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1.
Chinese Journal of Reparative and Reconstructive Surgery ; (12): 1438-1445, 2020.
Article in Chinese | WPRIM | ID: wpr-856222

ABSTRACT

Objective: To investigate the effects of silencing P75 neurotrophin receptor (P75NTR) and nerve growth factor (NGF) overexpression on the proliferative activity and ectopic osteogenesis ability of bone marrow mesenchymal stem cells (BMSCs) combined with demineralized bone matrix for heterotopic osteogenesis. Methods: BMSCs of Sprague Dawley (SD) rats were cultured and passaged by adherent isolation method. The third generation BMSCs were transfected with lentivirus mediated P75NTR gene silencing (group B), NGF overexpression gene (group C), P75NTR silencing and NGF overexpression double genes (group D), respectively, and untransfected cells as control (group A). After 7 days of transfection, the expression of fluorescent protein of the target gene was observed by fluorescence microscope; cell counting kit 8 method was used to detect the cells activity for 8 days after transfection; the expressions of P75NTR and NGF proteins in each group were detected by Western blot. The adhesion of BMSCs to demineralized bone matrix (DBM) was observed by inverted phase contrast microscope and scanning electron microscope after transfection of p75NTR silencing and NGF overexpression double genes. After transfection, BMSCs and DBM were co-cultured to prepare 4 groups of tissue engineered bone, which were respectively placed in the dorsal subcutaneous tissue of 8-week-old SD rats to construct subcutaneous ectopic osteogenesis model ( n=6). HE staining was performed at 4 and 8 weeks after operation. ALP staining was used to observe the formation of calcium nodules at 8 weeks after operation. The expressions of Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), and osteocalcin (OCN) were detected by real-time fluorescent quantitative PCR. Results: At 7 days after transfection, there was no fluorescence expression in group A, red fluorescence expression was seen in group B, green fluorescence expression in group C, and red-green compound fluorescence expression in group D. The fluorescence expression rate of target gene was about 70%. Western blot detection showed that the relative expression of P75NTR protein in groups A and C was significantly higher than that in groups B and D, and the relative expression of NGF protein in groups C and D was significantly higher than that in groups A and B ( P<0.05). With the passage of time, the cell proliferation activity increased in all groups, especially in group D, which was significantly higher than that in group A at 3-8 days ( P<0.05). The results of inverted phase contrast microscope and scanning electron microscope showed that BMSCs could adhere well to DBM. In the subcutaneous ectopic osteogenesis experiment, HE staining showed that at 4 and 8 weeks after operation, the more bone tissue was formed in group D than in the other 3 groups. ALP staining showed that group D had the highest ALP activity and better osteogenic expression. Compared with group A, the relative expressions of Runx2, ALP, and OCN mRNAs in group D were significantly higher than those in group A ( P<0.05). Conclusion: Silencing P75NTR and NGF overexpression double genes co-transfected BMSCs with DBM to construct tissue engineered bone has good ectopic osteogenic ability. By increasing NGF level and closing P75NTR apoptosis channel, it can not only improve cell activity, but also promote bone tissue regeneration.

2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 174-181, 2019.
Article in Chinese | WPRIM | ID: wpr-801949

