Heat Stress and Microbial Stress Induced Defensive Phenol Accumulation in Medicinal Plant Sparganium stoloniferum.

An approach based on the heat stress and microbial stress model of the medicinal plant Sparganium stoloniferum was proposed to elucidate the regulation and mechanism of bioactive phenol accumulation. This method integrates LC-MS/MS analysis, 16S rRNA sequencing, RT-qPCR, and molecular assays to investigate the regulation of phenolic metabolite biosynthesis in S. stoloniferum rhizome (SL) under stress. Previous research has shown that the metabolites and genes involved in phenol biosynthesis correlate to the upregulation of genes involved in plant-pathogen interactions. High-temperature and the presence of Pseudomonas bacteria were observed alongside SL growth. Under conditions of heat stress or Pseudomonas bacteria stress, both the metabolites and genes involved in phenol biosynthesis were upregulated. The regulation of phenol content and phenol biosynthesis gene expression suggests that phenol-based chemical defense of SL is stimulated under stress. Furthermore, the rapid accumulation of phenolic substances relied on the consumption of amino acids. Three defensive proteins, namely Ss4CL, SsC4H, and SsF3'5'H, were identified and verified to elucidate phenol biosynthesis in SL. Overall, this study enhances our understanding of the phenol-based chemical defense of SL, indicating that bioactive phenol substances result from SL's responses to the environment and providing new insights for growing the high-phenol-content medicinal herb SL.

Integration of 16 S rRNA gene sequencing, metabonomics and metagenome analysis to investigate the mechanism of Sparganium stoloniferum-Curcuma phaeocaulis in treating of endometriosis in rats.

BACKGROUND: Endometriosis is a gynecological disease that causes severe chronic pelvic pain and infertility in women. The therapeutic efficacy of the traditional herbal combination of Sparganium stoloniferum-Curcuma phaeocaulis (Sangleng-Ezhu, SL-EZ) in the treatment of endometriosis has been established. However, the precise mechanism by which this treatment exerts its effects remains elusive. METHODS: To gain further insights, UPLC-MS/MS was employed to identify the primary chemical constituents of SL-EZ in serum. Additionally, network pharmacology was utilized to analyze the active ingredients and their corresponding targets. Furthermore, the impact of SL-EZ on ectopic endometrial growth in endometrial implants was assessed using a rat model. The therapeutic mechanism of SL-EZ in rats with endometriosis was further investigated through the application of 16 S rRNA gene sequencing, metagenomic sequencing, and metabolomics. RESULTS: The primary compounds in serum were zederone, p-coumaric acid, dehydrocostus lactone, curdione, curcumol. The growth of ectopic lesions in a rat model was effectively inhibited by SL-EZ. In comparison to the control group, the endometriotic rats exhibited a decrease in α-diversity of the gut microbiota, an increase in the Firmicutes/Bacteroidetes ratio, and a reduction in the abundance of Ruminococcaceae. Following SL-EZ intervention, the potential probiotic strains Lactobacillus gasseri and Lactobacillus johnsonii were able to restore the intestinal microenvironment at the species level. The altered metabolites were significantly correlated with Verrucomicrobia, Proteobacteria, and Bacteroidetes. The metabolomic analysis demonstrated significant alterations in intestinal metabolites. And SL-EZ intervention also exerted regulatory effects on various metabolic pathways in gut microbiota, including aminoacyl-tRNA biosynthesis, monobactam biosynthesis, cyanoamino acid metabolism, glycine, serine and threonine metabolism, plant secondary metabolite biosynthesis, and amino acid biosynthesis. CONCLUSION: The identification of novel treatment formulations for endometriosis was achieved through the utilization of network pharmacology and gut microbiota analyses. Our findings revealed simultaneous alterations in the microbiota within the rat model of endometriosis. The therapeutic efficacy of SL-EZ in treating endometriosis is attributed to its ability to restore the gut microbiota and regulate metabolism. This investigation offers valuable insights into the therapeutic mechanisms of traditional Chinese medicine (TCM) for endometriosis.

Phenylpropanoids from Sparganium stoloniferum and their antiplatelet aggregation activities.

Four new ferulic acid sucrose esters, ß-D-(1-O-acetyl-6-O-trans-feruloyl)fructofuranosyl-a-D-2',6'-O-diacetylglucopyranoside (1), ß-D-(1-O-acetyl-6-O-trans-feruloyl)fructofuranosyl-a-D-2',4'-O-diacetylglucopyranoside (2), ß-D-(6-O-trans-feruloyl)fructofuranosyl-a-D-2',4',6'-O-triacetylglucopyranoside (3), ß-D-(1-O-acetyl-6-O-trans-feruloyl)fructofuranosyl-a-D-4',6'-O-diacetylglucopyranoside (4), together with four known phenylpropanoids (5-8) were isolated from the rhizome of Sparganium stoloniferum (Graebn.) Buch.-Ham. ex Juz. Their structures were elucidated by chemical evidence and spectral analysis. Compounds 1-8 exhibited obvious inhibitory effects on ADP-induced platelet aggregation.

Characterization of the complete chloroplast genome of an aquatic plant, Sparganium stoloniferum subsp. choui (Typhaceae).

Sparganium stoloniferum subsp. choui is an aquatic perennial herb distributed in Northeast China. It was published as a new species in 1992 and recognized as a subspecies of S. stoloniferum in Flora of China in 2010. The complete chloroplast genome of S. stoloniferum subsp. choui was sequenced and assembled. The genome size was 161,865 bp in length with 36.8% GC content. Its quadripartite structure consisted of the large single copy (LSC, 88,953 bp) and small single copy (SSC, 19,098 bp) regions, separated by a pair of inverted repeats (IRs) of 26,907 bp. The genome contained 114 distinct genes, including 80 protein-coding genes, 30 tRNA genes, and four rRNA genes. The phylogenetic analysis including four Sparganium species showed that two subspecies of S. stoloniferum were not monophyletic, supporting the resurrection of S. choui as a species.

