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2.
Front Plant Sci ; 13: 1043761, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36438137

RESUMO

Red sage, the dry root and rhizome of the herbaceous plant Salvia miltiorrhiza Bunge, is widely used for treating various diseases. The low content of tanshinones (terpenoids) has always restricted development of the S. miltiorrhiza industry. Here, we found that SmDXS5, a rate-limiting enzyme-coding gene located at the intersection of primary and secondary metabolism, can effectively change the transcription level and secondary metabolome profile of hairy roots of S. miltiorrhiza, and significantly increase the content of tanshinones. Agrobacterium rhizogenes was used to infuse S. miltiorrhiza explants, and hairy roots of S. miltiorrhiza expressing the SmDXS5 gene were obtained successfully. We identified 39 differentially accumulated metabolites (DAMs) by metabolomics based on ultra-high performance liquid chromatography quadrupole exactive mass spectrometry and multivariate statistics. These DAMs might be key metabolites of SmDXS5 gene regulation. RNA sequencing was used to compare gene expression between the hairy roots of the SmDXS5 overexpressing group and the blank control (BC) group. Compared with the BC group, 18,646 differentially expressed genes were obtained: 8994 were upregulated and 9,652 downregulated. The combined transcriptome and metabolome analyses revealed that the mevalonate and methylerythritol phosphate pathways and synthase gene expression levels in the SmDXS5 overexpressing group were upregulated significantly, and the accumulation of tanshinone components was increased significantly, which promoted the process of glycolysis and promoted the transformation of carbohydrates to secondary metabolism. Moreover, the expression of SmPAL, the first rate-limiting enzyme gene of the phenylpropane pathway, decreased, reducing the accumulation of phenolic acid, another secondary metabolite. Therefore, SmDXS5 can be defined as a 'valve' gene, mainly responsible for regulating the distribution of primary and secondary metabolic flow of tanshinones in S. miltiorrhiza, and for other secondary metabolic pathways. The discovery of SmDXS5 and its molecular valve function in regulating primary and secondary metabolism will provide a basis for the industrial production of tanshinone components, and cultivation of high quality S. miltiorrhiza.

3.
Life Sci ; 242: 117240, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31891722

RESUMO

Lycium barbarum polysaccharides (LBP) are derived from Wolfberry and have antioxidant activities. This study aimed to evaluate the efficacy of LBP for kidney injury in a rat model of sepsis. Male rats were divided randomly to control group (Con), LPS group (LPS), ulinastatin group (ULI), low dose LBP group (LBP-1), middle dose LBP group (LBP-2) and high dose LBP group (LBP-3). After intraperitoneal injection of LPS (5 mg/kg) to make sepsis model (LPS group), 10,000 U/kg ulinastatin were given in ULI group, and 200, 400 and 800 mg/kg LBP was given in LBP-1, -2, -3 group, respectively. Serum IL-1ß, IL-6, IL-8, TNF-α and NF-κB levels were measured by ELISA. Nrf2, Keap1, NF-κB, HO-1 and NQO1 expression levels were detected by PCR and Western blot analysis. We found that LBP decreased the levels of NF-κB and pro-inflammatory cytokines while attenuated kidney injury. In addition, LBP regulated Keap1-Nrf2/ARE signaling pathway in the kidney. In conclusion, LBP attenuates inflammation injury in the kidney via possible regulation of Keap1-Nrf2/ARE signaling.