ABSTRACT

Objective: To set a criterion for determining whether herbs distribute to kidney meridian from the perspective of tissue expression of protein receptors, so as to provide new ideas and a new method for the quantitative study of meridian tropism. Method: The 9 Yang-tonifying herbs were selected as the training set, and 2 Yang-tonifying herbs were used as the verification set. The TCMSP2.3,PubChem,Uniprot and other database were used to collect the active compounds and targets of traditional Chinese medicine(TCM). The core target proteins of Yang-tonifying herbs were obtained by using the Maximum Similarity Algorithm for TCM in the training set. The THPA database was used to collect expressions of tissues and target proteins. The empirical regression equation was constructed to explore the tissue distribution of the receptors in the training set, and the criterion for determining whether herbs distribute to kidney meridian was established. The criteria model was tested through validation set data. Result: The herb-active ingredient-protein receptor-tissue expression data library was constructed. A total of 39 core target proteins of Yang-tonifying herbs were acquired. The equations in the training set were highly consistent, with no statistical difference (P=0.999 7). The data of the combined training set was finally fitted to a judgment equation. The model was successfully tested with herbs in the validation set. The accuracy of the model was 100%. Conclusion: This study explored a new method for judging whether TCM distributes to kidney meridian, established an effective criterion model and verified the reliability of the new method. It provides a theoretical basis for the modernization of meridian tropism of traditional Chinese medicine, and is of great significance for the rapid development of traditional Chinese medicine.

3.
Rev. Investig. Salud. Univ. Boyacá ; 6(2): 156-179, 2019. graf, esq
Article in Spanish | LILACS | ID: biblio-1100656

ABSTRACT

Introducción. La malaria es un problema de salud pública a nivel mundial y es causada por 5 especies de parásitos apicomplejos del género Plasmodium. La invasión exitosa de los merozoítos al glóbulo rojo es una etapa fundamental en el ciclo de vida del parásito, el cual usa un variado repertorio de ligandos que interactúan de forma específica con receptores presentes en la membrana del glóbulo rojo. Objetivo. Revisar las características moleculares y estructurales de los receptores expresados en la superficie de los glóbulos rojos, implicados en el proceso de invasión del merozoito de Plasmodium falciparum. Método. Revisión descriptiva sobre las características moleculares y estructurales de los receptores de la superficie del glóbulo rojo, los cuales juegan un papel fundamental durante la invasión del merozoíto de Plasmodium falciparum. Esta revisión empezó por la búsqueda de literatura publicada hasta el año 2019 en bases de datos electrónicas, especializadas en la divulgación de investigación biomédica. Se encontraron 127 documentos, de los cuales se seleccionaron 111 y se excluyeron 33 por no cumplir los criterios de inclusión; en total, se analizaron 78 referencias. Conclusión. En esta revisión se resumieron las características moleculares y estructurales de los receptores presentes en el glóbulo rojo importantes en el proceso de invasión del merozoito de P. falciparum. También, se resaltó la importancia de elucidar las diferentes vías de invasión del parásito y así, poder desarrollar alternativas profilácticas o terapéuticas que conduzcan a mitigar o eliminar la malaria


Introduction. Malaria is a public health problem worldwide. It is caused by 5 species of the Apicomplexa genus Plasmodium. The successful invasion of the erythrocyte by Plasmodium merozoites is a critical stage in the life cycle of the parasite, which uses a broad repertoire of ligands that interact in a specific way with receptors expressed on the membrane of the erythrocyte. Objective. To review the molecular and structural characteristics of the receptors expressed on the erythrocyte surface, involved in the process of merozoite invasion by Plasmodium falciparum. Method. Here, we descriptively review of the molecular and structural characteristics of the red blood cell surface receptors, which play a key role during the invasion of Plasmodum falciparum merozoite. To this purpose, we searched the literature published until 2019 in electronic databases specialized in biomedical research. 127 documents were found, of these, 111 were selected, 33 were excluded and 78 references were analyzed. Conclusion. In this review, the molecular and structural characteristics of the receptors expressed on the erythrocytes and important in the process of invasion of P. falciparum merozoites were discussed. With this, we highlight the importance of elucidating the different invasion pathways the parasite, in order to develop prophylactic or therapeutic alternatives that could lead to mitigate or eliminate malaria.


