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Canola (Brassica napus) is a widely cultivated species and provides important resources of edible vegetable oil, biodiesel production and animal feed. Seed development in Arabidopsis and canola shares a similar path: an early proliferation of endosperm to form a large seed cavity, followed by a second phase in which the embryo grows to replace the endosperm. In Arabidopsis, the seed reaches almost its final volume before the enlargement of the embryo. SHORT HYPOCOTYL UNDER BLUE1 (SHB1) is a key regulatory gene of seed development with a broad expression beyond endosperm development. By contrast, its two target genes, MINISEED3 (MINI3) and HAIKU2 (IKU2), are narrowly expressed in early developing endosperm and early embryo. We overexpressed SHB1 in canola to explore the possibility of altering seed development. As an alternative strategy, we expressed the canola IKU2 ortholog in Arabidopsis endosperm under the control of a stronger MINI3 promoter. SHB1 targeted canola orthologs of Arabidopsis MINI3 and IKU2 and caused a significantly increased seed mass. Overaccumulation of IKU2 in the early stage of Arabidopsis seed development also significantly increased the final seed mass. Our studies provide a strong case for increasing the final seed mass by manipulating endosperm proliferation at a rather early developmental stage in crops.
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Proteínas de Arabidopsis/genética , Arabidopsis/genética , Brassica napus/genética , Proteínas Quinases/genética , Sementes/crescimento & desenvolvimento , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/metabolismo , Brassica napus/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Mutação , Fenótipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Regiões Promotoras Genéticas , Proteínas Quinases/metabolismo , Sementes/química , Sementes/genética , Fatores de Transcrição/genéticaRESUMO
italic>Panax quinquefolium L. is a valuable Chinese herbal medicine with a large market demand. It has a complex chemical composition and numerous biological activities. At present, research on P. quinquefolium is focused on its underground parts, with especial interest in its saponins. There are few studies on non-saponins and the aboveground parts of Panax quinquefolium. Current quality standards are based on the saponin contents, which does not address the other benefits of Panax quinquefolium. This paper summarizes progress on the chemical components, pharmacological effects, quality evaluation, and product development of P. quinquefolium in recent years, and provides references for its further R&D and comprehensive utilization.
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Objective::To investigate the correlation between the characteristics and internal quality of Ophiopogonis Radix from Sichuan province through various quality evaluation, and provide reference for the formulation of grading standard of this herb. Method::For 28 batches of Ophiopogonis Radix, the characters, microscopic characteristics, thin-layer chromatography (TLC), the content of moisture, ash content, acid insoluble ash content, residues of sulfur dioxide, heavy metals, hazardous elements and paclobutrazol, water-soluble extract, contents of total saponins and three major components [ophiopogonin D (OPD), methylophiopogonanone A (MPOA) and methylophiopogonanone B (MPOB)] were determined.The relationships between various indicators and the grade of Ophiopogonis Radix were analyzed. Result::Ophiopogonis Radix exhibited specific properties in characters, microscopic characteristics and TLC.The contents of impurity, moisture, total ash, acid-insoluble ash and water-soluble extract existed differences in four grades of Ophiopogonis Radix from Sichuan province.For grade Ⅰ, grain number was 80-120 grains per 50 g, the moisture content was 11.1%-14.9%, total ash content was 1.6%-2.1%, acid-insoluble ash content was 0.03%-0.14%, water-soluble extract content was 77.0%-86.5%.For grade Ⅱ, grain number was 120-160 grains per 50 g, the moisture content was 13.1%-14.2%, total ash content was 1.3%-2.2%, acid-insoluble ash content was 0.06%-0.22%, water-soluble extract content was 75.9%-83.3%.For grade Ⅲ, grain number was 160-300 grains per 50 g, the impurity was 0.2%-8.4%, the moisture content was 12.6%-14.0%, total ash content was 1.2%-1.5%, acid-insoluble ash content was 0.06%-0.22%, water-soluble extract content was 74.0%-86.5%.For grade Ⅳ, grain number was 80-300 grains per 50 g, impurity was 1.2%-22.6%, the moisture content was 13.0%-15.4%, the total ash content was 1.4%-2.0%, acid-insoluble ash content was 0.03%-0.15%, water-soluble extract content was 79.8%-85.2%. Conclusion::It is reasonable and feasible to classify the grade of Ophiopogonis Radix from Sichuan province according to the appearance characteristics such as the grain number per 50 g and internal indexes such as impurity and water-soluble extract, which can be used as a reference for grading standards of Ophiopogonis Radix from Sichuan province.
