Investigation of the possible protective effect of Smilax fluminensis steud. leaf in mice subjected to oxidative stress by paracetamol

Paracetamol (PCM) is a drug widely used by the population as an antipyretic and analgesic. If administered in high doses it can cause liver damage, leading to hepatoxicity. The genus Smilax, found in temperate and tropical regions, is traditionally used by the population through the extract of leaves and roots for several conditions, such as in the treatment of syphilis, diabetes, asthma and as a diuretic action. Through this, Smilax fluminensis leaf extracts were used to evaluate the protective effect against oxidative stress induced by a high dose of PCM in mice that received the drug and after receiving treatment with crude extract and fractions. Plasma analysis was performed using as partate aminotransferase (AST), alanine aminotransferase (ALT), glucose, triglycerides and cholesterol, in addition to biochemical techniques such as catalase (CAT), glutathione-S-transferase (GST), reduced glutathione (GSH), ascorbic acid (ASA), substances reactive to thiobarbituric acid (TBARS) and carbonylated proteins (CARBONYL) of liver, brain and kidneys. Fraction 1 of the extract was the most promising, decreasing the plasma levels of AST and ALT, the levels of CAT and GST of the liver, together with GSH and in the renal and brain tissue there was a decrease in carbonylated proteins (PCM + F1 versus PCM ). Besides, fraction 1 proved to be hypoglycemic and hypocholesterolemic. It is concluded that fraction 1 of Smilax fluminensis leaves has good antioxidant activity in the face of the damage caused by the high dose of paracetamol.

Ethyl acetate extract of Smilax glabra Roxb roots and its major active compound astilbin promote osteoblastogenesis in vitro by upregulating bone cell differentiation- associated genes

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.

Evaluation of anticancer effects of a pharmaceutically viable extract of a traditional polyherbal mixture against non-small-cell lung cancer cells / 中西医结合学报

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.

Study on Anti-tumor Effect of Ethanol Extract of Smilax trinervula and Its Different Polar Extract Parts / 中国药房

OBJECTIVE： To study the anti-tumor effect of ethanol extract of Smilax trinervula and its different polar extract parts， and to provide reference for the screening of anti-tumor active parts. METHODS： MTT method was used to detect the inhibitory rates of different concentrations of ethanol extract of S. trinervula， its petroleum ether， ethyl acetate， n-butanol and water layer extract parts to the proliferation of human hepatocellular carcinoma cell line HepG2， human lung cancer cell line A549， human breast cancer cell line MCF-7 and MDA-MB-231， human cervical cancer cell line HeLa and human ovarian cancer cell line HO-8910. Semi-inhibitory concentration （IC50） was calculated. A total of 80 KM mice were subcutaneously inoculated with S180 cell suspension in right forelimb axilla to induce tumor-bearing mice model. The mice were randomly divided into 8 groups： e.g. model group （normal saline， twice a day， i.g.）， cyclophosphamide （positive drug） group （0.025 g/kg， once a day， i.p.）， S. trinervula ethanol extract high-dose， medium-dose and low-dose groups， ethyl acetate part of ethanol extract high-dose， medium-dose and low-dose groups （0.1， 0.05， 0.025 g/kg by extract， twice a day， i.g.）， with 10 rats in each group. The mice in each group were given medicine for consecutive 14 days. The inhibition rate， spleen index and thymus index of mice in each group were measured after fasting for 12 hours after the last administration. RESULTS： In vitro experiments showed that S. trinervula and different polar extracts inhibited the proliferation of 6 kinds of tumor cells in significant dose-effect manner， especially ethyl acetate part of ethanol extract. IC50 of it to tumor cells was 40-210 μg/mL， that of it to MCF-7， MDA-MB-231 and HeLa cells were 70.56， 83.58， 44.67 μg/mL， respectively. In vivo experiments showed that the tumor mass of mice were decreased significantly in each administrations （P＜0.05 or P＜0.01）， the tumor mass of mice in S. trinervula ethanol extract high-dose group， ethyl acetate part of ethanol extract high-dose and medium dose groups were significantly lower than cyclophosphamide group （P＜0.01）. The spleen index of mice was decreased significantly in cyclophosphamide group， S. trinervula ethanol extract high-dose group， ethyl acetate part of ethanol extract high-dose and medium dose groups， compared with model group （P＜0.01）； thymus index of mice in cyclophosphamide group was increased significantly， compared with model group and other administration groups （P＜0.01）. CONCLUSIONS： The ethyl acetate part of ethanol extract of S. trinervula has the best anti-tumor effect and less immunosuppressive effect.