Introdução. A malária é um problema de saúde pública a nível mundial, é causada por 5 espécies de parasitos do Filo Apicomplexa, do gênero Plasmodium. A invasão bem-sucedida de merozoítos nas hemácias, é uma etapa fundamental no ciclo de vida do parasita, que usa um repertório variado de ligandos que interatuam especificamente com receptores presentes na membrana dos glóbulos vermelhos. Objetivo. Revisão descritiva das características moleculares e estruturais dos receptores da superfície dos glóbulos vermelhos, que desempenham um papel fundamental durante a invasão do merozoíto de Plasmodium falciparum. Método. Revisão descritiva das características moleculares e estruturais dos receptores da superfície dos glóbulos vermelhos, que desempenham um papel fundamental durante a invasão do merozoíto de Plasmodium falciparum. Esta revisão foi baseada na pesquisa de literatura publicada até 2019 nas bases de dados eletrônicas especializadas na divulgação de pesquisas biomédicas. Foram encontrados 127 documentos, dos quais 111 foram selecionados e 33 foram excluídos por não apresentarem os critérios de inclusão, analisando um total de 78 referências. Conclusão. Nesta revisão, foram resumidas as características moleculares e estruturais dos receptores presentes nos glóbulos vermelhos, importantes no processo de invasão do merozoíto de P. falciparum. Também foi destacada a importância de elucidar as diferentes vias de invasão do parasita, a fim de desenvolver alternativas profiláticas ou terapêuticas que levem a mitigar ou eliminar a malária.


Subject(s)
Plasmodium falciparum , Erythrocytes , Merozoites , Malaria , Membrane Proteins
4.
Laboratory Medicine Online ; : 119-124, 2018.
Article in English | WPRIM | ID: wpr-715907

ABSTRACT

Pathogenic variants of bone morphogenic protein receptor type 2 gene (BMPR2) are related to the majority of cases of heritable pulmonary arterial hypertension (PAH). Over 400 pathogenic variants have been identified. However, clinical characterization of PAH is still incomplete. We present a case of heritable PAH in a Korean family showing serious clinical presentation with high penetrance. Genetic sequencing revealed a known heterozygous BMPR2 pathogenic variant, c.418+5G>A, at a splice site of intron 3. Serious clinical presentation with high penetrance suggested that the interplay of other factors with pathologic variants might be in genotype-phenotype correlation. Further studies are needed to clarify these issues for the development of personalized medicine approaches for PAH.


Subject(s)
Humans , Familial Primary Pulmonary Hypertension , Genetic Association Studies , Hypertension , Hypertension, Pulmonary , Introns , Penetrance , Precision Medicine , Pulmonary Artery
5.
Chinese Circulation Journal ; (12): 481-484, 2018.
Article in Chinese | WPRIM | ID: wpr-703885

ABSTRACT

Objectives: To investigate the genetic mutation in a Chinese family with Pulmonary Arterial Hypertension (PAH). Methods: Whole exome sequencing was performed in two patients and two healthy family members in the PAH pedigree. Patient-specific variations were screened by bioinformatics methods and compared between groups. To further identify the association between these variations and PAH, Sanger sequencing was used to analyze the genotype of PAH patients and 100 healthy controls. Results: Two affected persons were found among the eight family members. The patients was presented as dyspnea after exercise, and right-heart catheterization was performed to measure the mean pulmonary arterial pressure (mPAP, 77 mmHg), cardiac output (CO, 4.92 L/min), and pulmonary vascular resistance (PVR, 13.4 Wood units). The hereditary characteristic in this family presented in mother and child, suggesting an autosomal dominant patter. Exome sequencing, mutation detection and sanger variants validation revealed a novel heterozygous frameshift mutation (c.747_748 insCCTTTGATGGAACATGA:p.V250fs) in the BMPR2 gene. Meanwhile, this heterozygous insertion mutation was absent in 100 ethnically matched control samples screened by direct sanger sequencing. Conclusions: Our study revealed a novel heterozygous frameshift mutation in the BMPR2 gene, expanding the BMPR2 mutation spectrums.