Assuntos
Injúria Renal Aguda/prevenção & controle , Elementos de Resposta Antioxidante/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Sepse/complicações , Transdução de Sinais/efeitos dos fármacos , Animais , Western Blotting , Citocinas/metabolismo , Modelos Animais de Doenças , Masculino , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real
4.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 37(4): 446-50, 2015 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-26564463

RESUMO

OBJECTIVE: To observe the morphology of hypertrophic scar tissue and explore the expressions and distribution of vascular endothelial growth factor (VEGF) and transforming growth factor beta activated kinase 1(TAK1 )in these tissues. METHOD: Hematoxylin-eosin staining, Masson staining,immunofluorescence,and real-time polymerase chain reaction were used to detect the localization and expression of VEGF and TAK1 in 15 hypertrophic scar tissues and 10 normal skin tissues. RESULTS: Morphological observation showed that the dermal fibroblasts in hypertrophic scar were disorderly and densely arranged (compared to the normal skin). Immunofluorescence displayed that the expressions of VEGF and TAK1 in hypertrophic scar tissue were higher than in normal skin tissues. Real-time polymerase chain reaction showed the mRNA expressions of both VEGF and TAK1 were significantly higher in hypertrophic scar tissue than in normal tissue (P<0.01, P<0.05,respectively). CONCLUSIONS: Hypertrophic scar tissue has higher collagen fibrosis degree and higher TAK1 and VEGF expressions than the normal skin. VEGF and TAK1 can be used as the reference indicators for the diagnosis and differential diagnosis of hypertrophic scar and serve as new therapeutic targets.


Assuntos
Cicatriz Hipertrófica , Forma Celular , Colágeno , Fibroblastos , Humanos , MAP Quinase Quinase Quinases , Fator de Crescimento Transformador beta , Fator A de Crescimento do Endotélio Vascular
5.
Molecules ; 19(10): 16925-36, 2014 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-25340298

RESUMO

As one of the most important components of Panax ginseng, ginsenoside Rg1 has certain anti-aging effects, improving the activity of learning and memory. Studies have showed that ginsenoside Rg1 improves the memory impairment associated with Alzheimer's disease (AD). In this study, the effects of ginsenoside Rg1 were investigated through the activity of toll-like receptor (TLR) 3, TLR4 and their signaling transduction pathways in amyloid ß peptide 25-35 (Aß25-35) induced AD cell model. Thus we investigated several critical components of the TLR pathway. The neuroglial cell line NG108-15 was stimulated with or without Aß25-35, while different concentrations of ginsenoside Rg1 were administered. After 24 h, tumor necrosis factor-α (TNF-α), interferon-ß (IFN-ß) in cell supernatant and inducible nitric oxide synthase (iNOS) in cell lysate supernatant were measured with enzyme-linked immunosorbent assays (ELISAs). The mRNA and protein expression of TLR3, TLR4, nuclear factor kappa B (NF-κB) and tumor necrosis factor receptor-associated factor-6 (TRAF-6) were detected by real-time PCR and western blot methods, respectively. The experimental results showed that Aß25-35 could markedly raise the level of TNF-α, IFN-ß and iNOS, and increase the expressions of mRNA and TLR3, TLR4, NF-κB and TRAF-6 protein in the NG108-15 cells. At the same time, the ginsenoside Rg1 significantly reduced the expressions of proteins and mRNA of TLR3, TLR4, NF-κB and TRAF-6, and down-regulated the levels of TNF-α, IFN-ß of cell supernatant and iNOS of cell lysate supernatant in a concentration-dependent manner. In conclusion, ginsenoside Rg1 has good activity for suppressing the signaling transduction pathway of TLR3 and TLR4, and decreasing the inflammation factors induced by Aß25-35 in NG108-15 cells, and this may be the mechanism of ginsenoside Rg1 action in AD treatment, but more studies are needed to identify its specificity.