Mechanism of Smilax glabra in treatment of gout based on network pharmacology / 中草药

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.

Spectrum-effect relationship between UPLC fingerprint of Smilax china and anti-pelvic inflammation in rats / 中国中药杂志

To study the correlation between ultra high performance liquid chromatography( UPLC) fingerprint of Smilax china and its anti-pelvic inflammatory effect,and to explore the pharmacodynamic material basis of S. china against pelvic inflammatory disease.UPLC fingerprints of 10 batches of S. china from different habitats were established,and the values of SOD,MDA,TNF-α,and IL-6 in rats with pelvic inflammation were measured. The weight of each single pharmacodynamics index to the total efficacy was determined by analytic hierarchy process,and the contribution of each peak in fingerprints to the each single pharmacodynamics index and total efficacy was analyzed by the grey relational analysis. Then the structures of chemical constituents at the identified peaks were confirmed by comparing with the reference substance. The 27 common characteristic peaks of UPLC fingerprints were all related to the anti-pelvic inflammation effect of S. china,of which 13 peaks were identified as peak 2( 3,5-dihydroxy-2-methylbenzoic acid-3-O-glucoside),peak 3( chlorogenic acid),peak 5( 2,7,4-trihydroxydihydroflavone-5-O-glucoside),peak 6( 7,4-dihydroxydihydroflavonol-5-O-glucoside),peak 7( taxifolin-7-O-glucoside),peak 9( taxifolin),peak 10( polydatin),peak 11( oxyresveratrol),peak 12( astilbin),peak15( resveratrol),peak 16( quercitrin),peak 18( engeletin) and peak 24( kaempferol). The correlation degree of 21 peaks and the total efficacy was greater than 0. 8,and the top 10 ranked by correlation degree were as follows: peak 1,3,7,19,18,17,4,11,16,and 21. The results showed that the anti-pelvic inflammation effect of S. china was achieved by the combined action of pharmacodynamic substances. In order to control the quality of S. china and its prepared slices more effectively,the index components of content detection should be selected reasonably.

Content changes regularity of flavonoids in Smilax glabra by QAMS method / 中草药

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.

Analysis of Volatile Components in Smilax Glabra Robx. and Its Adulterant / 中国药师

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.

Two new compounds from Smilax glaucochina / 中国中药杂志

A new sesquiterpenes named glaucochinarol A (1) and a new phenylpropane glycoside named glcacochinaside A (2), together with six known ones, including trichothecolone (3), β-D-(6-O-trans-feruloyl)fructofuranosyl-α-D-O-glucopyranoisde (4), 3,4,5-trimethoxyphenyl-β-D-glucopyranoside (5), (4R)-p-menth-1-ene-7,8-diol-7-O-β-D-glucopyranoside (6), naringenin (7), and emodin-8-O-β-glucoside (8) were isolated from smilax glaucochina warb. Their structures were elucidated on the basis of NMR, MS and published data. Compounds 3-8 were isolated from the species for this first time.

High-Performance Liquid Chromatography Fingerprint Analysis of Smilacis Glabrae Rhizoma and Its Adulterants / 中国药学杂志

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.

Chemical constituents from rhizome of Smilax davidiana / 中草药

Objective: To study on the chemical constituents from the rhizome of Smilax davidiana. Methods: The compounds were separated and purified by sephadex LH-20 column chromatography and high performance liquid chromatography. Their structures were identified by spectroscopic analysis and comparison with literatures. Results: Twenty compounds were isolated and identified as aromadendrin7-O-β-D-glucopyranoside (1), dalbergin (2), 3,5,7,4′-tetrahydroxyflavanone-7-O-β-D-glucopyranoside (3), kaempferol-7-O-β-D-glucopyranoside (4), quercetin-7-O-β-D-glucopyranoside (5), quercetin-3-O-β-L-rhamnopyranoside (6), 4,6- dihydroxy-2-O-(β-D-glucopyranosyl)-acetophenone (7), 3,5-dihydroxy-4-O-(β-D-glucopyranosyl) acetophenone (8), 2,4,6- trihydroxy-acetophenone-4-O-β-D-glucopyranoside (9), 2,4,6-trihydroxylacetophenone-2,4-di-O-β-D-glucopyranoside (10), epicatechin (11), latifolin (12), 3′-O-(E-4-coumaroyl)-quinic acid (13), 5-O-caffeoylquinic acid butyl ester (14), 5,5′- dimethoxylariciresinol 4′-O-β-D-glucopyranoside (15), 1-cerotoylglycerol (16), cinchonain Ib (17), adenosine (18), resveratrol (19), and 3,4,5-trimethoxyphenyl-1-β-D-glucopyranoside (20). Conclusion: Compounds 4—9, and 12—16 are isolated from Smilax genus for the first time, and compounds 1—3, 17, and 18 are from S. davidiana for the first time.