6.
Chinese Critical Care Medicine ; (12): 156-161, 2017.
Article in Chinese | WPRIM | ID: wpr-510329

ABSTRACT

Objective To investigate the suppressive effect of carbon monoxide-releasing molecule Ⅱ (CORM-2) on LPS induced platelet α-granule exocytosis in sepsis via soluble N-ethylmaleimide-sensitive factor attached protein receptor/mammalian uncoordinated 18b (SNARE/Munc18b) complex formation.Methods Blood was collected from healthy volunteers' cubital vein, then platelets were isolated by differential centrifugation. Platelets were randomly divided into 5 groups. The control group did not undergo any treatment, the LPS group received 10 mg/L LPS simulation, the CORM-2 group and iCORM-2 group underwent LPS simulation and immediate administration of CORM-2 (10μmol/L and 50μmol/L) or iCORM-2 (50μmol/L), respectively. Samples were incubated in a CO2-incubator at 37 ℃, 95% humidity, and 5% CO2. Platelet α-granule contents were detected by using standard enzyme linked immunosorbent assay (ELISA), including platelet factor 4 (PF4), platelet derived growth factor-BB (PDGF-BB), and matrix metalloproteinase-2 (MMP-2). The expression of P-selectin was detected by flow cytometer. Transmission electron microscope and immunofluorescence microscope was used to assess platelet α-granules distribution. Expressions of Munc18b and SNARE proteins including vesicle-associated membrane protein-8 (VAMP-8), synaptosomal-associated protein-23 (SNAP-23) and syntaxin-11 (STX-11) were detected by Western Bolt. The SNARE/Munc18b complex formation was detected by immunoprecipitation.Results Compared with the control group, levels of PF4, PDGF-BB, MMP-2 and P-selectinin LPS-induced platelets were found to markedly elevated, while CORM-2 (10μmol/L and 50μmol/L) could decrease platelet α-granule contents exocytosis: [PF4 (μg/L): 7.69±0.58, 6.03±0.71 vs. 10.13±0.82; PDGF-BB (μg/L): 112.71±1.79, 102.91±5.86 vs. 128.78±1.39; MMP-2 (ng/L): 32.94±2.73, 27.58±3.36 vs. 53.26±1.21; P-selectin: (17.14±0.57)%, (15.35±0.68)% vs. (23.78±0.62)%; allP < 0.01]. Transmission electron microscope and immunofluorescence microscope showed that the extent of platelet α-granules assembled to platelet plasma membrane was significantly decreased following CORM-2 treatment. Compared with the control group, the expressions of Munc18b and SNARE proteins and SNARE/Munc18b complex formation in LPS-stimulated platelets were significantly increased, while CORM-2 (10μmol/L and 50μmol/L) inhibited these elevations (Munc18b/GAPDH: 0.80±0.08, 0.69±0.01 vs. 0.99±0.09; VAMP-8/GAPDH: 0.72±0.09, 0.50±0.12 vs. 1.18±0.14; SNAP-23/GAPDH: 1.18±0.22, 0.63±0.10 vs. 1.90±0.08; STX-11/GAPDH: 0.76±0.02, 0.57±0.08 vs. 1.16±0.23; VAMP-8/ Munc18b: 0.65±0.09, 0.53±0.07 vs. 1.21±0.20; SNAP-23/Munc18b: 0.85±0.07, 0.55±0.09 vs. 1.26±0.08; STX-11/ Munc18b: 0.78±0.05, 0.61±0.10 vs. 1.39±0.16; allP < 0.01). Above all, the data showed a dose dependent change.Conclusion We could suggest that CORM-2 suppressed α-granule exocytosis in LPS-stimulated platelets and the potential mechanisms might involve SNARE/Munc18b complex formation.

