RESUMO
Porcine ear size is an important characteristic for distinguishing among pig breeds. In a previous genome-wide association study of porcine ear size, LEM domain-containing 3 (LEMD3), methionine sulfoxide reductase B3 (MSRB3), high mobility group AT-hook 2 (HMGA2), and Wnt inhibitory factor 1 (WIF1) were implicated as important candidate genes for ear size. This study investigated the expression levels of four candidate genes for ear size in Erhualian and Large White pigs. Ten Erhualian pigs with large ears and eight Large White pigs with small ears at 60 days of age were examined. The mRNA expression levels of the four candidate genes were quantified by real-time polymerase chain reaction. WIF1 mRNA expression was significantly higher in Large White than in Erhualian pigs (P < 0.05), whereas the expression levels of the other three genes were not significantly different between the two breeds. The protein expression levels of the four genes were analyzed using western blot. WIF1 protein expression was significantly higher in Large White than in Erhualian pigs (P < 0.01), whereas MSRB3 protein expression was significantly higher in Erhualian than in Large White pigs (P < 0.05). There were no significant differences between the two breeds in residual protein expression. These results suggest that WIF1 is the main causal gene for ear size in pigs.
Assuntos
Orelha/crescimento & desenvolvimento , Locos de Características Quantitativas , Suínos/genética , Animais , Animais Endogâmicos , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Perfilação da Expressão Gênica , Estudo de Associação Genômica Ampla , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Metionina Sulfóxido Redutases/genética , Metionina Sulfóxido Redutases/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
Mineral elements in barley (Hordeum vulgare) play an important physiological role in global human health. In this study, quantitative trait loci (QTLs) for concentration of nine mineral elements in barley grain and grass powder were detected in a population of 193 recombinant inbred lines of the barley cross Ziguangmangluoerling x Schooner and the parents. We observed large genetic variation contributing to element concentrations in both grains and grass powder. The mean K, Ca, and Fe concentrations in grass powder were 6.67, 12.00, and 4.58 times that of regenerating barley grains. In grains, 17 QTLs that accounted for 6.36-64.08% of the phenotypic variation in Zn, Mg, Ca, K, Na, Mn, Fe, and P concentrations were identified. In grass powder, seven QTLs were identified; these accounted for 6.03-21.86% of the variation in Ca, Zn, Mg, K, Fe, and Cu concentrations. These QTLs affecting elements in grain and grass powder are so far unreported in barley. To our knowledge, QTLs with pleiotropic effects for three elements were also identified for the first time in barley. The qK1/qMg1/qCa1 region between markers Bmag0211 and GBMS0014 on chromosome 1H was shown to have large additive effects for Mg, Ca, and K concentrations in grains. These additive effects indicated that the high element (Mg, Ca, Zn, Mn, and K) alleles were contributed by Ziguangmangluoerling. These results will further our understanding of the genetic basis of mineral elements and help us develop markers linked with mineral elements for marker-assisted selection breeding in barley.
Assuntos
Hordeum/genética , Minerais/análise , Locos de Características Quantitativas , DNA de Plantas/genética , Grão Comestível/genética , Variação Genética , Melhoramento Vegetal , Seleção GenéticaRESUMO
The purpose of this study was to investigate the effect of the traditional Chinese medicine TanIIA on the viability, invasion, and metastasis of SW480 cells. SW480 cells were treated with TanIIA for 24 h, and MTT assays were performed to determine the effect of TanIIA on cell viability. Transwell transmembrane experiments were applied to test the effect of 1.0 mg/mL TanIIA on SW480 cell invasion and metastasis abilities. Western blotting was performed to determine the expression of the tumor cell metastasis proteins E-cadherin, vimentin, and MMP-9. The cell growth inhibition rates were 0%, 26 ± 4.3%, 43.47 ± 4.0%, 63.0 ± 5.5%, and 76.8 ± 7.8% for treatment with 0, 0.5, 1.0, 2.0, and 5.0 mg/L TanIIA, respectively. The differences in the cell viability inhibitory rates among all groups were statistically significant (P < 0.05). The Transwell assay results indicated that SW620 cell invasion and metastasis abilities were strongly inhibited by 1.0 mg/mL TanII. The western blotting results showed that the expression of E-cadherin was significantly increased and that the expression levels of vimentin and MMP-9 were significantly decreased after treatment with 1.0 mg/mL TanII for 24 h (P < 0.05). Tan II can effectively inhibit the biological activity of colon cancer in vitro and prevent the invasion of colon cancer cells.
