforming Growth Factor beta1
Article in Chinese | WPRIM | ID: wpr-828109


OBJECTIVE@#To evaluate the expression of thymidylate synthase (TS) in myoepithelial cells (MECs) of salivary adenoid tissues and explore its clinical significance.@*METHODS@#Immunohistochemical staining EnVision method was used to detect the expression of TS, P63, Calponin, CK5/6 and S-100 in 32 salivary gland specimens, including 10 non-neoplastic and salivary inflammation specimens, 11 mixed tumor specimens, 5 basal cell carcinoma specimens and 6 adenoid cyst carcinoma specimens. The specificity and sensitivity of TS as a specific molecular marker of salivary muscle epithelial cells were evaluated in comparison with P63, Calponin, CK5/6 and S-100.@*RESULTS@#The expression pattern of TS in all the salivary gland tissue specimens was identical with that of p63. TS and P63 both showed strong immunohistochemical expressions in MECs of salivary adenoid tissue specimens. Calponin, CK5/6, and S-100 showed cytoplasmic/membranous expressions in the MECs. In addition, TS exhibited weak or moderate cytoplasmic expression in a few salivary gland epithelial cells, cancer cells and scattered stromal cells, with negative expression in the cell nuclei. The expression of TS in the MECs of all the salivary adenoid specimens was highly consistent with those of P63, Calponin, CK5/6 and S-100 (>0.05) Except for CK5/6 expression in Salivary inflammation and Salivary gland specimens. Kappa>0.75. The specificity and sensitivity of TS as a molecular marker of MECs were both 100%.@*CONCLUSIONS@#TS is a new specific marker of MECs for differential diagnosis of salivary gland tumors.

Adenoids , Biomarkers, Tumor , Carcinoma, Adenoid Cystic , Epithelial Cells , Humans , Salivary Gland Neoplasms , Thymidylate Synthase
Article in English | WPRIM | ID: wpr-827259


BACKGROUND@#We previously demonstrated that continuous exposure to nitrous acid gas (HONO) for 4 weeks, at a concentration of 3.6 parts per million (ppm), induced pulmonary emphysema-like alterations in guinea pigs. In addition, we found that HONO affected asthma symptoms, based on the measurement of respiratory function in rats exposed to 5.8 ppm HONO. This study aimed to investigate the dose-response effects of HONO exposure on the histopathological alterations in the respiratory tract of guinea pigs to determine the lowest observed adverse effect level (LOAEL) of HONO.@*METHODS@#We continuously exposed male Hartley guinea pigs (n = 5) to four different concentrations of HONO (0.0, 0.1, 0.4, and 1.7 ppm) for 4 weeks (24 h/day). We performed histopathological analysis by observing lung tissue samples. We examined samples from three guinea pigs in each group under a light microscope and measured the alveolar mean linear intercept (Lm) and the thickness of the bronchial smooth muscle layer. We further examined samples from two guinea pigs in each group under a scanning electron microscope (SEM) and a transmission electron microscope (TEM).@*RESULTS@#We observed the following dose-dependent changes: pulmonary emphysema-like alterations in the centriacinar regions of alveolar ducts, significant increase in Lm in the 1.7 ppm HONO-exposure group, tendency for hyperplasia and pseudostratification of bronchial epithelial cells, and extension of the bronchial epithelial cells and smooth muscle cells in the alveolar duct regions.@*CONCLUSIONS@#These histopathological findings suggest that the LOAEL of HONO is < 0.1 ppm.

Alveolar Epithelial Cells , Animals , Bronchi , Dose-Response Relationship, Drug , Emphysema , Epithelial Cells , Guinea Pigs , Hyperplasia , Inhalation Exposure , Lung , Pathology , Male , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Myocytes, Smooth Muscle , Nitrous Acid , Toxicity