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This aim of this study was to assess the molecular mechanism of osteoporosis in schizophrenia patients with risperidone use. Here, we investigated the effects of risperidone on cellular proliferation and apoptosis of a preosteoblast cell line, MC3T3-E1. Cell viability and apoptotic rate of MC3T3-E1 were detected by cell counting kit-8 and flow cytometry at a serial dose of risperidone and at different time points, respectively. Bone transformation relevant gene serum osteocalcin (BGP), collagen 1, tumor necrosis factor-α (TNF-α), osteoprotegerin (OPG), and receptor activator of nuclear factor-κB ligand (RANKL) mRNA levels were determined by real-time PCR (qPCR). Their protein expression patterns were evaluated using western blot. The results revealed that risperidone dramatically inhibited MC3T3-E1 cell proliferation in a dose-dependent manner. It also significantly induced MC3T3-E1 cell apoptosis. TNF-α gene and protein levels were greatly enhanced after risperidone treatment. In contrast, BGP, collagen 1, OPG, and RANKL gene and protein levels were markedly downregulated. Our study indicated that risperidone suppressed MC3T3-E1 cell proliferation and induced apoptosis. It also regulated BGP gene and protein expression.
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Animales , Osteoblastos/efectos de los fármacos , Apoptosis/efectos de los fármacos , Risperidona/farmacología , Proliferación Celular/efectos de los fármacos , Osteocalcina/efectos de los fármacos , Línea Celular , Colágeno/efectos de los fármacos , Factor de Necrosis Tumoral alfa/efectos de los fármacos , Receptor Activador del Factor Nuclear kappa-B/efectos de los fármacos , Osteoprotegerina/efectos de los fármacos , Citometría de FlujoRESUMEN
Resumen Introducción: la enfermedad de hígado graso no alcohólico (NAFLD) constituye un problema de salud pública asociado con el síndrome metabólico; su patogénesis implica el inicio de una cascada de señalización bioquímica compleja y su estimulación continua podría consolidar un proceso de fibrogénesis en el tejido. El objetivo del estudio fue analizar expresión de genes implicados en daño hepático, en los procesos iniciales de la lesión en pacientes con NAFLD o con factores de riesgo relacionados a esta patología, en búsqueda de biomarcadores moleculares útiles a la práctica clínica tales como TGF-pi, COL1A2 y MMP20. Metodología: estudio analítico de corte transversal. Se estudiaron características epidemiológicas, bioquímicas, y expresión génica de TGF-pU, COL1A2 y MMP20 en tejido hepático, en individuos con factores de riesgo para NAFLD. Resultados: se incluyeron 83 participantes con factores de riesgo asociados a NAFLD, 22 individuos (26.5%) fueron diagnosticados con NAFLD mediante ultrasonografía. Los factores de riesgo hallados fueron hipertensión arterial (50.6%), obesidad (49.4%), diabetes mellitus (34.9%) y dislipidemia (21.7%). La dislipidemia fue significativamente asociada con el riesgo de desarrollar NAFLD (OR=4; p=0.011). Se encontraron diferencias significativas para colesterol total (p<0.05); y una expresión génica de TGF--31 (con NAFLD p<0.0001 y sin NAFLD p<0.0001 frente al control) y COL1A2 (con NAFLD p=0.002 y sin NAFLD p=0.955 frente al control) con un patrón de expresión creciente a mayor grado de lesión hepática. Conclusión: para concluir, sugerimos activación de las vías de señalización que conducen a fibrogénesis en individuos con factores de riesgo para NAFLD, y mucho más acentuada en pacientes con NAFLD.