Assuntos
Abietanos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Neoplasias do Colo/tratamento farmacológico , Antígenos CD , Caderinas/biossíntese , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/diagnóstico por imagem , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Humanos , Metaloproteinase 9 da Matriz/biossíntese , Invasividade Neoplásica , Metástase Neoplásica , Vimentina/biossínteseRESUMO
Mitogen-activated protein kinase kinase kinase 5 (MAP3K5) is essential for apoptosis, proliferation, differentiation, and immune responses, and is a candidate marker for residual feed intake (RFI) in pig. We cloned the full-length cDNA sequence of porcine MAP3K5 by rapid-amplification of cDNA ends. The 5451-bp gene contains a 5'-untranslated region (UTR) (718 bp), a coding region (3738 bp), and a 3'-UTR (995 bp), and encodes a peptide of 1245 amino acids, which shares 97, 99, 97, 93, 91, and 84% sequence identity with cattle, sheep, human, mouse, chicken, and zebrafish MAP3K5, respectively. The deduced MAP3K5 protein sequence contains two conserved domains: a DUF4071 domain and a protein kinase domain. Phylogenetic analysis showed that porcine MAP3K5 forms a separate branch to vicugna and camel MAP3K5. Tissue expression analysis using real-time quantitative polymerase chain reaction (qRT-PCR) revealed that MAP3K5 was expressed in the heart, liver, spleen, lung, kidney, muscle, fat, pancrea, ileum, and stomach tissues. Copy number variation was detected for porcine MAP3K5 and validated by qRT-PCR. Furthermore, a significant increase in average copy number was detected in the low RFI group when compared to the high RFI group in a Duroc pig population. These results provide useful information regarding the influence of MAP3K5 on RFI in pigs.
Assuntos
MAP Quinase Quinase Quinase 5/genética , Sus scrofa/genética , Sequência de Aminoácidos , Animais , Domínio Catalítico , Clonagem Molecular , Variações do Número de Cópias de DNA , Dosagem de Genes , Expressão Gênica , Interações Hidrofóbicas e Hidrofílicas , MAP Quinase Quinase Quinase 5/química , MAP Quinase Quinase Quinase 5/metabolismo , Modelos Moleculares , Especificidade de Órgãos , Filogenia , Conformação Proteica em alfa-Hélice , Sus scrofa/metabolismoRESUMO
The signal peptide CUB EGF-like domain-containing protein 3 (SCUBE3) gene is a member of SCUBE gene family and plays important roles in bone cell biology and the determination of limb bone length. In this study, the full-length transcript of porcine SCUBE3 was cloned using reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends. The full-length sequence of porcine SCUBE3 cDNA was 4131 base pairs and included 21 exons. The SCUBE3 gene contained a 2895-base pair open reading frame that encoded a peptide of 965 amino acids. Comparison of the deduced amino acid sequences of porcine SCUBE3 with those of human, mouse, zebrafish, and rat showed 96, 95, 73, and 95% identities, respectively. Porcine SCUBE3 mRNA expression levels were highest in the backfat, bone marrow, and cartilage tissues. Copy number variation was detected in porcine SCUBE3 and validated by real-time quantitative polymerase chain reaction. Different copy number variations were present in randomly selected individuals and may, therefore, be a good marker for identifying phenotypic traits. Our findings provide a basis for further investigation of the functions and regulatory mechanisms of SCUBE3 in pigs.