Assuntos
Doença de Alzheimer/tratamento farmacológico , Regulação da Expressão Gênica/efeitos dos fármacos , Ginsenosídeos/farmacologia , Mediadores da Inflamação/metabolismo , Neuroglia/metabolismo , Receptor 3 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Doença de Alzheimer/induzido quimicamente , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/toxicidade , Animais , Western Blotting , Células Cultivadas , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Interferons/metabolismo , Camundongos , NF-kappa B/metabolismo , Neuroglia/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II/metabolismo , Panax/química , Fragmentos de Peptídeos/toxicidade , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Receptor 3 Toll-Like/genética , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa/metabolismo
6.
Zhongguo Zhong Yao Za Zhi ; 39(7): 1199-202, 2014 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-25011253

RESUMO

Tibetan Herbal medicine has its own complete theory based on five sources doctrine. And the theories of "Liuwei", "Baxing" and "Shiqi Gongxiao" formed the basic core components of the property theory of Tibetan medicine. However, books and literature of Tibetan medicine have never been systematically expounded and discussed about it specially which thus will limit the further development of Tibetan medicine theory. In this thesis, we firstly introduced three basic core components of the property theory-the "Liu Wei", "Baxing", and "Shiqi Gongxiao" and their interactions as well. At the same time, the links and similarities between the theory of Tibetan medicine and Chinese medicine theory were compared. The job of the thesis done above is to lay the foundation for further systematic reveal and development of Tibetan medicine theory.


Assuntos
Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Medicina Tradicional do Leste Asiático , Plantas Medicinais/química , Humanos , Fitoterapia
7.
Zhongguo Zhong Yao Za Zhi ; 38(8): 1134-7, 2013 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-23944024

RESUMO

OBJECTIVE: To develop an effective identification method for accurately discriminating Psammosilene tunicoides and its confused species by the combined method of microscopic identification and molecular identification, so-called systematic identification of Chinese materia medica (SICMM). METHOD: P. tunicoides and its confused species were accurately discriminated by SICMM method, which was established by comprehensively use of microscopic identification and DNA identification method. The DNA identification included the following analysis: the BLAST alignment, specific bases and N-J phylogenetic tree analysis. RESULT: The cluster crystals were not observed in P. tunicoides, but great deals of them were found in Silene viscidula. Further more, big differences of ITS sequence were observed and analyzed between P. tunicoides and its confused specie of S. viscidula. CONCLUSION: The system method is a scientific and accurate method for the identification of P. tunicoides and its counterfeit species.


Assuntos
Caryophyllaceae/classificação , Caryophyllaceae/genética , DNA Intergênico , Filogenia , Sequência de Bases , Caryophyllaceae/química , Caryophyllaceae/citologia , Fenótipo , Alinhamento de Sequência
8.
Zhongguo Zhong Yao Za Zhi ; 38(9): 1451-4, 2013 May.
Artigo em Chinês | MEDLINE | ID: mdl-23944087

RESUMO

This paper put forward a more accurate identification method for identification of Chinese materia medica (CMM), the systematic identification of Chinese materia medica (SICMM) , which might solve difficulties in CMM identification used the ordinary traditional ways. Concepts, mechanisms and methods of SICMM were systematically introduced and possibility was proved by experiments. The establishment of SICMM will solve problems in identification of Chinese materia medica not only in phenotypic characters like the mnorphous, microstructure, chemical constituents, but also further discovery evolution and classification of species, subspecies and population in medical plants. The establishment of SICMM will improve the development of identification of CMM and create a more extensive study space.


Assuntos
Medicamentos de Ervas Chinesas , Materia Medica
9.
Zhongguo Zhong Yao Za Zhi ; 37(24): 3777-83, 2012 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-23627178

RESUMO

OBJECTIVE: To analyse the polymorphism of squalene synthase gene and reveal the influence of squalene synthase (SQS) gene polymorphism on the catalytic efficiency of its encode enzyme in Glycyrrhiza uralensi. METHOD: The total RNA was extracted. PCR was used to amplify the coding sequences of squalene synthase gene, which were sequenced and analysed. The expression vectors containing different SQS gene sequences, including SQS1C, SQS1F, SQS2A, SQS2B, were constructed and transformed into Escherichia coli BL21. The fusion protein was induced to express by IPTG, then was isolated, purified and used to carry out the enzymatic reaction in vitro. GC-MS was used to analyse the production. RESULT: There were three kinds of gene polymorphism existing in SQS1 gene of G. uralensis, including single nucleotide polymorphism (SNPs), insertion/deletion length polymorphism (InDels) and level of amino acid, the proportion of conservative replace of SQS1 was 53.94%, and there were 2 mutational sites in structural domains. The proportion of conservative replace of SQS2 was 60%, and there was 1 mutational site in structural domains. The production squalene could be detected by GC-MS in all the 4 kinds of enzymatic reactions. The capacity of accumulating squalene of SQS1F was higher than other SQS genes. CONCLUSION: The polymorphism of SQS gene was quite abundant in G. uralensis, which maybe the molecular foundation of the formation of high-quality liquorice.