Abstract Introduction: nonalcoholic fatty liver disease (NAFLD) is a public health problem associated with the metabolic syndrome; its pathogenesis implies the start of a complex biochemical signaling cascade and its continuous stimulation could consolidate a fibrogenesis process in the tissue. The aim of the study was to analyze expression of genes involved in liver damage in the initial processes of the lesion in patients with NAFLD or with risk factors related to this pathology, in search of molecular biomarkers useful to clinical practice such as TGF--31, COL1A2 and MMP20. Methodology: cross-sectional analytical study. Epidemiological, biochemical, and gene expression characteristics of TGF--31, COL1A2 and MMP20 in liver tissue in individuals with risk factors for NAFLD were studied. Results: 83 participants with risk factors associated to NAFLD were included; 22 individuals (26.5%) were diagnosed with NAFLD by ultrasonography. The risk factors found were hypertension (50.6%), obesity (49.4%), diabetes mellitus (34.9%) and dyslipidemia (21.7%). Dyslipidemia was significantly associated with the risk of developing NAFLD (OR = 4; p = 0.011). Significant differences were found for total cholesterol (p <0.05); and a gene expression of TGF-P1 (with NAFLD p <0.0001 and without NAFLD p <0.0001 versus control) and COL1A2 (with NAFLD p = 0.002 and without NAFLD p = 0.955 versus control) with a pattern of increasing expression at higher degree of liver injury. Conclusion: to conclude, we suggest activation of the signaling pathways that lead to fibrogenesis in individuals with risk factors for NAFLD, and much more accentuated in patients with NAFLD.
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Humanos , Masculino , Femenino , Enfermedad del Hígado Graso no Alcohólico , Factores de Crecimiento Transformadores , Síndrome Metabólico , Colágeno Tipo I , Metaloproteinasas de la Matriz Asociadas a la Membrana , Hígado GrasoRESUMEN
Objective To explore the effects of microwave irradiation on the proliferation of keloid-derived fibroblasts so as to analyze the expression of collagen-1 and the activation of the signaling pathway involved.Methods Cells from a human keloid scar were cultured in vitro and randomly divided into a control group withont any intervention,a 10 mW/cm2 microwave irradiation (10-MI) group and a 20 mW/cm2 microwave irradiation (20-MI) group.Aliquots of the latter 2 groups were irradiated at their corresponding intensities for 5 min,15 min and 30 min.The growth of fibroblasts was evaluated using MTT assay.The expression of collagen-1 and changes in the phosphorylation of protein JNK were detected using western blotting.Results Compared with the control group,no significant differences in the average growth of the keloid-derived fibroblasts were observed in the 10-MI group,but significant differences were observed in the 20-MI group and among the three sub-groups irradiated for different durations.The expression of type 1 collagen was significantly down-regulated after irradiation in a time-dependent manner.After microwave radiation at 20 mW/cm2,JNK was significantly activated compared to the control group at the different time points.Conclusions Microwave irradiation at 20 mW/cm2 can significantly inhibit the proliferation of keloid-derived fibroblasts and the down-regulalion is correlated with the irradiation's duration.It can also significantly inhibit collagen-1 expression and relieve scar formation through activating the JNK signal pathway.
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Objective To investigate hydrogen peroxide (H2O2) and transforming growth factor-β2 (TGF-β2) induced fibronectin (FN), collagen 1 (COL1), nuclear factor (NF)-κB P65 proteins and interlukin (IL)-1βgene expression in human trabecular meshwork cells (HTMCs), and the interventional mechanism of resveratrol (RSV). Methods (1) HTMCs with 70 to 80%confluency were divided into 5 groups. The experimental groups were treated with serum-free medium and with H2O2 at concentrations of 150, 300, 450 and 800μmol/L. The control group was treated with 0μmol/L H2O2. The protein levels of FN, COL1, NF-κB P65 and NF-κB P65 phosphorylation (P-NF-κB P65) were measured by Western blot assay. The expression of IL-1βgene was measured by qPCR. (2) HTMCs were divided into 3 groups. The control group was treated withserum-free medium and without H2O2 and RSV. The H2O2 group was treated with 300μmol/L H2O2. The H2O2+RSV group was treated with 300μmol/L H2O2 and 25μmol/L resveratrol (RSV). The expressions of proteins and genes mentioned above were detected in three groups. NF-κB P65 nuclear