7.
Chinese Journal of Pharmacology and Toxicology ; (6): 836-841, 2015.
Article in Chinese | WPRIM | ID: wpr-477321

ABSTRACT

OBJECTIVE To investigate the effect and related mechanism of all ̄trans retinoic acid (atRA) exposure on osteogenic differentiation of mouse embryonic palate masenchymal cells MEPM. METHODS MEPM were cultured in osteogenic medium (OM) with atRA 0.1 and 1.0 μmol??L-1 for 1, 3,5, 7 and 9 d. MTT assay was performed to measure the cell viability. The alkaline phosphatase (ALP) activity was measured by chemical colorimetry. The cells were stained using the Von ̄Kossa technique to detect the formation of mineralization nodules after 21 d of culture. RT ̄PCR was performed to determine expression Runx2, osteopontin, bone morphogenetic protein receptor ( Bmpr) 1b, Bmpr2 and Smad5 mRNA. RESULTS The result of MTT on 9 d showed that, compared with normal control group, the cell viability of OM, OM+atRA 0.1 and 1.0 μmol??L-1 groups decreased significantly(P<0.01). Compared with normal control group, ALP activity of OM group increased significantly(P<0.05), while the ALP activity of OM+atRA 0.1 and 1.0 μmol??L-1 groups was lower than OM group(P<0.05). On 21 d, the Von ̄Kossa stai ̄ning results showed that the percentage of mineralization nodules formation of OM+atRA 1.0 μmol??L-1 group was (3.65±1.24)%, which was significantly lower than that of OM group(10.33±2.29)%(P<0. 05). On 9 d, the relative Run expression of OM group was the highest one in the four groups, while at ̄RA 1.0 μmol??L-1 treatment negatively regulated 20% in comparsion with OM group(P<0.05). Compared with normal control group, the mRNA expression of osteopontin of OM, OM+atRA 0.1 and 1.0 μmol??L-1 groups increased significantly(P<0.05); BDNF mRNA expression of OM group was 2.6 ̄fold to normal control group, while that of OM+atRA 1.0 μmol??L-1 group was 33% to OM group(P<0.05) . The level of Smad5 mRNA of OM+atRA 1.0 μmol??L-1 group was significantly lower than that of OM group(P<0.05). CONCLUSION atRA Might inhibit osteogenic differentiation of MEPM by down ̄regulated the expression of Bmpr1b.

8.
Journal of Forensic Medicine ; (6): 413-415,418, 2014.
Article in Chinese | WPRIM | ID: wpr-605043

ABSTRACT

Objective To investigate the expression of m onocyte chem otactic protein-1 (MCP-1) and its receptor CCchem okine receptor-2 (CCR-2) in coronary atherosclerosis plaques betw een sudden coro-nary death (SCD ) and non-SCD. Methods The expression levels of MCP-1 and CCR-2 in SCD group, coronary atherosclerosis group (non-SCD), control group (norm al coronary artery) w ere detected by im-m unohistochem istry. Results Positive rates of MCP-1 am ong the three groups w ere 78%, 47%, and 0%, respectively, w ith significant expressing differences betw een each tw o groups (P0.05). Conclusion O verexpression of MCP-1 and CCR-2 in coronary atherosclerotic plaques is closely correlated w ith SCD .

9.
Univ. sci ; 17(2): 119-139, may.-ago. 2012. tab
Article in Spanish | LILACS | ID: lil-669339

ABSTRACT

Objetivo. Construir un modelo molecular teórico de la estructura terciara del receptor 5HT2A de Homo sapiens a partir de estructurasobtenidas experimentalmente como plantillas. Materiales y métodos. Para la realización del modelo teórico se contempló el protocoloestablecido por Ballesteros y Weinstein para la construcción del receptor acoplado proteína G, por medio de alineamiento de la secuenciade aminoácidos, perfiles de hidrofobicidad, refinamiento de bucles por restricciones espaciales, y minimización de energía con el campode fuerza OPLS_2005. Resultados. El modelo obtenido fue validado por el gráfico de Ramachandran con un 91,7% de aminoácidosdentro de los límites establecidos para ángulos phi y psi, y un RMSD de 0,95 Å con respecto a rodopsina de bovino. Conclusiones. Seobtuvo un modelo teórico validado, útil para realización de estudios de acoplamiento ligando-receptor...


