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Based on the characteristics and ISSR molecular marker technology, the study is aimed to compare and perform genetic diversity analysis on Sparganium stoloniferum from 7 regions. Molecular identification method was established for S. stoloniferum from Hunan province. Differences among Sparganii Rhizoma samples from seven habitats were analyzed via measuring weight, length, width and thickness of them. Genetic diversity of S. stoloniferum from 7 regions was analyzed by screening out primers amplifying clear band and showing rich polymorphism, then a cultivars dendrogram was built. The target primer was screened out, and the specific band was sequenced. Nine ISSR primers were selected to amplified clear band, rich polymorphism. A total of 73 bands were amplified by nine ISSR primers selected from 27 ISSR primers. On average, each primer produced 8.0 bands. A total of 38 bands were polymorphic, which occupied 52.8% of all bands. The cultivars dendrogram showed the genetic similarity was 0.54-0.94. Genetic similarity coefficient of S. stoloniferum from Jiangsu province, Anhui province and Jiangxi province was big, indicating the differences among them were slight on genetic level. S. stoloniferum from Hunan province is quite different from samples from the other six habitats on appea-rance and genetic level. A specific band(327 bp) in S. stoloniferum from Hunan province was obtained via ISSR-857 primer, and was sequenced. According BLASTn database, there were few sequences similar to the gene fragment and had little correlation with the growth process of plant. ISSR molecular marker technology provides a new idea for the identification of S. stoloniferum. This result confirmed the particularity of S. stoloniferum from ancient Jingzhou.
Subject(s)
China , Drugs, Chinese Herbal , Genetic Markers/genetics , Genetic Variation , Microsatellite Repeats , Phylogeny , Polymorphism, GeneticABSTRACT
OBJECTIVE： To optimize the purification technology of total flavonoids from Sparganium stoloniferum. METHODS： Separation and purification by macroporus adsorption resin， using sample solution pH， flow rate and concentration of eluent， the purification rate of total flavonoids as evalution indexes， the purification technology of total flavonoids from S. stoloniferum were optimized by Box-Behnken design-response surface methodology based on single factor test. Validation test was conducted. RESULTS： The optimal purification technology was sample solution pH 4.8， flow rate of eluent 2.0 BV/h， concentration of eluent 72％. The purification rate of total flavonoids in 3 batches of samples was 72.34％ （RSD＝1.77％， n＝3） in validation test， relative errors of which to predicted value （73.99％） was 2.13％. CONCLUSIONS： The optimal purification technology is stable and feasible， and can be used for the purification of total flavonoids from S. stoloniferum.
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OBJECTIVE:To study the decoction rate changes of 4 active ingredients in the volatile oil of Curcuma phaeocaulis before and after combined with Sparganium stoloniferum,and provide reference for showing the compatibility mechanism of C. pha-eocaulis and S. stoloniferum. METHODS:HPLC was used to simultaneously determine the contents of 4 active ingredients(curdi-one,curcumol,germacrone and β-elemene) in the volatile oil of C. phaeocaulis after C. phaeocaulis single decoction and com-bined decoction with S. stoloniferum. And the fried rates of each ingredient were compared. RESULTS:Compared with C. phaeo-caulis single decoction,there was significant difference in the fried amounts of curdione,curcumol,germacrone in the volatile oil after combined decoction(P0.05). CONCLUSIONS:The com-bined decoction of C. phaeocaulis and S. stoloniferum can effectively improve the dissolution of curdione,curcumol,germacrone in the volatile oil. It may be the reason for the enhancement of efficacy after compatibility.
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Sparganium stoloniferum is widely used in traditional Chinese medicines. It has been reported to exhibit therapeutic effects on thrombus, solid tumor, endometriosis and so on. It is usually used to treat endometriosis and chronic atrophic gastritis, while its underlying mechanisms are poorly delineated. According to vast information from literatures in the last decade, the research on origin of medica, chemical compositions and pharmacological actions were summarized, in hopes of offering more clues for further research as well as clinical application of S. stoloniferum.