Assuntos
Clonagem Molecular , Variações do Número de Cópias de DNA , Expressão Gênica , Receptores de Superfície Celular/genética , Sus scrofa/metabolismo , Animais , Medula Óssea/metabolismo , Cartilagem/metabolismo , Especificidade de Órgãos , Filogenia , RNA Mensageiro , Receptores de Superfície Celular/química , Receptores de Superfície Celular/metabolismo , Sus scrofa/genéticaRESUMO
Lipopolysaccharide (LPS), the major component of the outer cell wall of Gram-negative bacteria, activates the immune system and threatens the health of livestock and poultry. However, little is known about the genes and pathways involved in the immune response of ducklings to LPS. To elucidate the genes involved in the response of 7-day-old duckling spleens treated with LPS, RNA from LPS-treated and control duckling spleens was analyzed by RNA-Seq. The results showed 11,095 and 10,840 genes with >10 clean reads in the LPS-treated and control groups, respectively. Among these genes, 89 were differentially expressed (log2 ratio ≥ 1, P ≤ 0.01, false discovery rate ≤ 0.001); 67 of these were upregulated and 22 were downregulated in the LPS-treated group compared to the control. GO and GO-rich analysis showed that differentially expressed genes were enriched in 13 functional categories (P < 0.05). Pathway analysis and pathway richness analysis showed that differentially expressed genes were enriched in six pathway categories (P < 0.05). Further analysis showed that some immune system-related signaling pathways, such as the hematopoietic cell lineage, Toll-like receptor signaling pathway, T cell receptor signaling pathway, T cell receptor signaling pathway, complement and coagulation cascades, antigen processing and presentation, and chemokine signaling pathway, are active during the immune response. To confirm the RNA-Seq results, we detected CCL4, LBP, CD71, and STEAP3 expression using real-time PCR analysis, and the results were consistent with the RNA-Seq results. Our results provide new information on the genes involved in the immune response of duckling spleens to LPS.
Assuntos
Patos/genética , Patos/metabolismo , Regulação da Expressão Gênica , Lipopolissacarídeos/metabolismo , Transdução de Sinais , Baço/metabolismo , Animais , Biologia Computacional/métodos , Patos/imunologia , Perfilação da Expressão Gênica , Ontologia Genética , Sequenciamento de Nucleotídeos em Larga Escala , Imunidade/genética , Lipopolissacarídeos/imunologia , Anotação de Sequência Molecular , Reprodutibilidade dos Testes , Baço/imunologiaRESUMO
The expression of transforming growth factor-beta 1 (TGF-ß1) inside the callus cells of diabetic rats and the impact of insulin therapy on its expression and biomechanics was investigated. The rats were randomly divided as follows: an insulin therapy group (IT), a diabetic model group (DM), and a non-diabetic control group (NC). Bone specimens from each group were extracted at different times for immunohistochemical observation of the expression of TGF-ß1. Concurrently, the destruction torque and torsional stiffness were detected at different times. One to four weeks after fracture, TGF-ß1 was widely expressed in fractured callus cells and periosteal proliferating cells, while the expression inside diabetic cells was significantly reduced. The expression of TGF-ß1 decreased over the first 68 weeks, and the mature bone cells never expressed TGF-ß1. The destruction torque (Nm) detected in the 6th week revealed that there was a statistically significant difference between the DM, NC, and IT groups (P < 0.01). In conclusion, TGF-ß1 expression was significantly reduced inside the callus cells of diabetic rats. Insulin therapy increased TGF-ß1 expression inside the callus cells of diabetic rats and improved the biomechanical characteristics of the callus.
Assuntos
Calo Ósseo/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Hipoglicemiantes/farmacologia , Insulina/farmacologia , Fraturas da Tíbia/tratamento farmacológico , Fator de Crescimento Transformador beta1/genética , Animais , Calo Ósseo/metabolismo , Calo Ósseo/patologia , Proliferação de Células , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Consolidação da Fratura/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Dureza , Masculino , Periósteo/efeitos dos fármacos , Periósteo/metabolismo , Periósteo/patologia , Ratos , Ratos Wistar , Fraturas da Tíbia/complicações , Fraturas da Tíbia/genética , Fraturas da Tíbia/patologia , Torque , Fator de Crescimento Transformador beta1/metabolismoRESUMO
This study aims to investigate the expression changes of RANKL, RUNX2, and OPG in rabbit periodontal tissues under orthodontic force and explore its effect on the remodeling of periodontal tissues. A total of 16 specific pathogen-free rabbits were used in this study. The maxillary appliance was worn on the right (experimental) side, and the appliance-free left side was used as the control. Rabbits were sacrificed after 3, 5, 7, and 14 days of treatment. Changes in the expression levels of OPG, RANKL, and RUNX2 in the periodontium were detected using real-time PCR and western blotting methods. The OPG expression levels decreased after 3 to 14 days of treatment, while the expression levels of RANKL and RUNX2 increased after 3 to 14 days. The OPG expression levels decreased while those of RANKL and RUNX2 increased during orthodontic tooth movement, which suggested that they play a role in the osteogenesis process and the reconstruction of periodontal tissue.