Objective Build a theoretical molecularmodel of the tertiary structure of the Homo sapiens 5HT2A receptor from experimentally obtained structures as templates. Materialsand methods In the construction of the theoretical model we considered the protocol established by Ballesteros and Weinstein for theconstruction of the G-protein coupled receptor, by the alignment of the amino acid sequence, hydrophobicity profiles, refinement ofloops by spatial restrictions and energy minimization with the force field OPLS_2005. Results The resulting model was validated bythe Ramachandran plot with 91.7% of amino acids within the limits set for angles phi and psi and a RMSD of 0.95 Å with respect tobovine rhodopsin. Conclusions We obtained a validated theoretical model useful in studies of ligand-receptor docking...


Objetivo. Construir um modelo molecularteórico da estrutura terciária do receptor 5HT2A de Homo Sapiens, com estruturas obtidas experimentalmente como moldes. Materiaise métodos. Para a elaboração do modelo teórico se utilizou o protocolo estabelecido por Ballesteros e Weinstein para a construção doreceptor acoplado à proteína G, por intermédio de alinhamento da seqüência de aminoácidos, perfis de hidrofobicidade, refinamento debucles por restrições espaciais e minimização da energia com o campo de força OPLS_2005. Resultados. O modelo obtido foi validadopelo gráfico de Ramachandran com 91,7% dos aminoácidos dentro dos limites estabelecidos para os ângulos phi e psi, e um RMSD de0,95 Å com respeito à rodopsina de bovino. Conclusões. Obteve-se um modelo teórico validado, útil para a realização de estudos deacoplamento ligante-receptor...


Subject(s)
Molecular Structure , Models, Molecular , GTP-Binding Proteins/analysis , GTP-Binding Proteins/classification , GTP-Binding Proteins/history
10.
Korean Journal of Medicine ; : 494-497, 2012.
Article in Korean | WPRIM | ID: wpr-12481

ABSTRACT

Pulmonary arterial hypertension is a rare life-threatening disorder. Inherited cases of this disorder are known as heritable pulmonary arterial hypertension. Familial cases of heritable pulmonary arterial hypertension are caused by germline mutations of the bone morphogenetic protein receptor type 2 gene (BMPR), a type II receptor of the TGF-beta superfamily. This has not been reported in Korea. We report the first case of familial hereditable pulmonary arterial hypertension in Korea. The family has a mutation in exon12 of the BMPR2 gene (c.2695C > T, p.R899X).


Subject(s)
Humans , Bone Morphogenetic Proteins , Germ-Line Mutation , Hypertension , Hypertension, Pulmonary , Korea , Transforming Growth Factor beta
11.
Korean Journal of Medicine ; : 238-242, 2012.
Article in Korean | WPRIM | ID: wpr-96836

ABSTRACT

Here, we describe the case of a 43-year-old male who was diagnosed with idiopathic pulmonary arterial hypertension and a mutation in the gene encoding bone morphogenetic protein receptor type 2 (BMPR2). The subject presented with hemoptysis and dyspnea on exertion and was diagnosed with pulmonary arterial hypertension. Genetic analysis revealed a novel deletion (c.1042_1047delGTTATT) in exon 8 of BMPR2. To the best of our knowledge, this is the first reported case of a BMPR2 mutation in a Korean patient with pulmonary arterial hypertension.