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BACKGROUND@#Cerebrospinal fluid (CSF) has been demonstrated as a better source of circulating tumor DNA (ctDNA) than plasma for brain tumors. However, it is unclear whether whole exome sequencing (WES) is qualified for detection of ctDNA in CSF. The aim of this study was to determine if assessment of ctDNA in CSF by WES is a feasible approach to detect genomic alterations of glioblastoma.@*METHODS@#CSFs of ten glioblastoma patients were collected pre-operatively at the Department of Neurosurgery, Sun Yat-sen University Cancer Center. ctDNA in CSF and genome DNA in the resected tumor were extracted and subjected to WES. The identified glioblastoma-associated mutations from ctDNA in CSF and genome DNA in the resected tumor were compared.@*RESULTS@#Due to the ctDNA in CSF was unqualified for exome sequencing for one patient, nine patients were included into the final analysis. More glioblastoma-associated mutations tended to be detected in CSF compared with the corresponding tumor tissue samples (3.56 ± 0.75 vs. 2.22 ± 0.32, P = 0.097), while the statistical significance was limited by the small sample size. The average mutation frequencies were similar in CSF and tumor tissue samples (74.1% ± 6.0% vs. 73.8% ± 6.0%, P = 0.924). The R132H mutation of isocitrate dehydrogenase 1 and the G34V mutation of H3 histone, family 3A (H3F3A) which had been reported in the pathological diagnoses were also detected from ctDNA in CSF by WES. Patients who received temozolomide chemotherapy previously or those whose tumor involved subventricular zone tended to harbor more mutations in their CSF.@*CONCLUSION@#Assessment of ctDNA in CSF by WES is a feasible approach to detect genomic alterations of glioblastoma, which may provide useful information for the decision of treatment strategy.
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An ultra-performance liquid chromatography hybrid triple quadrupole-linear ion trap mass spectrometry(UPLC-QtrapMS) method was established to identify the metabolites in rat plasma,bile,urine and feces after oral administration of Cinnamomi Cortex(CC) aqueous extract. Several survey experiments,such as enhanced mass spectrum scan(EMS),precursor ion scan(PI),neutral loss scan(NL) and multiple ions monitoring(MIM) were applied to search target components,and two separate enhanced product ion(EPI) scans were triggered via information-dependent acquisition(IDA) method to generate the MS/MS spectra. According to the mass spectrometric data collected from reference standards and reported literature,the structures of metabolites were deduced. A total of76 metabolites and 5 original compounds were tentatively identified in rats after oral administration of CC aqueous extract. Deglycosylation,methylation,sulfonation,and glucuronidation were observed as the primary metabolic pathways for the chemical constituents of CC. These data are able to benefit the clarification of the therapeutic material basis,the clinical usage and further R&D of CC.
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Animais , Ratos , Administração Oral , Bile , Cromatografia Líquida de Alta Pressão , Cinnamomum zeylanicum , Medicamentos de Ervas Chinesas/metabolismo , Fezes , Espectrometria de Massas em TandemRESUMO
Objective: To discuss the phenotypic character and the HPLC fingerprints of radial striations from different germplasms Rehmanniae Radix. Method: The changes in the shape and column diameter of the radial striations of Rehmanniae Radix were observed and measured in the whole growth period. Besides,the HPLC fingerprints of the root,radial and un-radial striations were established to sign the chemical quality and analyzed by principal component analysis(PCA)and systematic cluster analysis. Result: There were significantly differences and regularities in the shape and proportion of the radial striations of different germplasms Rehmanniae Radix. The fingerprints showed the consistency between different types of chemical ingredients,and the differences in chemical quality characteristics mainly lay in the content of chemical compositions and theirs relative ratio. The results of PCA indicated that active ingredients, such as acteoside,catalpol,rehmaionoside D,rehmaionoside A and leonuride, were involved in the quality expression of different parts from various germplasms of Rehmanniae Radix,but each ingredient had a distinctive contribution rate to the differential quality expression between different parts from various germplasms of Rehmanniae Radix. However,the other components involved in the differential quality expression had different contribution rates in different germplasms.The systematic cluster analysis indicated that great differences in the chemical quality between the radial striations and un-radial striations of Beijing-1,Qinhuai,Qinhuai Zheng and 1706 germplasms,but with small differences in 85-5 and Baixuan germplasms. Conclusion: There are differences in phenotypic character of the radial striations and HPLC fingerprints between different germplasms Rehmanniae Radix.