Chemical constituents from the rhizomas of Smilax glauco-china / 中成药

AIM To study the chemical constituents from the rhizomas of Smilax glauco-china Warb.METHODS The n-butanol fraction of ethanol extract of S.glauco-china was isolated and purified by silica,Sephadex LH-20 and semi-preparative column,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Ten compounds were isolated and identified as phenethanol-β-D-gentiobioside (1),2-phenylethyl-O-β-D-xylopyranosyl-(1 →6)-β-D-glucopyranoside (2),phenylethyl D-rutinoside (3),phenylethyl β-D-glucoside (4),hydrangeifolin Ⅰ (5),icariside D1 (6),calophymembranside B (7),2-hydroxyphenol-1-O-β-D-glucopyranosyl-(6 → 1)-α-L-rhamnopyranoside (8),β-sitosterol (9),daucosterol (10).CONCLUSION All the compounds are isolated from this plant for the first time.

Studies on chemical constituents of rhizomes of Smilax trinervula / 中国中药杂志

The chemical constituents were separated and purified from the 70% ethanol extract of Smilax trinervulaby various chromatographic methods including silica gel, Sephadex LH-20, MCI and preparative HPLC. Their structures were obtained and identified by analysis of the spectroscopic data. Compounds 1-11 were separated from this genus for the first time. Compound 12 was obtained from S. trinervula for the first time.

Evaluations of cytotoxicity of Smilax myosotiflora and its effects on sexual hormone levels and testicular histology in male rats

Objective: To investigate the cytotoxicity of Smilax myosotiflora (S. myosotiflora) methanolic extract and its effects on sexual hormone levels and testicular histology in male rats. Methods: The cytotoxicity of S. myosotiflora methanolic extract was investigated by employing brine shrimp lethality assay. Forty eight male rats were randomly divided into four groups (Groups I-IV) of 12 each. Rats in Group I were administered with 0.5 mL of distilled water (vehicle), whilst Groups II, III and IV received 200, 400 and 800 mg/kg of the methanolic extract of S. myosotiflora in 0.5 mL of the vehicle, respectively. Male rats treated with continuous daily dosing were killed and necropsied after a total dose period of 60 days. Sexual hormones were assayed and histological examination of testes was performed according to standard methods. Results: S. myosotiflora extracts did not produce any cytotoxicity to brine shrimp in all concentrations tested. Serum testosterone level was significantly higher in rats treated with high dose of S. myosotiflora. Testicular histology showed normal architecture with all stages of spermatogenesis in all experimental groups. Conclusions: The present work confirmed that S. myosotiflora extract improves reproductive functions, without any cytotoxic activity and produces no histological changes to the testes.

Axillary bud and pericycle involved in the thickening process of the rhizophore nodes in Smilax species / Gema axilar e periciclo envolvidos no processo de espessamento nodal do rizóforo em espécies de Smilax

AbstractThe species of the genus Smilax, popularly known as sarsaparilla, are widely used in folk medicine due to the antirheumatic properties of its underground structures. Smilax fluminensis and S. syphilitica occur in forested areas and form thickened stems called rhizophores from which adventitious roots grow. To provide information for more accurate identification of the commercialised product and for elucidating the process of stem thickening, a morphology and anatomy study of the underground organs of the two species was conducted. The adventitious roots differ in colour and diameter depending on the stage of development. They are white and have a larger diameter in the early stages of development, but as they grow, the adventitious roots become brown and have a smaller diameter due to the disintegration of the epidermis and virtually the entire cortex. In brown roots, the covering function is then performed by the lignified endodermis and the remaining walls of the cells from the last parenchyma cortical layer. These results are similar to those found in studies of other Smilax and suggest that the anatomy of the roots can be useful for identifying fraud in commercialised materials. The thickening process of the nodal regions of the rhizophores in both species involves the activity of axillary buds and pericyclic layers.