Subject(s)
Adult , Humans , Male , Bone Morphogenetic Proteins , Dyspnea , Exons , Hemoptysis , Hypertension , Hypertension, Pulmonary
12.
Yonsei Medical Journal ; : 740-745, 2010.
Article in English | WPRIM | ID: wpr-53350

ABSTRACT

PURPOSE: Several signaling pathways have been shown to regulate the lineage commitment and terminal differentiation of bone marrow stromal cells (BMSCs). Bone morphogenetic protein (BMP) signaling has important effects on the process of skeletogenesis. In the present study, we tested the role of bone morphogenetic protein receptor (BMPR) in the osteogenic differentiation of rat bone marrow stromal cells in osteogenic medium (OM) with or without BMP-2. MATERIALS AND METHODS: BMSCs were harvested from rats and cultured in OM containing dexamethasone, beta-glycerophosphate, and ascorbic acid, with or without BMP-2 in order to induce osteogenic differentiation. The alkaline phosphatase (ALP) activity assay and von kossa staining were used to assess the osteogenic differentiation of the BMSCs. BMPR mRNA expression was assessed using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The BMSCs that underwent osteogenic differentiation in OM showed a higher level of ALP activity and matrix mineralization. BMP-2 alone induced a low level of ALP activity and matrix mineralization in BMSCs, but enhanced the osteogenic differentiation of BMSCs when combined with OM. The OM significantly induced the expression of type IA receptor of BMPR (BMPRIA) and type II receptor of BMPR (BMPRII) in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively. CONCLUSION: BMSCs commit to osteoblastic differentiation in OM, which is enhanced by BMP-2. In addition, BMP signaling through BMPRIA and BMPRII regulates the osteogenic differentiation of rat BMSCs in OM with or without BMP-2.


Subject(s)
Animals , Male , Rats , Alkaline Phosphatase/metabolism , Bone Marrow Cells/cytology , Bone Morphogenetic Protein 2/pharmacology , Bone Morphogenetic Protein Receptors/genetics , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Culture Media/pharmacology , Osteogenesis/drug effects , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Stromal Cells/cytology
13.
Journal of Third Military Medical University ; (24)2003.
Article in Chinese | WPRIM | ID: wpr-555213

ABSTRACT

Objective To investigate the effects of 17?-estradiol (E2) on the gene expression of typeⅠA bone morphogenetic protein receptor (BMPR-ⅠA) and core-binding factor alpha 1 (Cbf?1) in rat bone marrow stromal cells exposed to the differentiation medium and to elucidate the effects of E2 on osteoblastogenesis. Methods Adherent bone marrow stromal cells were cultured in differentiation medium containing DEX (10 -7 mol/L), 1,25-(OH)2D3 (10 -9 mol/L) and different concentrations of E2. Effects of different concentrations of E2 on the gene expression of BMPR-ⅠA and Cbf?1 was quantified by RT-PCR based on the comparison with an internal reference, ?-actin expression, and identified by Northern blotting. Alkaline phosphatase (ALP) activity of cells was detected. Contents of type Ⅰ collagen were determined by Van Gieson staining. Results E2 could evidently inhibit the expression of BMPR-ⅠA and Cbf?1 mRNA during the differentiation process of bone marrow stromal cells into osteoblasts in a dose-dependent manner. These were confirmed by Northern blotting. The ALP activity increased in a concentration-dependent manner, but the amount of type Ⅰ collagen decreased in a concentration-dependent manner. Conclusion E2 can significantly inhibit the gene expression of BMPR-ⅠA and Cbf?1 in bone marrow stromal cells and inhibit osteoblastogenesis in vitro.

14.
Chinese Journal of Physical Medicine and Rehabilitation ; (12)2003.
Article in Chinese | WPRIM | ID: wpr-683335

ABSTRACT

Group A,the differences among the three groups were statistically significant(P0.05). TCR,CCR,DAP,OD,BRC in Groups B and C showed trends of increasing,the differences in terms of the contents of BMP-RⅠamong the 3 phases were statistically significant(P0.05);the ER in Group B and C showed a trend of decreasing,thee difference between 4- and 8-week and 4- and 12-week were signifieantly dif- ferent(P0.05).Conclusion After application of glucocorticoid for a short term,pathological changes maintained and showed trends of inereasing in early stage.HBO can reverse these changes.The outcome of 3-course HBO therapy is better than that of 1-course therapy.

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