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Atherosclerosis is an inflammatory disease of arterial wall caused by many factors, which is the main pathological basis of many cardiovascular diseases. Currently, "inflammatory response" theory is widely accepted as pathogenesis of atherosclerosis. Abnormal increase of apolipoprotein ApoB-100, a composition of low density lipoprotein (LDL), which causes pathological inflammation, is a major factor leading to atherosclerosis. Therefore, inhibition of ApoB-100 induced pathological inflammatory response by immunotherapy is expected to delay the development of atherosclerosis. This review focused on the recent advances of ApoB-100 vaccine and other ApoB-100 inhibitors against atherosclerosis.
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<p><b>OBJECTIVE</b>To study the effect of BAM bone grafting combined with inactivated autologous porous bone flap in repairing skull defect in rats.</p><p><b>METHODS</b>Seventy-two Wistar rats with skull defect were randomly divided into control group, inactivated autologous bone flap group (AB group), BAM bone-induced artificial bone material group (BAM group), and inactivated autologous bone flap with BAM bone-induced artificial bone group (BAM+AB group). The bone healing was evaluated with micro-CT and the new bone formation was assessed with histological staining at 1, 2, and 3 months after modeling.</p><p><b>RESULTS</b>Inactivated porous bone flap combined with BAM bone-induced artificial bone effectively induced vascular and fibrous tissue regeneration and osteogenesis in the cranial defects. With the inactivated porous bone flap as the scaffold, BAM bone-induced artificial bone obviously promoted the restoration of the skull appearance in the rats with cranial defects.</p><p><b>CONCLUSION</b>Inactivated autologous bone flap group and BAM bone-induced artificial bone material can promote skull healing and restoration of the original skull appearance, and can be used for reconstruction of the local anatomy of the skull surface.</p>
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The stomatal pores of plant leaves, situated in the epidermis and surrounded by a pair of guard cells, allow CO2 uptake for photosynthesis and water loss through transpiration. Blue light is one of the dominant environmental signals that control stomatal movements in leaves of plants in a natural environment. This blue light response is mediated by blue/UV A light-absorbing phototropins (phots) and cryptochromes (crys). Red/far-red light-absorbing phytochromes (phys) also play a role in the control of stomatal aperture. The signaling components that link the perception of light signals to the stomatal opening response are largely unknown. This review discusses a few newly discovered nuclear genes, their function with respect to the phot-, cry-, and phy-mediated signal transduction cascades, and possible involvement of circadian clock.
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Arabidopsis/fisiologia , Arabidopsis/efeitos da radiação , Luz , Estômatos de Plantas/fisiologia , Estômatos de Plantas/efeitos da radiação , Proteínas de Arabidopsis/metabolismo , Criptocromos/metabolismo , Modelos BiológicosRESUMO
Components that systematic separated from the root of Anaycclus pyrethrum were identified, in order to lay a foundation for future study of the root of A. pyrethrum. The CCK-8 assay showed that dichloromethane fraction exhibited the highest degree of cytotoxicity than others. Ten monomeric components were obtained from dichloromethane fraction and ethyl acetate fraction extracted from the root of A. pyrethrum, including 7 N-alkylamides, one coumarin and two flavonoid glycosides. They were identified as tetradeca-2E,4E,8E-trienoic acid 4-hydroxyphenylethylamide(1), deca-2E,4E-dienoicacid isobutylamide(2), undeca-2E,4E-diene-8,10-diynoic acid phenylethylamide(3), tetradeca-2E,4E-dienoic acid 4-hydroxyphenylethylamide(4), tetradeca-2E,4E-diene-8,10-diynoic acid isobutylamide(5), deca-2E,4E- dienoic acid 4-hydroxyphenylethylamide(6), dodeca-2E,4E-dienoic acid 4-hydroxy -phenyl-ethylamide(7), isoscopoletin(8), quercetin-7-O-β-D-glucopyranoside(9), isorhamnetin-7-O-β-D-glucopyranoside(10). Among them, compound 1 was identified as a new compound, Compounds 2-4, 8-10 were isolated from this herb for the first time.
