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Objective: To investigate the osteoblastogenic activity of the ethyl acetate (EtOAc) extract of Smilax glabra Roxb roots and its major active compound astilbin. Methods: Astilbin was isolated from EtOAc extract using silica gel chromatography combined with fraction crystallization. Chemical structure of astilbin was determined by analysis of the spectroscopic data in comparison with the literature. MTT method was used to detect the toxicity. Alkaline phosphatase (ALP) activity was determined by the spectrophotometric method at 405 nm using p-nitrophenyl phosphate as a substrate. Calcium deposition was stained with alizarin red-S, distained with cetylpyridium chloride, and quantified at 562 nm. In silico model for astilbin-ALP interaction was analyzed using AutoDock 4.2.6. The changes in expression of osteoblast differentiation related genes were determined using quantitative real-time PCR. Results: Both the EtOAc extract and astilbin had no toxicity toward osteoblast MC3T3-E1 cells at 5.0, 10, 25, and 50 μg/mL. At 25 μg/ mL, they enhanced ALP activity and mineralization of osteoblasts up to 30% and 55% for the EtOAc extract and 22% and 41% for astilbin, respectively. Molecular docking analysis of astilbin-ALP interaction revealed Arg167, Asp320, His324, and His437 were key residues participating in hydrophobic interaction; meanwhile, His434 and Thr436 residues were involved in hydrogen bond formation in the active site of human tissue-nonspecific ALP. Moreover, the expression level of genes opn, col1, osx, and runx2 were up-regulated in astilbin treated samples with the fold changes as 2.2; 3.7; 4.1; 2.3, respectively at 10 μg/mL (P<0.05). Conclusions: The EtOAc extract and its major compound astilbin exhibit osteoblastogenic activity by up-regulating important markers for bone cell differentiation. It could be a new and promising osteogenic agent with dual actions for therapeutic applications.
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OBJECTIVE@#The present work tested organic solvents to prepare an extract with anticancer properties from a polyherbal mixture containing Nigella sativa (seeds), Hemidesmus indicus (roots) and Smilax glabra (rhizomes). We evaluate anticancer effects in non-small-cell lung cancer cells (NCI-H292), and discuss optimization for pharmaceutical use in the context of efficacy, yield and toxicity.@*METHODS@#Using different organic solvents, six extracts were prepared from the polyherbal mixture. Based on the cytotoxic effects of these extracts on NCI-H292 cells and normal lung cells (MRC-5), as evaluated by the sulphorhodamine B assay, the total ethyl acetate (T-EA) extract was selected for further analysis. The possible anticancer mechanisms were assessed by evaluating the extract's effects on apoptosis (through fluorescent microscopic analysis, DNA fragmentation analysis, caspase 3/7 assay and analysis of expression levels of apoptosis-related genes p53, Bax, survivin, Hsp70 and Hsp90), colony formation and antioxidant activity.@*RESULTS@#The extract had cytotoxic effects against NCI-H292 cells in a time- and dose-dependent manner. Significant antioxidant activity and inhibition of colony formation were also observed. The expression level of caspase 3/7 significantly (P < 0.001) increased in NCI-H292 cells treated with 50 μg/mL of the extract. The same dosage led to a significant increase in expression levels of Bax and p53 (P < 0.05 and P < 0.01 respectively), accompanied by a significant decrease (P < 0.0001) in survivin, Hsp70 and Hsp90.@*CONCLUSION@#T-EA extract of the above polyherbal mixture has cytotoxicity against NCI-H292 cells via induction of apoptosis, antioxidant effects and inhibition of colony formation.