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OBJECTIVE:To study the preventive and therapeutic effect and its mechanism of medicine pair decoction liquid of Curcuma phaeocaulis-Sparganium stoloniferum on rats with uterine myoma. METHODS:Rats were randomly divided into normal group,model group,positive group(mifepristone,2.25 mg/kg)and medicine pair decoction liquid group(C. phaeocaulis-S. stolon-iferum decoction liquid,6.0 g/kg),10 in each group. Except for the normal group,rats in other groups were injected Estradiol ben-zoate injection (0.5 mg/kg) intramuscularly every Monday,Wednesday and Friday,for 12 weeks. From the 13th week,rats in modeling group were added Progesterone injection(5 mg/kg)intramuscularly as well as relevant medicines intragastrically,once a day,until the 16th week. After administration,uterine coefficient of rats was detected. HE staining was used to observe the patho-logical changes of uterus and determine the thickness of smooth muscle;immunohistochemical staining was adopted to detect the transforming growth factor β3 (TGF-β3),matrix metalloproteinase 11 (MMP-11) protein expressions in uterus tissue of rats. RE-SULTS:Compared with normal group,uterine coefficient was increased in model group,pathological changes were obvious in uterus,thickness of smooth muscle was increased,TGF-β3 and MMP-11 protein expressions were enhanced(P<0.05 or P<0.01). Compared with model group,above-mentioned changes were improved significantly in positive group and medicine pair decoction liquid groups(P<0.05 or P<0.01). CONCLUSIONS:Medicine pair decoction liquid of C. phaeocaulis-S. stoloniferum shows cer-tain preventive and therapeutic effect on rats with uterine myoma. The mechanism may be associated with downregulating the TGF-β3,MMP-11 protein expressions in uterus tissue.
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ABSTRACT Context The rizoma of Sparganium stoloniferum has been used as a traditional Chinese medicinal herb for thousands of years. Sparganium stoloniferum is a stasis-breaking drug to treat a wide range of diseases including cancer,however, its activity of extract on cervical cancer HeLa cells and the mechanisms remains unknown. Objective This study aimed to screen Sparganium stoloniferum extract for its inhibitory effects on cervical cancer HeLa cells. Materials and methods Sparganium stoloniferum was extracted with 95% ethanol under reflux, and the extracts were preliminary separated by silica gel column chromatography. MTT assay and flow cytometry were used to determine the inhibitory effects of three fractions on HeLa cells. In addition, GC-MS was performed to analyze the chemical composition of the active fraction. Results Sample II showed a dose-dependent inhibition of HeLa cell growth, with an inhibition rate of more than 30%, whereas the inhibition rates of Samples I and III were less than 30%. Interestingly, Samples I and III had no effect on apoptosis, in contrast to Sample II, which significantly promoted HeLa cell apoptosis, in a dose-dependent manner. GC-MS was performed to analyze the chemical components of Sample II: steroids were found to be the major components with a relative content of 73.905%, while six known compounds were obtained for the first time. Discussion and conclusion This study provided a novel insight for further research of active fractions of Sparganium stoloniferum, in accordance with the basic principle and theory of traditional Chinese medicine (TCM), and will promote the shift of effective substance study, from monomeric chemical compounds to active fractions.
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Objective:To explore a new method to identify Sparganium stoloniferum and its adulterants by ITS2 regions. Methods:Eight samples of Sparganium stoloniferum and its adulterants were collected with five species, and 6 species with 23 ITS2 sequence of Sparganium stoloniferum and its adulterants were downloaded from Genbank. The intraspecific and interspecific K2P distances of Spar-ganium stoloniferum and its adulterants were calculated by MEGA5. 0, and the phylogenetic tree was constructed by MEGA 5. 0. Re-sults:The maximum intraspecific K2P distance of Sparganium stoloniferum was 0. 038,while the minimum interspecific K2P distance was 0. 697. The phylogenetic tree showed that Plantago asiatica was different obviously from its adulterants. The different samples of Sparganium stoloniferum were gathered together and could be distinguished from its adulterants by the NJ tree. Conclusion: ITS2 se-quence is able to identify Sparganium stoloniferum and its adulterants correctly, which provides a new method for the identification of Sparganium stoloniferum.
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The optimum conditions for the processing of Sparganium stoloniferum Buch-Ham. withvinegar were studied by L9(3)4 orthogonal experimental design, as guided by decrease of writhing rate inanalgesic test and anticoagulant activity. The results showed that the best processing conditions were stirfrying of the raw S. stoloniferum with the addition of 25 kg of vinegar per 100 kg of crude drug at 130℃for 10 min.
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Objective To study the chemical constituents in Sparganium stoloniferum. Methods The compounds were isolated by repeated silica gel and Sephadex LH-20 chromatographies. The structures of isolated compounds were identified by analysis of their spectral data and chemical reactions. Results Nine compounds were isolated and their structures were identified as daucosterol palmitate (Ⅰ), ?-sitosterol palmitate (Ⅱ), 24-methylenecycloartanol (Ⅲ), 6, 7, 10-trihydroxy-8-octadecenoic acid (Ⅳ), vanillic acid (Ⅴ), p-hydroxybenzaldehyde (Ⅵ), 2,3-dihydroxypropyl palmitate (Ⅶ), 5-hydroxymethyl-2-furaldehyde (Ⅷ), ?-sitosterol (Ⅸ). Conclusion Compounds Ⅰ-Ⅲ and Ⅴ-Ⅷ are obtained from the plants of Sparganium L. for the first time.