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OBJECTIVE To study the composition of chemical constituents of Sargassum fusiforme and its in vitro anti- neuroinflammatory activity ，and to provide reference for its development and utilization and the study of pharmacodynamic substances. METHODS UHPLC-QTOF-MS/MS analysis method and GC-MS/MS method were used to analyze the chemical constituents of S. fusiforme . The lipopolysaccharide （1 μg/mL）was adopted to establish the inflammatory model of neuromicroglia BV2. Using paroxetine （5 μg/mL）as positive control ，CCK-8 assay was used to detect the effects of the extracts of S. fusiforme （20，40，60，80，100 μg/mL）on the activity and morphology of neuromicroglia BV 2. The effects of the extracts of S. fusiforme （40，60，80 μg/mL）on the contents of tumor necrosis factor α（TNF-α）and interleukin- 6（IL-6）in cell supernatant were detected by ELISA. RESULTS A total of 103 non-volatile constituents were identified by UHPLC-QTOF-MS/MS ，and 60 volatile constituents were obtained by GC-MS/MS. The extracts of S. fusiforme （40，60，80 μ g/mL） could significantly reduce the abnormally increased activation of neuromicroglia BV 2 and the contents of TNF-α and IL-6 due to lipopolysaccharide （P＜0.05 or P＜0.01）. CONCLUSIONS The study establish the full spectrum of chemical constituents of S. fusiforme ，and it is confirmed that fusiforme has certain in vitro anti-neuroinflammatory activity.
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Sargassum fusiforme (S. fusiforme) has been used as an ingredient in Chinese herbal medicine for thousands of years. However, there are a limited number of studies concerning its therapeutic mechanism. High performance gel permeation chromatography (HPGPC) analysis showed that the average molecular weight of the S. fusiforme polysaccharide, SFPS 191212, is 43 kDa. SFPS 191212 is composed of mannose, rhamnose, galactose, xylose, glucose, and fucose (at a molar ratio: 2.1 : 2.9 : 1.8 : 15.5 : 4.6 : 62.5) with α- and β-configurations. The present research evaluated the anti-tumor potential of the S. fusiforme polysaccharide in human erythroleukemia (HEL) cells in vitro. To explore the SFPS 191212's apoptosis mechanism in HEL cells, transcriptome analysis was performed on HEL cells that were incubated with SFPS 191212. The inhibitory effect of SFPS 191212 on HEL cell growth was also analyzed. It was found that SFPS 191212 inhibited HEL cell proliferation, reduced cell viability in a concentration-dependent manner, and induced an insignificant toxic effect on normal human embryonic lung (MRC-5) cells. Compared with the control group, transcriptome analysis identified a total of 598 differentially expressed genes (DEGs), including 243 up-regulated genes and 355 down-regulated genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on all DEGs, and 900 GO terms and 52 pathways were found to be significantly enriched. Finally, 23 DEGs were randomly selected and confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). Moreover, SFPS 191212 down-regulated the PI3K/Akt signal transduction pathway. Our results provide a framework for understanding the effect of SFPS 191212 on cancer cells and can serve as a resource for delineating the anti-tumor mechanisms of S. fusiforme.
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Humans , Leukemia, Erythroblastic, Acute , Phosphatidylinositol 3-Kinases , Polysaccharides/pharmacology , Sargassum , TranscriptomeABSTRACT
This study aimed to investigate the effects of Sargassum fusiforme polysaccharide (SFPS I, II, and III) on the apoptosis and regulation of human erythroleukemia (HEL) cells. The effect of different doses of SFPS on HEL cell growth was detected using the Cell Counting Kit-8 method, and apoptosis was detected by Hoechst staining. Cell cycle distribution and apoptosis were detected using flow cytometry. Expression of the cell cycle gene, p53, antiapoptotic genes, Bcl-xL and Bcl-2, and pro-apoptotic genes, Bax, Bad, and Caspase-3, as well as the expression of the corresponding proteins, were detected using real-time quantitative polymerase chain reaction (qPCR) and Western blot. The results showed that SFPS II and III decreased HEL cell viability and induced HEL cell apoptosis. Different concentrations of SFPS (I, II, and III) were detected that induced much less toxic effect in normal human embryonic lung (MRC-5) cells, and SFPS I increased cell proliferation, indicating its favorable selectivity towards cancer cells. The mechanism by which SFPS induced apoptosis was also found to be related to the induction of cell cycle arrest in the G/G phase and the increased expression of apoptosis-related genes and proteins. We concluded that SFPS induces HEL cell apoptosis, possibly via activation of the Caspase pathway, providing the theoretical basis for the development of SFPS-based anti-tumor drug products.