[Study on Sparganium stoloniferum from different regions in China based on appearance characters and ISSR markers].

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.

Sparganii Rhizoma: A review of traditional clinical application, processing, phytochemistry, pharmacology, and toxicity.

ETHNOPHARMACOLOGICAL RELEVANCE: Sparganii Rhizoma (SR), a traditional Chinese medicine (TCM), is the rhizome of Sparganium stoloniferum Buch.-Ham. mainly distributed in East Asia. It has been used for eliminating blood stasis, promoting the flow of Qi, removing the retention of undigested food and relieving pain in China for hundreds of years. AIM OF THE REVIEW: This review summarizes comprehensive information in traditional clinical application, processing, phytochemistry, pharmacology, quality control and toxicity of SR, in exploring future scientific and therapeutic potentials. MATERIALS AND METHODS: Pertinent information was systematically collected from several electronic scientific databases (e.g., Web of Science, PubMed, China Knowledge Resource Integrated, Springer, Elsevier, ScienceDirect, and Google Scholar), PhD and MS dissertations, and classic Chinese medical books. RESULTS: SR is a gynecological drug which is often used to treat dysmenorrhea, mass in the abdomen, amenorrhea due to blood stasis, and abdominal distension in TCM. Two kinds of processed products of SR are included in Chinese Pharmacopoeia, which have better pharmacological effects than the crude herb. Approximately 180 compounds have been identified from SR, including phenylpropanoids, flavonoids, anthraquinones, organic acids, alkaloids, steroids, volatile oils, diarylheptanes, etc. The crude extracts and isolated components of SR have been reported to have anti-tumor, antithrombotic, estrogen antagonistic , anti-inflammatory, analgesic, antioxidant, anti organ fibrosis and other pharmacological activities. SR also has reproductive toxicity. CONCLUSIONS: As an important TCM, SR has been demonstrated by modern pharmacological researches to have significant bioactivities, especially on anti-tumor, antithrombotic, and estrogen antagonistic activities. These activities provide prospects for the development of new drugs and therapeutics for future applications. Nevertheless, quality control and evaluation, in-depth pharmacological mechanism, and toxicological effect of SR require further detailed research.

Study on Sparganium stoloniferum from different regions in China based on appearance characters and ISSR markers / 中国中药杂志

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.

Study on the Separation and Purification Technology of Total Flavonoids from Sparganium stoloni by Box-Behnken Design- response Surface Methodology / 中国药房

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.

Two new sucrose esters from the rhizome of Sparganium stoloniferum Buch.-Ham.

Two new sucrose esters, ß-D-(1-O-acetyl-3,6-O-trans-diferuloyl)fructofuranosyl-α-D-2'-O-acetylglucopyranoside (1) and ß-D-(1-O-acetyl-3-O-cis-feruloyl-6-O-trans-feruloyl)fructofuranosyl-α-D-2',4',6'-O-triacetylglucopyranoside (2), together with four known sucrose esters (3-6) have been isolated from the rhizome of Sparganium stoloniferum Buch.-Ham. Their structures were elucidated by physical and chemical evidence and spectral analysis.

New isocoumarin and stilbenoid derivatives from the tubers of Sparganium stoloniferum (Buch.-Ham.).

A new isocoumarin derivative 8,5'-dihydroxy-6'-methoxy-4-phenyl-5,2'-oxidoisocoumarin (1) and a new stilbenoid derivative methyl 5-hydroxy-2-(2-hydroxyphenyl)benzofuran-4-carboxylate (2) together with nine known compounds (3-11) were isolated from the tubers of Sparganium stoloniferum Buch.-Ham.. Another stilbenoid derivative (3) and a xanthone (4) were identified as new natural products and compounds 5-10 were obtained for the first time from the genus Sparganium. All their structures were elucidated by comprehensive spectroscopic analysis and comparison with available literature information.

Research progress on chemical constituents and pharmacological activities of Sparganium stoloniferum / 中草药

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.

Study on the Preventive and Therapeutic Effect and Its Mechanism of Medicine Pair Decoction Liquid of Curcuma phaeocaulis-Sparganium stoloniferum on Uterine Myoma in Rats / 中国药房

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.

Comparative Study on Decoction Rates of Active Ingredients in Volatile Oil after Curcuma phaeocaulis Sin-gle Decoction and Combined Decoction with Sparganium stoloniferum / 中国药房

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.

Two new phenolic compounds from the tuber of Sparganium stoloniferum.

Two new phenolic compounds, 2-(2-hydroxyphenyl)-4-methoxycarbonyl-5-hydroxybenzofuran (1) and 1-methoxycarbonyl-2, 3-dihydroxydibenzo[b, f]oxepine (2), were isolated from the tuber of Sparganium stoloniferum. The structures of both new compounds were determined on basis of spectroscopic means including HR-ESI-MS, 1D and 2D NMR experiments.

Molecular Identification of Sparganium Stoloniferum and its Adulterants Based on ITS2 Sequence / 中国药师

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.

Identification of a fat-soluble fraction of Sparganium stoloniferum that inhibits cervical cancer HeLa cells

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.

Optimization of Processing Technique of Sparganium stoloniferum by Orthogonal Experimental Design / 中草药

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.

Chemical constituents of Sparganium stoloniferum (Ⅰ) / 中草药

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.