Assuntos
Farnesil-Difosfato Farnesiltransferase/genética , Glycyrrhiza uralensis/genética , Proteínas de Plantas/genética , Polimorfismo Genético , Substituição de Aminoácidos , Biocatálise , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Farnesil-Difosfato Farnesiltransferase/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Glycyrrhiza uralensis/enzimologia , Mutação INDEL , Isoenzimas/genética , Isoenzimas/metabolismo , Dados de Sequência Molecular , Proteínas de Plantas/metabolismo , Polimorfismo de Nucleotídeo Único , Proteínas Recombinantes/metabolismo , Análise de Sequência de DNA , Esqualeno/metabolismo
10.
Zhongguo Zhong Yao Za Zhi ; 37(24): 3784-8, 2012 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-23627179

RESUMO

OBJECTIVE: To analyse the effect of expression proteins containing different escherichia coli of 3-hydroxy-3-methylglutary-coenzyme A reductase(HMGR) genic mutation on the conversion efficiency of MVA with GC-MS method, in order to lay a foundation for revealing the function of HMGR gene polymorphism of Glycyrrhiza uralensis in the production of high-quality G. uralensis medicines. METHOD: The expression carrier was established from four HMGR genic mutation types cloned from G. uralensis and transformed into Escherichia coli BL21. The protein was induced to express, detected and purified. The purified protein was adopted for in vitro enzymatic reaction. TLC and GC-MS were used for qualitative and quantitative analysis on reaction products. RESULT: The catalytic activity of L/V genotype(-HSL and -HSV) was similar, and so was the catalytic activity of the genotype with GA insertion (GALLV and GALSV), but the catalytic activity of the latter was around 2 times higher than that of the former. CONCLUSION: The functional gene polymorphism of G. uralensis may be the molecular foundation for the production of high-quality G. uralensi medicines.


Assuntos
Glycyrrhiza uralensis/genética , Hidroximetilglutaril-CoA Redutases/genética , Proteínas de Plantas/genética , Polimorfismo Genético , Biocatálise , Cromatografia em Camada Fina , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Cromatografia Gasosa-Espectrometria de Massas , Glycyrrhiza uralensis/enzimologia , Hidroximetilglutaril-CoA Redutases/metabolismo , Mutação , Proteínas de Plantas/metabolismo , Proteínas Recombinantes/metabolismo
11.
Zhongguo Zhong Yao Za Zhi ; 37(24): 3789-92, 2012 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-23627180

RESUMO

OBJECTIVE: To reveal the 3-hydroxy-3-methylglutary-coenzyme A reductase (HMGR) gene polymorphism of Glycyrrhiza uralensis, and the correlation between HMGR gene polymorphism and the content of glycyrrhizic acid. METHOD: Liquorice plants containing different content of glycyrrhizic acid were used as materials. RT-PCR was used to amplify their HMGR gene sequences, which were connected with vector pMD19-T for clone sequencing. Multiple alignments were performed to analyse HMGR gene polymorphism of G. uralensis. Then the correlation between HMGR gene polymorphism and the content of glycyrrhizic acid was revealed. RESULT: HMGR gene sequences polymorphism included codon mutation, base substitution mutation, copy number polymorphism and allele heterozygosity. There were 4 types of mutations in HMGR gene coding amino acid sequences, namely -HSL, -HSV, GALLV, GALSV. Among them, -HSV type was common in liquorice plants, -HSL type only existed in liquorice plants with low content of glycyrrhizic acid, and GALSV type only existed in liquorice plants with high content of glycyrrhizic acid. CONCLUSION: HMGR gene sequences of G. uralensis are highly polymorphic and related to the content of glycyrrhizic acid.