ResumoAs espécies de Smilax, conhecidas popularmente como salsaparrilha, são amplamente utilizadas na medicina tradicional devido às propriedades antirreumáticas das estruturas subterrâneas. Smilax fluminensis e S. syphilitica ocorrem em áreas florestais e formam caules espessados denominados rizóforos a partir dos quais são emitidas raízes adventícias. Com o intuito de fornecer informações para a identificação mais precisa do material comercializado e no entendimento do processo de espessamento do caule, foi realizado o estudo morfológico e anatômico dos órgãos subterrâneos das duas espécies. As raízes adventícias apresentam diferenças na coloração e no diâmetro dependendo da fase de desenvolvimento. As raízes nas fases iniciais do desenvolvimento são brancas e possuem diâmetro maior, porém com o desenvolvimento, devido à desintegração da epiderme e de praticamente todo o córtex, as raízes tornam-se marrons e de diâmetro menor. Nas raízes marrons, a função de revestimento passa a ser exercida pela endoderme lignificada e pelas paredes remanescentes das células da penúltima camada cortical. Os resultados são semelhantes aos encontrados nos estudos de outras Smilax e sugerem que a anatomia das raízes pode ser útil na identificação de fraudes em materiais comercializados. O processo de espessamento das regiões nodais dos rizóforos nas duas espécies envolve a atividade das gemas axilares e de camadas pericíclicas.

Smilax china L. rhizome extract inhibits nuclear factor-κB and induces apoptosis in ovarian cancer cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the antitumor effects and associated mechanisms of extract of the Smilax china L. rhizome (SCR) on ovarian cancer cells.</p><p><b>METHODS</b>Ovarian cancer cells A2780 were treated with different concentrations of SCR extract (SCRE), and compared with controls. Effects on cell growth were evaluated by cell counting kit-8 (CCK-8) assay; proliferation effects by EdU incorporation assay; cell cycle by propidium iodide staining; apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide; cellular distribution of nuclear factor-κB (NF-κB) by immunofluorescence; protein levels of NF-κB, caspase-3, poly-adenosine diphosphate (ADP)-ribose polymerase (PARP), Bcl-2-associated X protein (Bax), cellular inhibitor of apoptosis (cIAP)-1, anti-X-linked inhibitor of apoptosis protein (XIAP), B-cell lymphoma-extra large (Bcl-XL), B-cell lymphoma-2 (Bcl-2) and AKT by Western blotting; and effects of SCRE combined with cisplatin or adriamycin on A2780 cells by CCK-8 assay.</p><p><b>RESULTS</b>SCRE suppressed A2780 cell proliferation in a dose-dependent manner (P<0.05,P<0.01), arrested cells in G2/M phase and induced apoptosis by activating caspase-3, PARP and Bax. SCRE treatment also correlated with inhibition of NF-κB and downregulation of Bcl-2, Bcl-XL, cIAP-1, XIAP and AKT. SCRE can promote chemosensitivity to cisplatin and adriamycin in A2780 cells (P<0.01).</p><p><b>CONCLUSION</b>SCR effectively inhibits NF-κB, induces apoptosis and reduces chemoresistance to cisplatin and adriamycin in ovarian cancer cells, which might be its molecular basis for treating ovarian cancer.</p>

Antiobesity effects of the water-soluble fraction of the ethanol extract of Smilax china L. leaf in 3T3-L1 adipocytes