Assuntos
Subunidade alfa 1 de Fator de Ligação ao Core/genética , Osteoprotegerina/genética , Periodonto/metabolismo , Ligante RANK/genética , Técnicas de Movimentação Dentária , Animais , Fenômenos Biomecânicos , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Regulação da Expressão Gênica , Maxila/metabolismo , Maxila/patologia , Aparelhos Ortodônticos , Osteogênese/genética , Osteoprotegerina/metabolismo , Periodonto/patologia , Pressão , Ligante RANK/metabolismo , CoelhosRESUMO
Pre-selection for fruit skin color at the seedling stage would be highly advantageous, with marker-assisted selection offering a potential method for apple pre-selection. A and MdMYB1 alleles are allele-specific DNA markers that are potentially associated with apple skin color, and co-segregate with the Rf and Rni loci, respectively. Here, we assessed the potential application of these 2 alleles for marker-assisted breeding across 30 diverse cultivars and 2 apple seedling progenies. The red skin color phenotype was usually associated with the MdMYB1-1 allele and A(1) allele, respectively, while the 2 molecular markers provided approximately 91% predictability in the 'Fuji' x 'Cripps Pink' and 'Fuji' x 'Gala' progenies. The results obtained from the 30 cultivars and 2 progenies were consistent for the 2 molecular markers. Hence, the results supported that Rf and Rni could be located in a gene cluster, or even correspond to alleles of the same gene. Our results are consistent with the hypothesis that red/yellow dimorphism is controlled by a monogenic system, with the presence of the red anthocyanin pigmentation being dominant. In addition, our results supported that the practical utilization of the 2 function markers to efficiently and accurately select red-skinned apple cultivars in apple scion breeding programs.
Assuntos
Frutas/genética , Malus/genética , Pigmentação/genética , Proteínas de Plantas/genética , Seleção Genética , Fatores de Transcrição/genética , Alelos , Cruzamento/métodos , Cor , Cruzamentos Genéticos , DNA de Plantas/análise , DNA de Plantas/genética , Genótipo , Malus/classificação , Fenótipo , Reação em Cadeia da Polimerase , Especificidade da EspécieRESUMO
The aim of this study was to observe the effects of atorvastatin combined with ezetimibe on carotid atherosclerosis in elderly patients with hypercholesterolemia. A total of 84 elderly hypercholesterolemic patients complicated with carotid atherosclerosis were divided into control group (atorvastatin alone) and combined group (atorvastatin combined with ezetimibe) and treated for 12 months. Carotid atherosclerosis-related indicators including blood lipid and high-sensitivity C-reactive protein (hsCRP) were determined before and after treatment. The levels of carotid intima-media thickness (CIMT), serum low density lipoprotein cholesterol (LDL-C) and hsCRP were markedly decreased (P < 0.05) after treatment in the two groups, while the reduction of the levels of CIMT, serum LDL-C and hsCRP was more significant in the combined group (P < 0.01). After treatment, the levels of CIMT, serum LDL-C and hsCRP were distinctly different between combined and control group (P < 0.05). The combination of atorvastatin with ezetimibe could further decrease LDL-C and hsCRP levels and have certain effects on the progression of carotid atherosclerosis in elderly patients with hypercholesterolemia.
Assuntos
Azetidinas/administração & dosagem , Doenças das Artérias Carótidas/tratamento farmacológico , Ácidos Heptanoicos/administração & dosagem , Hipercolesterolemia/tratamento farmacológico , Pirróis/administração & dosagem , Idoso , Atorvastatina , Proteína C-Reativa/metabolismo , Doenças das Artérias Carótidas/sangue , Doenças das Artérias Carótidas/complicações , Doenças das Artérias Carótidas/patologia , Espessura Intima-Media Carotídea , LDL-Colesterol/sangue , Combinação de Medicamentos , Ezetimiba , Feminino , Humanos , Hipercolesterolemia/sangue , Hipercolesterolemia/complicações , Hipercolesterolemia/patologia , MasculinoRESUMO
Liver cirrhosis is one of the most common diseases of Chinese patients. Herein, we report the high expression of a newly identified histone 3 lysine 4 demethylase, retinoblastoma binding protein 2 (RBP2), and its role in liver cirrhosis in humans. The siRNA knockdown of RBP2 expression in hepatic stellate cells (HSCs) reduced levels of α-smooth muscle actin (α-SMA) and vimentin and decreased the proliferation of HSCs; and overexpression of RBP2 increased α-SMA and vimentin levels. Treatment with transforming growth factor ß (TGF-ß) upregulated the expression of RBP2, α-SMA, and vimentin, and the siRNA knockdown of RBP2 expression attenuated TGF-ß-mediated upregulation of α-SMA and vimentin expression and HSC proliferation. Furthermore, RBP2 was highly expressed in cirrhotic rat livers. Therefore, RBP2 may participate in the pathogenesis of liver cirrhosis by regulating the expression of α-SMA and vimentin. RBP2 may be a useful marker for the diagnosis and treatment of liver cirrhosis.