Assuntos
Glycyrrhiza uralensis/genética , Ácido Glicirrízico/metabolismo , Hidroximetilglutaril-CoA Redutases/genética , Proteínas de Plantas/genética , Polimorfismo Genético , Clonagem Molecular , DNA Complementar/química , DNA Complementar/classificação , DNA Complementar/genética , Glycyrrhiza uralensis/enzimologia , Glycyrrhiza uralensis/metabolismo , Hidroximetilglutaril-CoA Redutases/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Mutação , Filogenia , Proteínas de Plantas/metabolismo , Análise de Sequência de DNA
12.
Yao Xue Xue Bao ; 45(6): 785-90, 2010 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-20939191

RESUMO

Acetyl-CoA C-acetyltransferase (AACT) is the first enzyme in the terpene synthesis pathway, catalyzed two units of acetyl-CoA to acetoacetyl-CoA. In order to study the tanshinone biosynthesis in Salvia miltiorrhiza, a novel AACT gene, SmAACT, was cloned using cDNA microarray and RACE strategy. The full length cDNA of SmAACT is 1 623 bp (accession No. EF635969), which contained a 1 200 bp open reading frame (ORF) encoding a 399 amino acid protein. Nine introns were found in the genomic sequence. SmAACT was upregulated by YE and Ag+ elicitors both with cDNA microarray and quantitative RT-PCR analyses along with the accumulation of tanshinones. Sequence homology comparison and phylogenetic analysis all suggested that SmAACT belonged to the class of acetyl-CoA C-acetyltransferase. The transcription level of SmAACT was relatively higher in root than that in stem and leaf tissues. SNP analysis revealed that SmAACT was highly variable in the region of 6 to 9 introns with 33 SNPs in the 600 bp region, there are 5 SNPs in the cDNA region while they are all synonymous cSNPs. Some special genotypes were found in Salvia miltiorrhiza from different areas. SmAACT will be an useful gene for further analyze the mechanism of gene regulation among the tanshinones biosynthesis.


Assuntos
Acetil-CoA C-Acetiltransferase/genética , Plantas Medicinais/enzimologia , Polimorfismo de Nucleotídeo Único , Salvia miltiorrhiza/enzimologia , Acetil-CoA C-Acetiltransferase/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Genótipo , Íntrons , Análise de Sequência com Séries de Oligonucleotídeos , Fases de Leitura Aberta , Filogenia , Folhas de Planta/enzimologia , Folhas de Planta/genética , Raízes de Plantas/enzimologia , Raízes de Plantas/genética , Caules de Planta/enzimologia , Caules de Planta/genética , Plantas Medicinais/classificação , Plantas Medicinais/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Salvia miltiorrhiza/classificação , Salvia miltiorrhiza/genética
13.
Yao Xue Xue Bao ; 43(12): 1251-7, 2008 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-19244759

RESUMO

This paper firstly introduced the acquired full length cDNA of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol kinase from hairy roots of Salvia miltiorrhiza (Abbr: SmCMK, GenBank number: EF534309). Results of KEGG analysis showed that SmCMK was belong to the upstream of nonemevalonate pathway, the only one kinase of the pathway. The full-length cDNA was deduced as encoding 4-(cytidine 5'-diphospho)-2-C-methylerythritol kinase (designated as SmCMK), and the sequence had a 1493 bp including 5' UTR 71 bp and 3' UTR 232 bp, an open reading frame (ORF) encoding a protein of 396 amino acid residues. The deduced protein had isoelectric point (pI) of 6.78 and a calculated molecular weight about 43 kDa, similar to cloned diterpene of CMK from other species of plants such as Mentha piperita and Lycopersicon esculentum reported previously. Real time PCR results indicated that elicitors of MJ stimulated the increase of mRNA expression of SmCMK. At the same time, results of high performance liquid chromatography (HPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy root of Salvia miltiorrhiza were increased dramatically after treated with methyl jasmonate (MJ). This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by MJ. It proved primarily that the increased expression level of mRNA of SmCMK helps to enhance tanshinones' accumulation, which will be the basis for further study on the mechanism of gene regulation of secondary metabolism of tanshinones.