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Objective: To predict the targets and pathways for the main active components of Smilax glabra in the treatment of gout based on network pharmacology. Methods The active components and targets of the S. glabra were obtained by TCMSP database and Drugbank database. Furthemore, the interaction network among the targets was established by Cytoscape software. Meanwhile, crosslink analysis was performed to screen out the active components and potential targets. Finally, the information was obtained from the MAS 3.0 biomolecular function system, and then the target pathway network model was established. Results In this study, a total of 11 effective components and 39 effective targets were predicted, which related to adipocytokine signaling pathway, ERbB signaling pathway, and Toll like receptor signaling pathway. Among these pathways, MAPK1, RELA, PTGS2 genes may play a crucial role. Conclusion This study investigated the characteristics on multi-component, multi-target and multi-pathway of S. glabra, which provided a new idea and method for further study on anti-gout mechanism of S. glabra.
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Objective: To establish a method of quantitative analysis of multi-components by single-marker (QAMS) for simultaneously determination of neoastilbin, astilbin, neoisoastilbin, isoastilbin, and engeletin in Smilax glabra, and research the tendency of content changes in different growth years by QAMS. Methods: An HPLC method was established to determine the relative correction factors of four other flavonoids by using astilbin as the internal reference standard. Then the method was used to determine the various content of five flavonoids in different growth years and validate the feasibility and accuracy by comparing the content results determined by the external standard method with QAMS. The dynamic change regularity of flavonoids in S. glabra was investigated. Results: A total of 23 samples from five batches in different growth years were simultaneously determined by external standard method and QAMS, the results deviation were all less than 1.0%. The content of five flavonoids in different growth years was different, astilbin decreased with the increase of growth years, neoastilbin, neoisoastilbin and isoastilbin had the reverse trend. Conclusion: The established QAMS method is feasible and accurate for the simultaneous determination of five flavonoids in S. glabra. Growth years have a certain impact on the content of each component.
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Objective: To analyze and compare the volatile components in Smilax glabra Robx. and its adulterant Smilax china L. . Methods: The volatile components in Smilax glabra Robx. and Smilax china L. were analyzed by head space solid phase micro-extraction (HS-SPME) combined with gas chromatography-mass spectrometry ( GC-MS). The relative percentage of each component was calculated by area normalization method. Results: Totally 24 components were detected out from Smilax glabra Robx. , and among them, 20 components were identified, which accounted for 97. 04% of the volatile components. Totally 21 components were detected out from Smilax china L. , and among them, 15 components were identified, which accounted for 77. 76% of the volatile components. Conclusion: The composition and content of volatile components in Smilax glabra Robx. and its adulterant Smilax china L. are differ-ent. The method can provide scientific basis for the identification of Smilax glabra Robx. and its adulterants.
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OBJECTIVE: To investigate the chemical differences between Smilacis Glabrae Rhizoma and its adulterants, and provide a basis for the identification and quality evaluation of Tufuling samples purchased from the markets. METHODS: The method of HPLC fingerprinting was used to analyze Smilacis Glabrae Rhizoma, Heterosmilacis Chinensis Rhizoma, Heterosmilacis Yunnanensis Rhizoma, and 102 Tufuling samples which were collected around China. The fingerprints were analyzed by the methods of similarity and principal component analysis (PCA). RESULTS: The analytical method of HPLC fingerprinting was established. Eleven, fifteen and eight common peaks were selected in the fingerprints of Smilacis Glabrae Rhizoma, Heterosmilacis Chinensis Rhizoma, and Heterosmilacis Yunnanensis Rhizoma, respectively. Only five common peaks were found in the fingerprints of the three species, which were No. 1, 2, 3, 13, and 14 peaks. A total of twelve peaks were characterized in the three fingerprints. Nine peaks were characterized in the fingerprint of Smilacis Glabrae Rhizoma, among which, four constituents were characterized for the first time. Six and two constituents were for the first time characterized in the fingerprints of Heterosmilacis Chinensis Rhizoma and Heterosmilacis Yunnanensis Rhizoma, respectively. The result of PCA analysis showed that the chemical differences between the three species were quite obvious and they could be distinguished from each other. The established method was used for the analysis of Tufuling samples purchased from the markets. Sixty-five samples were identified as Smilacis Glabrae Rhizoma, seventeen samples were identified as Heterosmilacis Yunnanensis Rhizoma, and twenty samples were identified as Heterosmilacis Chinensis Rhizoma. CONCLUSION: The established method is simple and reliable, and the development of fingerprint and its chemical pattern recognition provide the way and basis for identification of Smilacis Glabrae Rhizoma and its adulterants.