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<p><b>OBJECTIVE</b>To investigate the correlation between the HLA genes and pathogenesis of aplastic anemia (AA), so as to find the susceptible AA genes.</p><p><b>METHODS</b>Polymerase chain reaction with specific sequence primers (PCR-SSP) method was used to detect the HLA typing of 50 AA patients and 183 normal healthy individuals as controls in Chinese Han population of northwestern plateau.</p><p><b>RESULTS</b>The frequency of HLA-A* 0201 (45.0%), B* 1501 (11.0%), B* 5501 (9.0%) and DRB1* 0901 (19.0%) gene frequences in AA patients were significantly higher than those in controls (Odds Ratio: OR=1.657, 2.138, 2.314 and 1.932, x2=4.882, 3.876, 3.863 and 4.473 (P<0.05). In contrast, A* 0301 gene frequency (4.0%) in AA was significantly lower than that in controls, OR=0.349, x2=4.154 (P<0.05). The male HLA-A* 0201 gene frequency was lower than that in female (38.2% vs 59.4%), and the difference was statistically significant (P<0.05). Concludsion: The HLA-A* 0201, B* 1501, B* 5501 and DRB1* 0901 genes may be considered as the risk markers while A* 0301 gene as a protective marker of AA, the HLA-A* 0201 also shows the sex differences.</p>
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Feminino , Humanos , Masculino , Alelos , Anemia Aplástica , Genética , Povo Asiático , Genética , China , Frequência do Gene , Cadeias HLA-DRB1 , Genética , Reação em Cadeia da Polimerase , Polimorfismo GenéticoRESUMO
Referring to the rapid developed life science and the higher requirements for the approval of innovative Chinese drugs in recent years, this paper described systematically the discovery, research and development (R&D) approaches for the innovative Chinese drugs under the new situation from the following five aspects, i. e., active components discovered from TCMs, the discovery of effective fractions of TCMs and their formulae, the R&D of TCM innovative drugs based on famous classic prescriptions and famous Chinese patent drugs, and the transformation of clinical effective prescriptions, on the basis of analysing the advantages of innovative drugs derived from natural products based on TCM theories and the problems existed in current R&D of new TCM drugs. Moreover, five suggestions are also given for the rapid development of TCM innovative drugs in China. All these will provide reference for the R&D of TCM innovative drugs.
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Humanos , China , Descoberta de Drogas , Medicamentos de Ervas Chinesas , Química , Farmacologia , Medicina Tradicional Chinesa , PesquisaRESUMO
Heterosis is a biological phenomenon whereby the offspring from two parents show improved and superior performance than either inbred parental lines. Hybrid rice is one of the most successful apotheoses in crops utilizing heterosis. Transcriptional profiling of F(1) super-hybrid rice Liangyou-2186 and its parents by serial analysis of gene expression (SAGE) revealed 1183 differentially expressed genes (DGs), among which DGs were found significantly enriched in pathways such as photosynthesis and carbon-fixation, and most of the key genes involved in the carbon-fixation pathway exhibited up-regulated expression in F(1) hybrid rice. Moreover, increased catabolic activity of corresponding enzymes and photosynthetic efficiency were also detected, which combined to indicate that carbon fixation is enhanced in F(1) hybrid, and might probably be associated with the yield vigor and heterosis in super-hybrid rice. By correlating DGs with yield-related quantitative trait loci (QTL), a potential relationship between differential gene expression and phenotypic changes was also found. In addition, a regulatory network involving circadian-rhythms and light signaling pathways was also found, as previously reported in Arabidopsis, which suggest that such a network might also be related with heterosis in hybrid rice. Altogether, the present study provides another view for understanding the molecular mechanism underlying heterosis in rice.