Assuntos
Fosfotransferases (Aceptor do Grupo Álcool)/genética , Proteínas de Plantas/genética , Salvia miltiorrhiza/enzimologia , Abietanos , Acetatos/farmacologia , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Sequência Conservada , Ciclopentanos/farmacologia , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Dados de Sequência Molecular , Fases de Leitura Aberta , Oxilipinas/farmacologia , Fenantrenos/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/enzimologia , Plantas Medicinais/enzimologia , Plantas Medicinais/genética , Plantas Medicinais/metabolismo , RNA Mensageiro/metabolismo , Salvia miltiorrhiza/genética , Salvia miltiorrhiza/metabolismo , Homologia de Sequência de Aminoácidos
14.
Zhongguo Zhong Yao Za Zhi ; 32(14): 1393-5, 2007 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-17966348

RESUMO

OBJECTIVE: To study the metallothionein genes of Salvia miltiorrhiza through bioinformatics and characterization of tissue expression in regenerated shoots. METHOD: Metallothionein genes were obtained by cDNA microarray analyze. BLAST was used for align, ORF finder software was used to find open reading frame, Prosite database was used to analyze the protein. Semi-quantitative RT-PCR method was used to detect the gene expression level. RESULT: Two metallothionein genes were obtained which were contained a deduced amino acid sequence of 80 and 79 residues, named as SmMT-2a and SmMT-2b, they had a homology of 71.25%. Semiquantitative RT-PCR indicated that metallothionein genes were expressed in all tissues such as root, stem and leaf in regenerated shoots, while the expression level was higher in leaf than in root and stem. CONCLUSION: It was the first time that metallothionein genes were obtained from S. miltiorrhiza. It provides a good basis for further functional study of S. miltiorrhiza.


Assuntos
Genes de Plantas , Metalotioneína/genética , Plantas Medicinais/genética , Salvia miltiorrhiza/genética , Sequência de Aminoácidos , Sequência de Bases , DNA Complementar/genética , Regulação da Expressão Gênica de Plantas , Genômica , Metalotioneína/metabolismo , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Fases de Leitura Aberta , Plantas Medicinais/metabolismo , Salvia miltiorrhiza/metabolismo , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos
15.
Zhongguo Zhong Yao Za Zhi ; 32(12): 1137-41, 2007 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-17802870

RESUMO

OBJECTIVE: Establishing cDNA microarray, in order to study functional genomics of Salvia miltiorrhiza. METHOD: Total RNA samples were prepared from S. miltiorrhiza roots using a modified CTAB method. mRNA was isolated by Quichprep Micro mRNA Purification Kit from Pharmacia. Then cDNA was synthesized and cloned into the EcoRI-XhoI sites of the ZAP Express vector using a cDNA synthesis kit, and the ligation mixture was packaged using a ZAP-cDNA Gigapack Gold III cloning kit (Stratagene). The single phage was isolated for PCR amplification, Aliquots of the PCR reactions were analyzed in a 1.5% agarose gel to verify the quality of PCR. The remaining cDNA was purified by Multiscreen filter plates (Millipore) and aliquots were analyzed by agarose gel again to verify the quality of purification. Clones passed verification was resuspended in 15 microL 50% DMSO for arraying. An actin gene from S. miltiorrhiza was used for positive control. PloyA and 50% DMSO was used for negative controls. RESULT: Bacterial colonies containing cNDAs of S. miltiorrhiza were inserted with average insert size of 0. 5 kb to 2. 5 kb. Total 4 354 genes were singled out from the first 8 736 PCR product and used for cDNA microarray manufacture. Single color fluorescence hybridization showed that all positive controls had signals while negative controls had no signals. CONCLUSION: It was the first cDNA microarray about traditional Chinese herbs especially for geoherbs. It could be a powerful tool for studying functional genomics of S. miltiorrhiza.