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Objective: To study the chemical constituents of the whole herbs of Sargassum fusiforme. Methods: The chemical constituents from S. fusiforme were separated and purified by silica gel, MCI, ODS, Sephadex LH-20 gel column chromatographies, and preparative HPLC. Their structures were identified by analysis of NMR, MS, ORD, etc, and comparing physicochemical properties of compounds with references. Results: A total of 19 compounds were obtained from the 50% ethanol-H2O extract of S. fusiforme, and their structures were determined as lupenone (1), bauerenone (2), β-amyrenone (3), α-amyrenone (4), loliolide (5), isololiolide (6), (3S,5R,6S,7E)-5,6-epoxy-3-hydroxy-7-megastigmen-9-one (7), (R)-dehydrovomifoliol (8), 24-ethylcholesta-4,24 (28)-dien-3-one (9), fucosterol (10), 29-hydroperoxystigmasta-5,24 (28)-dien-3β-ol (11), 24-hydroperoxy-24-vinylcholesterol (12), saringosterol (13), (24R)-5,28-stigmastadiene-3β,24-diol-7-one (14), dihydroquercetin (15), aurantiamide acetate (16), dibutyl phthalate (17), glycerol monopalmitate (18), and methyl hexadecanoate (19). Conclusion: The compounds isolated from S. fusiforme were mainly identified as triterpenoids, sesquiterpenoids, highly oxidized sterols, alkaloids and so on. Compounds 1-4, 7-8 and 14-19 are isolated from S. fusiforme for the first time.
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Objective To study the mechanisms of anti-oxidation and anti-aging of Sargassum fusiforme polysaccharides (SFPS). Methods qRT-PCR and Western blotting were utilized to detect the expression of c-Jun-N-terminal kinase 1/2 (JNK1/2) in male mice with different ages and in the old male mice fed with SFPS. The JNK isoforms were specifically distinguished by RT-PCR and the expression levels of each isoform in the liver of mice by ig administration of SFPS or normal saline were measured. Finally, the mechanism of JNK activating Nrf2/ARE signaling pathway was determined by co-immunoprecipitation and ARE-luciferase reporter gene assay. Results The expression levels of JNK1/2 were negatively correlated with aging process. JNK1-β2, JNK2-α1, and JNK2-β2 played important roles in aging process. The mRNA expression level of JNK1-β2, JNK2-α1, and JNK2-β2 were significantly decreased in mice aging process, and the protein expression levels of JNK1-β2 (5.4×104) were significantly upregulated by long-term diet with SFPS. The JNK1-β2 (5.4×104) protein interacted with anti-oxidant factor Nrf2, and significantly activated Nrf2/ARE signaling pathway. Conclusion JNK1-β2, JNK2-α1 and JNK2-β2 had negative correlation with aging process, and the expression level of JNK1-β2 (5.4×104) in aged mice were obviously promoted by long-term diet with SFPS. As JNK1-β2 containing C-terminal tail can interact with Nrf2 and activate the NRF2/ARE signaling pathway, therefore, it was suggested that SFPS can improve the overall anti-oxidant capacity and retard aging process by activating of the JNK1-β2/Nrf2/ARE signaling pathway.
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Objective To indentify monogalactosylmonoacylglycerol (MGMG) and analyze the chemical characteristics of sn-1 and sn-2 type isomers in Marine TCM Sargassum fusiforme. Methods The MGMG compounds were isolated by multiple chromatography methods and their structures were determined by spectroscopic methods such as NMR and HRESI-MS. The interconversion and chromatography-mass spectrometry feature of MGMG positional isomers were analyzed through HPLC and HPLC-ESI-MS/MS. Results Five MGMGs were isolated from Sargassum fusiforme, and the fatty acyl compositions were stearidonoyl, linolenoyl, arachidoyl, linoleyl, and oleyl. The sn-1 and sn-2 type of MGMG were unstable. The interconversion tended to be sn-1 type rapidly, and kept balanced content at 85:15. The relative retention time of sn-2 type was less than sn-1 type on C18 reverse phase chromatography. The abundant fragment ions of MS/MS were significantly differences. The base peak was [M+Na-Gal]+ (100%), and the peak intensity of [M+Na-RCOOH]+ was always more than 50% from sn-2 type while less than 20% from sn-1 type. Conclusion Five MGMGs were isolated from brown algae for the first time. This was the first report about the interconversion and chromatography-mass spectrometry feature of sn-1 and sn-2 type. It can use to determine the fatty acyl attachment in each MGMG.