BACKGROUND/OBJECTIVES: Several medicinal properties of Smilax china L. have been studied including antioxidant, anti-inflammatory, and anti-cancer effects. However, the antiobesity activity and mechanism by which the water-soluble fraction of this plant mediates its effects are not clear. In the present study, we investigated the lipolytic actions of the water-soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) in 3T3-L1 adipocytes. MATERIALS/METHODS: The wsSCLE was identified by measuring the total polyphenol and flavonoid content. The wsSCLE was evaluated for its effects on cell viability, lipid accumulation, glycerol, and cyclic adenosine monophosphate (cAMP) contents. In addition, western blot analysis was used to evaluate the effects on protein kinase A (PKA), PKA substrates (PKAs), and hormone-sensitive lipase (HSL). For the lipid accumulation assay, 3T3-L1 adipocytes were treated with different doses of wsSCLE for 9 days starting 2 days post-confluence. In other cell experiments, mature 3T3-L1 adipocytes were treated for 24 h with wsSCLE. RESULTS: Results showed that treatment with wsSCLE at 0.05, 0.1, and 0.25 mg/mL had no effect on cell morphology and viability. Without evidence of toxicity, wsSCLE treatment decreased lipid accumulation compared with the untreated adipocyte controls as shown by the lower absorbance of Oil Red O stain. The wsSCLE significantly induced glycerol release and cAMP production in mature 3T3-L1 cells. Furthermore, protein levels of phosphorylated PKA, PKAs, and HSL significantly increased following wsSCLE treatment. CONCLUSION: These results demonstrate that the potential antiobesity activity of wsSCLE is at least in part due to the stimulation of cAMP-PKA-HSL signaling. In addition, the wsSCLE-stimulated lipolysis induced by the signaling is mediated via activation of the beta-adrenergic receptor.

Effects of different extraction methods on yield of Smilax glabra polysaccharides and antioxidant activity / 国际中医中药杂志

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.

Screening for the Bioactive Fraction of Smilax china L. on Inhibition of Experimentally Induced Benign Prostatic Hyperplasia / 医药导报

Objective To study the effect of extracts of Smilax china L. on inhibition the experimentally induced benign prostatic hyperplasia ( BPH) , and screen the effective fraction. Methods The BPH model was built on the castrated rats by subcutaneous injection of testosterone propionate. Male rats were randomly divided into eight groups ( n=6 ):sham operation, model control, petroleum ether fraction, acetic ether fraction, n-butyl alcohol fraction, water fraction, macroporous resin fraction ( FMR) , and total extracts group. The rats were treated with testosterone propionate by subcutaneous injection for consecutive 3 weeks. Meanwhile, rats were orally administrated with the six extract fractions of S. china L. After the last administration, serum was separated for the determination of prostatic acid phosphatase ( PACP ) , prostate was weighed and histopathological examination was carried out to evaluate the inhibitory effect of S. china L. against BPH. Results All of the six fractions from S. china L. could inhibit BPH, and the n-butanol fraction, water fraction and FMR showed better inhibitory effect, which significantly decreased the prostatic index by 52. 80%, 50. 93% and 67. 70%, respectively, remarkably reduced serum PACP, and notably improved the prostate gland morphology compared with the model group. Among the three fractions, FMR showed the strongest effect against BPH. Conclusion S. china L. ameliorates the experimentally prostatic hyperplasia, and FMR showes the best effect, which might be the bioactive components against BPH.

Effect of Different Extracts of Smilax China L. on the Activity of Ovarian Cancer Cells / 中国药师

Objective: To investigate the effects of different extracts of Smilax china L on the activity of ovarian cancer cells. Methods:Solvent extraction method was used to extract the active ingredients of Smilax china L. , and CCK-8 assay method was ap-plied to detect the influence of different Smilax china L. extracts (0, 50, 100, 150 and 200μg·ml-1 ) on the survival rate of ovarian cancer cells including low invasiveness A2780 cells and high invasiveness HO-8910PM cells. At the same time, the status of the two kinds of ovarian cancer cells at different time points (24, 48 and 72 h) was observed. Results:The IC50 of N-butanol extracts (SCR-B) on HO-8910 and A2780 ovarian cancer cells was 47. 5 μg· ml-1 and 69. 2 μg· ml-1 , respectively, and that of ethyl acetate ex-tracts (SCR-E) on A2780 and HO-8910 cells was 147. 9 μg· ml-1 and 166. 0 μg· ml-1, respectively. Smilax china L. extracts had the inhibition against both A278 and HO-8910PM ovarian cells in a dose-and time-dependent manner. Conclusion:The inhibitory activity of SCR-B against ovarian cancer cells is stronger than that of SCR-E, and SCR-B has stronger inhibition against A2780 cells than against HO-8910 cells. SCR-B has better inhibition against ovarian cancer cells.