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Objective To explore the effects of different extraction methods on yield of Smilax glabra polysaccharides and antioxidant activities. Methods Crude polysaccharides were extracted by using cold water, hot water, NaOH aqueous solution, and EDTA?2Na aqueous solution . Antioxidant activities of Smilax glabra polysaccharides were compared by the methods of scavenging superoxide anion radical , scavenging hydroxyl radical or reducing power. Results Yield of crude cold water-soluble polysaccharides, crude boiling water-soluble polysaccharides, crude alkali-soluble polysaccharides and crude acid-soluble polysaccharides were 0.31%, 1.1%, 10.8 %, 2.0%, respectively. Polysaccharides by using four kinds of different extraction methods had strong scavenging superoxide anion radical power; crude cold water-soluble polysaccharides, crude boiling water-soluble polysaccharides , crude acid-soluble polysaccharides had strong scavenging hydroxyl radical and reducing power , the capacity increased with the increasing concentration of Smilax glabra polysaccharides; the scavenging hydroxyl radical and reducing power capacity of crude alkali-soluble polysaccharides is very poor. Conclusion Polysaccharides by using four kinds of different extraction methods have certain antioxidant activity, the antioxidant capacity to scavenge free radical in vitro showed positive correlation with the concentration of Smilax glabra polysaccharides, considering the yield of polysaccharides, EDTA?2Na aqueous solution extraction method is the best.
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Objective To investigate the protective effect of Smilax Glabra Roxb on the expression of vascular cell adhesion molecule-1 (VCAM-1) induced by IL-1 in human vascular endothelial cells (HUVECs). Methods Take human umbilical vascular endothelial cells as study subject, use immunofluorescence staining, serum pharmacology of the traditional Chinese medicine and flow cytometer. The level of VCAM-1 is represented by the mean fluorescence intensity. Results IL-1 increases the expression of VCAM-1, the expression of VCAM-1 in group of serum containing drugs is lower than control group. Conclusion Smilax Glabra Roxb can inhibit the effect of IL-1 induced increase of VCAM-1 expression in HUVECs.
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The ethanol extract of smilax glabra roxb (SG) (100mg/kg of body weight) reduced the blood level of glucose and plasma insulin levels of GK rat 4h after intraperitoneal administration (p< 0.01). While in the normal rat, it reduced only the glycemia. On isolated pancreas islets, SG concentration of 2mg – 4 mg/ml/h has no effect on insuline recretion. The finding suggests that hypoglycemic effect of SG is caused by the increase of the sensibility of target tissue to insuline.
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Rats , Blood Glucose , Diabetes Mellitus , Plant Proteins , Animal ExperimentationABSTRACT
Cryophylisat powder of ethanolic fluid extract of Smilax glabra (SG) was administered orally on mouse with a single dose of 1g/kg of body mass. Hypoglycemic effect exerted with 10.58% approximatively. Continous use of 7 days of this dose increased the hypoglycemic action. Oral SG effects manifested lately with less level than injectable SG. Apart from the increase of synthetization capacity of glycogene, SG promote the sensitivity of the tissues to insuline.
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Mice , Glucose Tolerance Test , Hypoglycemic AgentsABSTRACT
Cryophylisat powder of ethanolic fluid extract of Smilax glabra with the oral dose of 1.25g and 2.5g/kg of body mass (5-10 times of active dose on hypoglycemic effect was administered in rabbit in 15-30 days continously. These doses did not influenced on the body mass, the hematologies indices as well as the functional indices of lever and kidney. However, in microscopic image, there are some degenerative mainfestations of low levels of liver cells among two groups using the studied preparation.