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Perfilação da Expressão Gênica , Vigor Híbrido/genética , Hibridização Genética/genética , Oryza/genética , Transcrição Gênica/genética , Ciclo do Carbono/genética , Redes Reguladoras de Genes/genética , Oryza/enzimologia , Oryza/metabolismo , Oryza/fisiologia , Fotossíntese/genética , Locos de Características Quantitativas/genéticaRESUMO
<p><b>OBJECTIVE</b>To investigate the effects of static magnetic fields (SMFs) with different exposure time on the maturation of rat osteoblasts in vitro and the expression of the estrogen receptor (ER) gene.</p><p><b>METHODS</b>The calvarial osteoblasts were isolated from newborn rats by enzyme digestion and randomly divided into 9 groups after one passage based on the exposure time of the SMFs[0 (control), 0.5 h, 1.0 h, 1.5 h, 2.0 h, 2.5 h, 3.0 h, 3.5 h, and 4.0 h]. The intensity was 3.9 mT in all SMFs. Those without SMFs exposure were used as the controls. The oeteoblasts were observed under the contrast phase microscope on a daily basis. After 48 h, cell proliferation was assayed by MTT method. The osteocalcin contents were measured after exposure to SMFs for 3 d, 6 d, 9 d, and 12 d. ERΑ and ERΒ mRNA expressions were measured by real-time PCR after SMFs treatment for 0 h, 24 h, 48 h, and 72 h.</p><p><b>RESULTS</b>Compared with the controls, the cell proliferation was significantly enhanced in the 2.0-h, 2.5-h, and 3.0-h groups (P<0.05). After SMFs treatment for 6 d, 9 d and 12 d, the 2.5-h group had significantly higher osteocalcin content than the control group did (P<0.05). After SMFs treatment for 0 h and 72 h, elevated ERΑ mRNA expression and reduced ERΒ mRNA expression were observed.</p><p><b>CONCLUSION</b>Exposure to SMFs, regardless of exposure time, is associated with enhanced cell proliferation, increased osteocalcin contents, and altered ERΑ and ERΒ mRNA expressions in opposite directions.</p>
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Animais , Ratos , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Campos Magnéticos , Osteoblastos , Biologia Celular , Metabolismo , Receptores de Estrogênio , Genética , MetabolismoRESUMO
This study is to investigate effects of genistein on rat femoral bone metabolic in vitro. Rat femoral tissues was isolated and randomly divided into two groups including control group and genistein (1 x 10(-5) mol x(-1)) group. Determinations of alkaline phosphatase (ALP) activity, calcium content and osteoprotegerin (OPG), type I-collagen (Collagen-I), RANKL, Runx-2 and bone morphogenetic protein (BMP-2) mRNA expression were done by real-time PCR. The results showed that 1 x 10(-5) mol x L(-1) genistein could increase the activity of ALP and contents of Ca, regulate bone metabolism activity of OPG, RANKL, BMP-2, Collagen-I and Runx-2 mRNA expression level. Genistein can significantly modulate bone metabolism related gene expression level of rat femoral tissue in vitro, and can increase calcium content and the activity of ALP.