Assuntos
Actinas/metabolismo , Células Estreladas do Fígado/metabolismo , Histona Desmetilases/metabolismo , Cirrose Hepática/metabolismo , Proteína 2 de Ligação ao Retinoblastoma/metabolismo , Vimentina/metabolismo , Animais , Western Blotting , Proliferação de Células , Modelos Animais de Doenças , Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Masculino , RNA Interferente Pequeno/metabolismo , Ratos Wistar , Fator de Crescimento Transformador beta/metabolismoRESUMO
Liver cirrhosis is one of the most common diseases of Chinese patients. Herein, we report the high expression of a newly identified histone 3 lysine 4 demethylase, retinoblastoma binding protein 2 (RBP2), and its role in liver cirrhosis in humans. The siRNA knockdown of RBP2 expression in hepatic stellate cells (HSCs) reduced levels of α-smooth muscle actin (α-SMA) and vimentin and decreased the proliferation of HSCs; and overexpression of RBP2 increased α-SMA and vimentin levels. Treatment with transforming growth factor β (TGF-β) upregulated the expression of RBP2, α-SMA, and vimentin, and the siRNA knockdown of RBP2 expression attenuated TGF-β-mediated upregulation of α-SMA and vimentin expression and HSC proliferation. Furthermore, RBP2 was highly expressed in cirrhotic rat livers. Therefore, RBP2 may participate in the pathogenesis of liver cirrhosis by regulating the expression of α-SMA and vimentin. RBP2 may be a useful marker for the diagnosis and treatment of liver cirrhosis.
Assuntos
Animais , Humanos , Masculino , Actinas/metabolismo , Células Estreladas do Fígado/metabolismo , Histona Desmetilases/metabolismo , Cirrose Hepática/metabolismo , /metabolismo , Vimentina/metabolismo , Western Blotting , Proliferação de Células , Modelos Animais de Doenças , Expressão Gênica , Técnicas de Silenciamento de Genes , Ratos Wistar , RNA Interferente Pequeno/metabolismo , Fator de Crescimento Transformador beta/metabolismoRESUMO
Staphylococcus aureus is an important cause of bloodstream infections worldwide. We examined the prevalence of genes that encode erythromycin ribosome methylase and bacterial toxins in S. aureus collected from bloodstream infections. Sixty different S. aureus isolates were obtained from blood cultures of patients who were admitted to a Teaching Hospital in Tianjin from January 2006 to August 2011. The susceptibility of the isolates to 16 antibiotics was tested. Methicillin-resistant S. aureus (MRSA) was identified using the disk diffusion method with cefoxitin. PCR was used to detect genes that encode the staphylococcal enterotoxins, Panton-Valentine leukocidin, toxic shock syndrome toxin 1 and erythromycin ribosome methylase. Molecular analysis of the MRSA strains was done using pulsed-field gel electrophoresis (PFGE) and staphylococcal cassette chromosome mec (SCCmec) typing. The positivity rates of mecA, ermA, ermB, and ermC in the isolates were 13/60, 10/60, 18/60, and 18/60, respectively. Among the 60 isolates, 30 harbored enterotoxin genes, with sea as the most frequent toxin gene (33%), followed by sec (15%), sed (12%), and seb (5%). The see and tst genes were not found in any of the isolates. The pvl gene was detected in four strains. Eleven MRSA isolates were of the SCCmec type III; two MRSA isolates could not be determined through SCCmec typing. PFGE analysis of the 13 MRSA isolates produced 8 distinct pulsotypes. Virulence genes and erythromycin ribosome methylase genes were highly prevalent in these isolates. The PFGE results demonstrated that the MRSA spread through cloning, mainly involving SCCmec type III.