translocation was assessed by immunofluorescence technique. (3) HTMCs were divided into 3 groups. The control group was treated with serum-free medium and without TGF-β2 and RSV. The TGF-β2 group was treated with 5μg/L TGF-β2. The TGF-β2+RSV group was treated with 5μg/L TGF-β2 and 25μmol/L RSV. The expressions of proteins and genes mentioned above were detected in three groups. Results (1) Compared with control group, the protein levels of FN and P-NF-κB P65 were significantly increased in 150, 300, 450 and 800μmol/L groups,the expression levels of COL1 protein and IL-1β gene were significantly increased in 300, 450 and 800 μmol/L groups (P <0.05). There were no statistical significances between other indicators. (2) The expression levels of FN, COL1, P-NF-κB P65 proteins and IL-1βgene were significantly higher in H2O2 group than those in control group, and which were significantly lower in H2O2+RSV group than those in H2O2 group. Compared with control group, only the expression of IL-1βgene was decreased in H2O2+RSV group (P < 0.05). NF-κB P65 was only expressed in cytoplasm in control group, while it was expressed in both cytoplasm and nucleus in H2O2 group. Compared with H2O2 group, NF-κB P65 was mainly expressed in cytoplasm. (3) Compared with control group, the expressions of FN, COL1, P-NF-κB P65 proteins and IL-1β gene were significantly increased in TGF-β2 group (P < 0.05). Compared with TGF-β2 group, the indicators mentioned above were significantly decreased in TGF-β2+RSV group (P<0.05). Conclusion H2O2 and TGF-β2 can upregulate the expression of FN, COL1, P-NF-κB P65 proteins and IL-1βgene in HTMCs, which may be involved in the development and progression of glaucoma. RSV can inhibit the influence of H2O2 and TGF-β2 in HTMCs and exert a protective effect on glaucoma.
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BACKGROUND: Genetic suggest that strongest effect is observed in the premenopausal peak bone mass, which become less with age. However, the evaluation of candidate genes polymorphisms has been most frequently done in postmenopausal women and the results have been controversial. Therefore, we studied the possible association of the peak bone mass and candidate for osteoporosis genes polymorphism in premenopausal women. METHODS: The associations between BMD and polymorphisms of the vitamin D receptor (3'-end region by BsmI restriction enzyme and start codon by FokI restriction enzyme), estrogen receptor (by PvuII and XbaI restriction enzyme), and type I collagen 1 (Sp1 binding site by MscI and BalI restriction enzyme) genes were examined in 100 healthy young Korean women who had a peak bone mass (age 20-35 years). Bone mineral densities were measured by dual energy X-ray absorptiometry (DEXA). Dietary calcium intake was also measured using a food frequency questionnaire. RESULTS: The frequencies of the B allele of the vitamin D receptor gene BsmI polymorphism and the X allele in the estrogen receptor gene, XbaI polymorphisms were lower in Koreans than those in Caucasians. The allelic frequencies of the vitamin vitamin D receptor gene FokI polymorphism and the estrogen receptor gene PvuII polymorphism were similar to those of Caucasians. No significant association was found between BMD and the vitamin D receptor genotype according to BsmI or FokI polymorphisms. There was also no significant relation between the PvuII or XbaI polymorphisms of the estrogen receptor gene and BMD. The associations between BMD and cross-genotypes combining the vitamin D receptor gene (BsmI and FokI) and estrogen receptor gene (PvuII and XbaI) polymorphisms were also analyzed. Among the subjects who lacked the Bf haplotype of the vitamin D receptor gene, the BMD of the femoral neck area was significantly higher in subjects lacking Px haplotypes of the estrogen receptor gene than in those having Px haplotype (p < 0.05). When dietary calcium intake was taken into consideration, there were significant differences in BMD according to the cross-genotype in the group having a low calcium intake (< 500 mg/day). The subjects that lacked the Bf and Px haplotypes had a significantly higher BMD in the femoral neck (p < 0.01), Ward's triangle (p < 0.05), and in the trochanteric area (p < 0.05) than those who lacked Bf but a Px haplotype. We did not find a polymorphism in the Sp1 binding site of the type I collagen 1 gene in our subjects. CONCLUSION: These data suggest that a complex interaction of vitamin D and the estrogen receptor gene with the dietary calcium intake, rather than a polymorphism of a single gene, may influence peak bone mass in healthy young Korean women.