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Animais , Ratos , Fosfatase Alcalina , Metabolismo , Proteína Morfogenética Óssea 2 , Genética , Metabolismo , Cálcio , Metabolismo , Colágeno Tipo I , Genética , Metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core , Genética , Metabolismo , Ativação Enzimática , Fêmur , Metabolismo , Regulação da Expressão Gênica , Genisteína , Farmacologia , Osteoprotegerina , Genética , Metabolismo , Fitoestrógenos , Farmacologia , Ligante RANK , Genética , Metabolismo , RNA Mensageiro , Metabolismo , Distribuição Aleatória , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo RealRESUMO
<p><b>OBJECTIVE</b>To investigate the effect of osthole on bone metabolism in rat femoral tissues in vitro.</p><p><b>METHODS</b>The rat femoral tissues were isolated in vitro. The optimal concentrations of ostehole (1×10(-5) mol/L) and estradiol (1×10(-8) mol/L) (the positive control) were selected by alkaline phosphatase activity (ALP). The ALP and calcium levels were detected by commmerical regents, and the expressions of osteoprotegerin, receptor activator of nuclear factor-κB ligand, runx-related gene 2, and bone morphogenetic protein-2 mRNA were determined by real-time reverse transcription-polymerase chain reaction.</p><p><b>RESULT</b>The osthole (1×10(-5) mol/L) significantly increased the activity of ALP, calcium level as well as the expressions of osteoprotegerin, receptor activator of nuclear factor-κB ligand, runx-related gene-2 and bone morphogenetic protein-2 mRNA in rat femoral tissues in vitro.</p><p><b>CONCLUSION</b>Osthole can improve calcium level and ALP activity and regulate the bone metabolism-related genes in rat femoral tissues.</p>
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Animais , Masculino , Ratos , Fosfatase Alcalina , Metabolismo , Proteína Morfogenética Óssea 2 , Metabolismo , Cálcio , Metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core , Metabolismo , Cumarínicos , Farmacologia , Fêmur , Metabolismo , Técnicas In Vitro , Osteoprotegerina , Metabolismo , Ligante RANK , Metabolismo , Ratos Sprague-DawleyRESUMO
<p><b>OBJECTIVE</b>To investigate the molecular mechanisms of icariin (ICA) in regulating the bone formation of osteoblasts and the bone resorption of osteoclasts.</p><p><b>METHODS</b>Primary osteoblast cell cultures were obtained from newborn rat calvarial. Calcified nodules were stained by alizarin red. The mRNA levels of osterix (OSX), runt-related transcription factor 2 (Runx-2), alkaline phosphatase (ALP), Collagen1, osteoprotegerin (OPG), and receptor activator of nuclear factor-ΚB ligand (RANKL) were analyzed by quantitative real-time RT-PCR, the protein levels of OPG, RANKL, and Collagen1 were examined by Western blotting, and the intracellular Ca(2+) concentration of osteoblasts was measured on a flow cytometer using the Cellquest program.</p><p><b>RESULTS</b>Compared with control group, ICA markedly promoted bone formation by significant up-regulating the gene expressions of OSX, Runx-2,ALP, and Collagen1, the protein expression of Collagen1(all P<0.01), and the Ca(2+) concentration. Furthermore, ICA remarkably inhibited bone resorption by significant up-regulating the mRNA and protein expressions of OPG as well as the OPG/RANKL ratio.</p><p><b>CONCLUSIONS</b>ICA could promote bone formation of osteoblasts through inducting the gene expressions of OSX,Runx-2, ALP and Collagen1, and the protein expressions of Collagen1, and by increasing the Ca (2+) concentration. Moreover, ICA could inhibit bone resorption of osteoclasts through regulating OPG/RANKL signal pathway.</p>
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Animais , Ratos , Fosfatase Alcalina , Metabolismo , Reabsorção Óssea , Células Cultivadas , Colágeno Tipo I , Metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core , Metabolismo , Flavonoides , Farmacologia , Expressão Gênica , Osteoblastos , Osteoclastos , Osteogênese , Osteoprotegerina , Metabolismo , Ligante RANK , Metabolismo , Ratos Sprague-Dawley , Receptor Ativador de Fator Nuclear kappa-B , Metabolismo , Fatores de Transcrição , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To compare the effect of icariin and genistein in the osteogenic differentiation of rat bone marrow stromal cells (rBMSC).</p><p><b>METHOD</b>Rat marrow stromal cells were seperated in vitro, and the optimal concentration of genisten and icriin were screened. Genistein and icariin with the concentration of 1 x 10(-5) mol x L(-1) were adopted to intereven rBMSCs cultured in vitro. Alkaline phosphatase (ALP) was determined at 3, 6, 9, 12,15 d after intervention; calcified nodule was detected with alizarin red staining at 12 d; OXS, Runx-2, bone morphogenetic protein (BMP-2) and Collagen-I mRNA expression were observed with Real-time RT-PCR at 12, 24, 48, 72, 96 h.</p><p><b>RESULT</b>Genistein and icariin with the concentration of 1 x 10(-5) mol x L(-1) could increase the activity of ALP and the content of Ca, regulate OXS, BMP-2, Runx-2 and Collagen-I mRNA expression.</p><p><b>CONCLUSION</b>Icariin showed a stronger effect in improving the osteogenic differentiation of rat bone marrow stromal cells than genistein.</p>