Assuntos
Genes Bacterianos/genética , Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Toxinas Bacterianas/genética , Técnicas de Tipagem Bacteriana , China , DNA Bacteriano/análise , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Exotoxinas/genética , Frequência do Gene , Hospitais de Ensino , Humanos , Leucocidinas/genética , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Metiltransferases/genética , Testes de Sensibilidade Microbiana , Proteínas de Ligação às Penicilinas , Reação em Cadeia da Polimerase , Especificidade da Espécie , Infecções Estafilocócicas/sangue , Staphylococcus aureus/classificação , Staphylococcus aureus/patogenicidade , Virulência/efeitos dos fármacos , Virulência/genéticaRESUMO
This study was designed to investigate the effect of curcumin (diferuloylmethane) on the proliferation and apoptosis of hepatic stellate cells (HSC). The cell line HSC-T6 (1.25 x 10(5) cells/mL) was incubated with curcumin and HSC proliferation was detected by a methyl thiazolyl tetrazolium colorimetric assay. HSC apoptosis was detected by flow cytometry, transmission electron microscope and agarose gel electrophoresis. HSC proliferation was significantly inhibited in a concentration-dependent manner (10.6 to 63.5 percent) after incubation with 20-100 ìM curcumin, compared with a control group. At 20, 40, and 60 ìM, after 24 h of incubation, curcumin was associated with a significant increase in the number of HSC in the G2/M phase, and a significant decrease in cell numbers in the S phase (P < 0.05). At these concentrations, curcumin was also associated with an increase in the apoptosis index of 15.3 ± 1.9, 26.7 ± 2.8, and 37.6 ± 4.4 percent, respectively, compared to control (1.9 ± 0.6 percent, P < 0.01). At 40 ìM, the curcumin-induced apoptosis index at 12, 24, 36, and 48 h of incubation was 12.0 ± 2.4, 26.7 ± 3.5, 33.8 ± 1.8, and 49.3 ± 1.6 percent, respectively (P < 0.01). In conclusion, curcumin inhibits the in vitro proliferation of HSCs in the G2/M phase of the cell cycle and also induces apoptosis in a concentration- and time-dependent manner. The in vivo effect of curcumin on HSCs requires further investigation.
Assuntos
Animais , Ratos , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Curcumina/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Linhagem Celular , Colorimetria , Eletroforese em Gel de Ágar , Citometria de Fluxo , Células Estreladas do Fígado/patologia , Microscopia Eletrônica de Transmissão , Fatores de TempoRESUMO
This study was designed to investigate the effect of curcumin (diferuloylmethane) on the proliferation and apoptosis of hepatic stellate cells (HSC). The cell line HSC-T6 (1.25 x 10(5) cells/mL) was incubated with curcumin and HSC proliferation was detected by a methyl thiazolyl tetrazolium colorimetric assay. HSC apoptosis was detected by flow cytometry, transmission electron microscope and agarose gel electrophoresis. HSC proliferation was significantly inhibited in a concentration-dependent manner (10.6 to 63.5%) after incubation with 20-100 microM curcumin, compared with a control group. At 20, 40, and 60 microM, after 24 h of incubation, curcumin was associated with a significant increase in the number of HSC in the G2/M phase, and a significant decrease in cell numbers in the S phase (P < 0.05). At these concentrations, curcumin was also associated with an increase in the apoptosis index of 15.3 +/- 1.9, 26.7 +/- 2.8, and 37.6 +/- 4.4%, respectively, compared to control (1.9 +/- 0.6%, P < 0.01). At 40 microM, the curcumin-induced apoptosis index at 12, 24, 36, and 48 h of incubation was 12.0 +/- 2.4, 26.7 +/- 3.5, 33.8 +/- 1.8, and 49.3 +/- 1.6%, respectively (P < 0.01). In conclusion, curcumin inhibits the in vitro proliferation of HSCs in the G2/M phase of the cell cycle and also induces apoptosis in a concentration- and time-dependent manner. The in vivo effect of curcumin on HSCs requires further investigation.