Assuntos
Genômica/métodos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Plantas Medicinais/genética , Salvia miltiorrhiza/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Raízes de Plantas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa
16.
Zhongguo Zhong Yao Za Zhi ; 32(4): 300-2, 2007 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-17455461

RESUMO

OBJECTIVE: To study the effects of methyl jasmonate (MJ) on the accumulation and release of tanshinones in suspension cultures of Salvia miltiorrhiza hairy roots. METHOD: After 18 day's suspension culture of S. miltiorrhiza hairy roots induced by Agrobacterium rhizogenes ATCC15834, the chemical elicitor--methyl jasmonat was added into 6-7V suspension cultures and at the same time, tanshinones contents (including cryptotanshinone and tanshinone II(A)) on the day 2, 6 and 9, after dealing with MJ, was quantified by HPLC. RESULT: After dealing with MJ on the day 2, 6 and 9, the concentration of cryptotanshinone reached to 0.039, 0.204, 0.571 mg x g(-1) respectively,and tanshinone II(A) reached 0.251, 0.601 and 1.563 mg x g(-1) respectively. After 9 day's treatment by MJ, the maximum increase of cryptotanshinone and tanshinone II(A) were 23.8 fold and 6.2 fold higher than that of the control respectively. CONCLUSION: MJ could stimulate the accumulation of tanshinones in S. miltiorrhiza and have released them into the culture medium.


Assuntos
Acetatos/farmacologia , Ciclopentanos/farmacologia , Oxilipinas/farmacologia , Fenantrenos/metabolismo , Raízes de Plantas/efeitos dos fármacos , Salvia miltiorrhiza/efeitos dos fármacos , Abietanos , Técnicas de Cultura , Reguladores de Crescimento de Plantas/farmacologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Plantas Medicinais/efeitos dos fármacos , Plantas Medicinais/crescimento & desenvolvimento , Plantas Medicinais/metabolismo , Salvia miltiorrhiza/crescimento & desenvolvimento , Salvia miltiorrhiza/metabolismo
18.
J Vasc Surg ; 43(6): 1090-5, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16765220

RESUMO

OBJECTIVE: To evaluate early and mid-term results of endovascular repair of acute type B aortic dissection by stent graft. METHODS: From June 2001 to May 2005, 63 patients with acute type B aortic dissection underwent stent graft implantation. The study included 59 men and 4 women with an average age of 50.4 +/- 11.4 years (range, 31-80 years). Four patients underwent stent-graft implantation in the acute phase. Fifty-nine patients with acute type B dissection underwent stent-graft implantation 2 weeks after the onset of dissection. All patients were followed up from 1 to 47 months (average, 11.7 +/- 10.6 months). The clinical data of the patients were analyzed. RESULTS: The primary tear was incompletely sealed in three cases. The incidence of incomplete seal was 4.8%. Ascending aortic dissection occurred in three cases. One occurred during operation. The other two occurred at 1 day and 10 months after stent-graft implantation. Two patients died within 30 days after operation. One died of rupture of the ascending aortic dissection. The other one died of acute renal failure. The 1-month mortality was 3.2%. Four patients underwent a second stent-graft implantation before discharge. One year after stent-graft implantation, complete thrombosis of the false lumen in the thoracic aorta was achieved in 98.4% of patients, and the maximum diameter of the descending aorta decreased 11.2% +/- 7.3%. Three patients died within the follow-up time. Mortality during the follow-up period was 4.8%. One patient died of peptic ulcer hemorrhage. Another one died of rupture of the ascending aortic dissection. The third one died of unknown reasons. The actuarial survival curve by the Kaplan-Meier method showed a 4-year survival rate of 89.4%. CONCLUSIONS: Early and mid-term results showed that endovascular repair was effective in treatment of acute type B aortic dissection. With the enrichment of doctors' experience and refinement of the device, better results are expected in the future.