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Animais , Ratos , Fosfatase Alcalina , Genética , Metabolismo , Proteína Morfogenética Óssea 2 , Genética , Metabolismo , Diferenciação Celular , Células Cultivadas , Flavonoides , Farmacologia , Expressão Gênica , Genisteína , Farmacologia , Células-Tronco Mesenquimais , Biologia Celular , Metabolismo , Osteogênese , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta , Genética , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To investigate the effect of exposure to static magnetic fields (SMFs) of 3.9 mT on proliferation and differentiation of osteoblasts in vitro.</p><p><b>METHODS</b>The newborn rat calvarial osteoblasts were isolated by enzyme digestion and randomly divided into 9 groups after one passage. The intensity of the SMFs was 3.9 mT. The cells were exposed in the SMFs for 0 (control group), 0.5, 1.0, 1.5, 2.0, 2.5, 3, 3.5 and 4.0 h groups respectively. They were observed under the contrast phase microscope each day. After 48 h, cell proliferation was assayed by MTT method. The alkaline phosphatase (Alkaline Phosphatase, ALP) activities and calcium content were measured after 3, 6, 9, and 12 days exposed with SMFs. The ALP positive colonies were histochemically stained after 8 days and the calcified nodules were stained by Alizarin Bordeaux after 10 days; BMP-2, Runx-2 and Opg mRNA expression were measured after SMFs treatment in 0, 24, 48 and 72 h.</p><p><b>RESULTS</b>Contrast with control group, all SMFs groups enhanced cell proliferation (P < 0.01 or P < 0.05), and they promoted maturation and mineralization of the osteoblasts. The results showed that SMFs improved the ALP activity, promoted calcium content, boost BMP-2, Runx -2 and Opg mRNA expression.</p><p><b>CONCLUSION</b>The cells exposed to the SMFs of 3.9 mT at 2.5 h apparently promote proliferation and differentiation of osteoblasts in vitro.</p>
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Animais , Ratos , Proteína Morfogenética Óssea 2 , Genética , Cálcio , Metabolismo , Diferenciação Celular , Efeitos da Radiação , Proliferação de Células , Efeitos da Radiação , Subunidade alfa 1 de Fator de Ligação ao Core , Genética , Campos Magnéticos , Osteoblastos , Fisiologia , Efeitos da Radiação , Osteoprotegerina , Genética , Ratos Sprague-Dawley , Fatores de TempoRESUMO
This study is to compare the effects of kaempferide and anhydroicaritin on biomineralization of rat osteoblasts (ROB) in vitro. Calvarias were dissected aseptically from newborn SD rats, the osteoblasts were obtained by enzyme digestion and were cultured in MEM containing 10% FBS. The medium was changed every three days, and serial subculture was performed when cells covered with 90% of the dish. Kaempferide and anhydroicaritin were separately added with final concentrations of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) and 1 x 10(-7) mol x L(-1) under the conditions of osteogenic differentiation. The proliferation was measured by MTT, and the optimal concentration was detected by the ALP activity at the 9th day after osteogenic induction culture. The osteogenic indexes of kaempferide, anhydroicaritin and control group with the optimal concentration were compared. The result showed that the anhydroicaritin at concentration of 1 x 10(-5) mol x L(-1) had significantly promoted the activity of ALP, calcium content and osteocalcin content, increased the number of CFU-F(ALP) and mineralized nodules, enhanced the mRNA level of BMP-2, OSX and Runx-2, which are key genes of osteogenic differentiation, and raised the protein content of collagen-I. However, the kaempferide group had not significantly represented the ability that promoted osteogenic differentiation of ROB. The difference of osteogenic differentiation on ROB between kaempferide and anhydroicaritin was caused by the prenyl group on C-8 of icariin.