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Objective:To establish a GC-MS method for determing 19 organochlorine pesticide residues in sargassum fusiforme. Methods:Nineteen organochlorine pesticide residues were simultaneously determined by GC-MS, and the established method was vali-dated. The HP-5 gas chromatography column (30 m × 0. 25 mm,0. 25 μm) was used. The column temperature was programming in-creased with initial temperature of 70℃, maintaining for 1min, then raising to 180℃ with a rate of 15℃·min-1 ,and then raised to 280℃ with a rate of 4℃·min-1 to keep 7min. The inlet temperature was 240℃. The MS detector was used with EI source at 230℃, and analysis mode was multiple reaction monitoring. Results:Sargassum fusiforme was analyzed. The separation degree of the standard and samples met the requirements. The recoveries of the organochlorine pesticides were 72%-127% except for hexachlorobenzene, and the relative standard deviation also met the requirements. The experimental results showed that the 19 organochlorine pesticides were not detected out in sargassum fusiforme. Conclusion: The method provides the technical parameters for the determination of organo-chlorine pesticide residues in sargassum fusiforme.
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AIDS is a global pandemic that has seen the development of novel and effective treatments to improve the quality of life of those infected and reduction in the spread of the disease. While great advancements have been made in HIV/AIDS therapeutics, there is still no cure or viable vaccine in development. The high rate of HIV-1 mutation contributing to virus immune escape, combined with increase in sexual transmission and the significant clinical therapeutics side effects of the currently available treatments highlights the need for novel therapeutics with broad anti-viral activity against both CXCR4 (X4) and CCR5 (R5)-tropic viruses. In our search for novel modalities against HIV infection, we investigated several aqueous extracts from Traditional Chinese Medicine (TCM) collection and identified Sargassum fusiforme (S. fusiforme) as a potent inhibitor of HIV infection. Following the Western approach of drug discovery and development, we isolated several bioactive molecules from S. fusiforme and determined their mechanism of action. TCM and Western approaches to disease treatment and drug development have been shown to be complementary to each other. The former provides a rich medicinal history for natural compounds that can have clinical impact on a variety of illnesses, while the latter enables the application of chemical informatics with rational drug design approach coupled to mechanistic underpinnings of such therapeutics. This multistep paradigm is demonstrated in the example of S. fusiforme as a treatment to prevent HIV infection, as described in this review.
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Sargassum fusiforme has traditionally been widely consumed in Asia as a food, and it has gained much attention due to its high nutritional, pharmaceutical, and industrial value. This study aimed to examine the promotional effects of ethanol extract (ET) and fraction obtained from ethyl acetate (FR) of S. fusiforme on hair growth in C57BL/6 mice and HaCaT cells. Five-week-old mice were used to compare hair regrowth during application of ET and FR for 21 days. Hair regrowth was evaluated by macroscopic observation and verified by hematoxylin-eosin tissue staining. Levels of mRNA expression of factors relevant to the hair growth cycle such as keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), and transforming growth factor-beta1 (TGF-beta1) were examined by quantitative polymerase chain reaction (qPCR). Our results showed that ET and FR successfully promoted hair regrowth in shaved C57BL/6 mice at a dose >20 mg/kg. Moreover, ET and FR were effective in stimulating expression of KGF and VEGF mRNAs in a dose-dependent manner, whereas TGF-beta1 was not activated. These results indicate that ET and FR of S. fusiforme effectively promoted hair growth and gene expression relevant to hair growth cycles in both in vitro and in vivo models.
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Animals , Mice , Alopecia , Asia , Ethanol , Fibroblast Growth Factor 7 , Gene Expression , Hair , Polymerase Chain Reaction , RNA, Messenger , Sargassum , Transforming Growth Factor beta1 , Vascular Endothelial Growth Factor AABSTRACT
Objective To study the anti-tumor effects of SFPS against human colon cancer cells.Methods Inhibition of the cell proliferation was measured by MTT assay.SFPS induced apoptosis of lovo and RKO cells was observed by electron microscopy and flow cytometry.The potential of SFPS in inhibiting lovo and RKO cells viability was assessed by MTT assay.Results SFPS significantly exhibited antiproliferative activity which depended on dosage.Morphological examination showed chromosomal condensation, karyotheca margination,cell shrinkage and the presence of apoptosis bodies.The overall effect of SFPS on the cell cycle distribution was examined by flow cytometry.However,it was also found that SFPS arrested the human colon cancer cell line RKO at G0/G1 phase,and the RKO cells at S phase decreased significantly,while no change in cell cycle distribution from SFPS treated lovo cells was observed.Conclusion SFPS may induce the apoptosis of lovo and RKO cells in vitro through anti-tumor proliferation.
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Objective To investigate the effects of polysaccharide from Sargassum fusiforme on blood-lipid and antioxidation ability in hyperlipemia rats.Methods The hyperlipemia rats as the experimental animal were established by feeding with fat-rich forage,meanwhile the rats were ig SFPS at doses of 0.4、0.8、1.6g?kg-1for 4wk.The levels of blood-lipid and the indices of antioxidation ability which include the total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),lipid peroxidation(LPO),malonaldehyde(MDA),superoxide dismutase(SOD) and glutathion peroxidase(GSH-Px) were determined.Results and Conclusion SFPS could significantly reduce the contents of TC,TG,LDL-C,LPO and MDA.In the mean time,SFPS increased the content of HDL-C and enhanced the activities of SOD and GSH-Px.
