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Aim To investigate the effects of Cichorium glandulosum N-butanol extraction site (C G E) on hepatic fibrosis (H F) in SD rats and to determine the content of the main effective component matricin. Methods HPLC method was used to determine the content of matricin in CGE. The SD rats were randomly divided into control group, model group, CGE low-dose groups, medium-dose and high-dose, and curcumin group. In addition to control group rats' back subcutaneous injection (s c) normal saline, rats in the other groups were treated with body weight sc 40 % CC1
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Objective To establish molecular identification method of Cichorium glandulosum and its adulterants Cichorium intybus by Allele-Specific PCR. Methods The samples of C. glandulosum and C. intybus were collected in different geographical areas. The DNA was extracted, and rbcL gene segments were amplified and sequenced directionally. The multiple sequences were aligned by using Clustal W. Specific primers were designed and amplified according to its variable sites, and PCR reaction system was optimized to determine detection limits and establish Allele-Specific PCR identification method. Results According to Allele-Specific PCR system established in this study for C. glandulosum, the optimization results was a total of 30 μL reaction system containing TaqDNA polymerase 0.25 μL, 10 × buffer 2.5 μL, dNTP 2.0 μL, primer 0.5 μL, template DNA 2 μL, and ddH2O 22.25 μL. The most suitable PCR amplification procedure is one cycle of predegeneration at 94 ℃ for 3 min; 32 cycles of denaturing at 94 ℃ for 30 s, annealing at the primer temperature 55 ℃ for 30 s and extending at 72 ℃ for 1 min, and extending at 72 ℃ for 7min. Through the detection of 20 medicinal materials of C. glandulosum and C. intybus, the result showed that 230 bp amplified band of target fragment was identified for C. glandulosum but no amplified band was observed for its adulterants. Conclusion In this study, we established and optimized the Allele-Specific PCR identification technology of C. glandulosum and its adulterants C. intybus, which can accurately, reliably, and effectively identify these two medicinal materials.
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Objective: To analyze the ribosomal DNA internal transcribed spacer (rDNA-ITS) sequence of Cichorium intybus and C. glandulosum from different habitats, and to provide DNA molecular marker for the identification of chicory. Methods: The total DNA was extracted from the samples of C. intybus and C. glandulosum from 14 habitats by rapid broad spectrum of plant genomic DNA extraction kit. The ITS sequence was amplified by PCR with universal primer of ITS and then sequenced. The two kinds of ITS sequences were compared by DNAMAN V6 software. The cluster analysis was adopted by SPSS 17.0 after the different ITS bases from all the samples were mathematically treated. Results: The intraspecies identity of ITS sequence was above 99.2% in C. intybus, and that in C. glandulosum was above 99.8%, while the interspecies identity of ITS sequence was below 99.2%. There were various specific information sites in the ITS sequences of the two kinds of chicory. Conclusion: The ITS sequence is an available molecular marker for the identification of C. glandulosum and C. intybus.