Assuntos
Aneurisma Aórtico/cirurgia , Dissecção Aórtica/cirurgia , Implante de Prótese Vascular/métodos , Stents , Doença Aguda , Adulto , Idoso , Idoso de 80 Anos ou mais , Dissecção Aórtica/diagnóstico por imagem , Aneurisma Aórtico/diagnóstico por imagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Taxa de Sobrevida , Tomografia Computadorizada por Raios X , Resultado do Tratamento
19.
Zhongguo Zhong Yao Za Zhi ; 31(5): 419-21, 2006 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-16711432

RESUMO

OBJECTIVE: To observe the antiarthritic effects and the possible mechanism of total saponins of Psammruosilene tunicoids (TSPT) against rheumatoid arthritis (RA). METHOD: After establishing AA rat model, the TSPT'S antiarthritic effects and mechanism against RA were studied through observing the changes of ankle swelling, arthritis index and levels of IL-1beta and TNF-alpha after medication. RESULT: TSPT could effectively inhibits articular swelling, decrease arthritis index and regulate down the content of IL-1beta and TNF-alpha in the inflammatory tissue soak of AA rats. CONCLUSION: TSPT has good antiarthritic effects and the possible mechanism may be related to its down-regulation of IL-1beta and TNF-alpha.


Assuntos
Artrite Reumatoide/metabolismo , Caryophyllaceae , Saponinas/farmacologia , Animais , Artrite Reumatoide/patologia , Caryophyllaceae/química , Pé/patologia , Interleucina-1/metabolismo , Masculino , Raízes de Plantas/química , Plantas Medicinais/química , Ratos , Ratos Wistar , Saponinas/isolamento & purificação , Fator de Necrose Tumoral alfa/metabolismo
20.
Ann Thorac Surg ; 80(3): 864-8, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16122444

RESUMO

BACKGROUND: The purpose of this study is to evaluate the feasibility and safety of stent graft in the treatment of aortic dissection and penetrating aortic ulcer. METHODS: From June 2001 to April 2004, 25 patients with aortic dissection and 5 patients with penetrating aortic ulcer received stent-graft implantation. Within this group were 24 male and 6 female patients, with an average age of 52.3 +/- 11.9 years. One patient was diagnosed as type A dissection, 23 patients as chronic type B dissection, 1 patient as acute type B dissection, and the remaining 5 patients with penetrating aortic ulcer. Among the 25 patients with aortic dissection, 3 had contained rupture. All patients were followed up in 1 to 32 months. RESULTS: All patients received stent-graft implantation. There were 5 type I endoleaks. Retrograde ascending aortic dissection occurred during the operation in 1 patient. Two other retrograde ascending aortic dissections occurred in 2 patients 1 day and 7 days, respectively, after stent-graft implantation. Two patients died 1 day and 18 days, respectively, after operation because of rupture of ascending aortic dissection. One-month mortality rate was 6.7%. No death occurred during follow-up time. One patient received a second stent-graft implantation 20 months after the first procedure. CONCLUSIONS: Serious complications may develop after stent-graft implantation, but the early results of treatment of aortic dissection and penetrating aortic ulcer with stent graft were satisfactory. Long-term follow-up was needed.


Assuntos
Aneurisma Aórtico/cirurgia , Dissecção Aórtica/cirurgia , Úlcera/cirurgia , Adulto , Idoso , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Procedimentos Cirúrgicos Cardíacos/métodos , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Recidiva , Stents , Transplantes , Resultado do Tratamento
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