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Object To study the inhibitory effect of Sargassum fusiforme (Harv.) Setch. polysaccharide (SFPS) on six different kinds of tissue tumor by antitumor experiments in vitro, and to observe tissue speciality of tumor treatment and reveal antitumor effect mechanism. Methods MTT and colony forming methods were adopted to study the antitumor activity of SFPS in vitro. Influences of SFPS on cell cycle and apoptosis were observed. LCM was adopted to measure [Ca 2+ ]i of the cells marked by Fluo 3/AM probe. Results SFPS had content antitumor activities to SGC 7901 and COLO 205. It prevented the transforming of SCG 7901 from G 0/G 1 period to S period. And it increased the apoptosis index (APO%). Applying SFPS, [Ca 2+ ]i increased first and then decreased. Further, after giving CaCl 2, [Ca 2+ ]i increased again. Conclusion By inducing the apoptosis of tumor cells, SFPS shows content antitumor activities to SGC 7901 and COLO 205. SFPS could increase the [Ca 2+ ]i in SGC 7901. And the Ca 2+ are from cellular storage.
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Objective Bioactive components were extracted from Sargassum fusiforme by using different solvents and their activities were detected in vitro in order to screen potential free radical scavenger.Methods Using the assay system of DPPH,superoxide radical,and in comparison with vitamin C,tea polyphenols and a series of extracts of sea tangle,laver.Results Different solvents had different extraction rates,and extracts by different solvents had different free radical scavenging activities.Conclusion 0.5%Na2CO3 solution had the highest extraction rate and extracts by this solution had the best scavenging ability as well.At the same time,the results obtained by DPPH detection method were more stable than those by the others.
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ObjectiveTo investigate the inhibitory effect of polysaccharides from Sargassum fusiforme on the HepG 2 cells. Methods Six kinds of polysaccharides were prepared by hot water extracting, ethanol and CaCl 2 precipitating and acid hydrolyzing. Inhibition rate of these polysaccharides on the HepG 2 cells were determined by MTT bioassay. Results Inhibition rate of two kinds of polysaccharides increased at lower concentration. Inhibition ratie of other polysaccharides increased with the increment of concentration. At the dose less than 2000mg?L -1,inhibition rate was not more than 40%. Conclusion All polysaccharide specimens obtained from Sargassum fusiforme have some inhibitory effects on human liver cancer cells HepG 2.
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This paper deals with the study on the sexual and vegetative reproduction of Sargassum fusiforme. The authors also propose a inference that Sargassum fusiforme relies mainly on the vegetative reproduction of rhizoid while making sexual reproduction subsidiary to maintain the population propagation.
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Objective To study the chemical constituents of brown alga Sargassum fusiforme.Methods The compounds were isolated by silica gel chromatography,ODS chromatography,and preparative HPLC,and their structures were elucidated on the basis of chemical and spectroscopic methods,including HR EI-MS,1D and 2D NMR spectral techniques.Results Six sterols and two glycolipids were isolated from the n-hexane soluble part of ethanol extract of S.fusiforme and their structures were identified as fucosterol(Ⅰ),24R,28R-and 24S,28S-epoxy-24-ethylcholesterol(Ⅱ),24-hydroperoxy-24-vinylcholesterol(Ⅲ),29-hydroperoxystigmasta-5,24(28)-dien-3?-ol(Ⅳ),(24S)-5,28-stigmastadien-3?,24-diol(Ⅴ),(24R)-5,28-stigmastadien-3?,24-diol(Ⅵ),1-O-myristoyl-3-O-(6′-sulfo-?D-quinovopyranosyl) glycerol(Ⅶ),and 1-O-palmitoyl3-O-(6′-sulfo-?-D-quinovopyranosyl) glycerol(Ⅷ).Conclusion This is the first report for the isolation of Ⅳ,Ⅶ,and Ⅷ from alga of Sargassum(Turn.) Ag.,and compounds Ⅱ and Ⅲ from this alga.The isomers of saringosterols 24S(Ⅴ) and 24R(Ⅵ) from this alga are obtained by preparative normal-phase HPLC for the first time.
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5 000 mg/L). ② All extracts except algin (ALG) had an obviously HSV-1 inhibitory effect. Antiviral activities of them were increasingly strengthened with their purities. Antiviral activities of extracts from flucoidan 1, F 1, F 2 and F 4 were better than acyclovir (ACV). ③ Antiviral activities of all extracts on CVB 3 were better than ribavirin injection. F 1, F 2 and F 4 had remarkable anti-CVB 3 effects. ④ The experiment showed that Sargassum fusiforme polysaccharides not only kill the above viruses directly but also restrain them via getting into cells or absorbing on cells.