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.

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.

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.

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.

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.

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>

Study of the mechanisms underlying increased glucose absorption in Smilax china L. leaf extract-treated HepG2 cells / 한국영양학회지

PURPOSE: Previous studies have shown that treatment with Smilax china L. leaf extract (SCLE) produces antidiabetic effects due to alpha-glucosidase inhibition. In this study, we examined the mechanism underlying these antidiabetic effects by examining glucose uptake in HepG2 cells cultured with SCLE. METHODS: Glucose uptake and glucokinase activity were examined using an assay kit. Expression of glucose transporter (GLUT)-2, GLUT-4, and HNF-1alpha was measured by RT-PCR or western blot. RESULTS: Treatment with SCLE resulted in enhanced glucose uptake in HepG2 cells, and this effect was especially pronounced when cells were cultured in an insulin-free medium. SCLE induced an increase in expression of GLUT-2 but not GLUT-4. The increase in the levels of HNF-1alpha, a GLUT-2 transcription factor, in total protein extract and nuclear fraction suggest that the effects of SCLE may occur at the level of GLUT-2 transcription. In addition, by measuring the change in glucokinase activity following SCLE treatment, we confirmed that SCLE stimulates glucose utilization by direct activation of this enzyme. CONCLUSION: These results demonstrate that the potential antidiabetic activity of SCLE is due at least in part to stimulation of glucose uptake and an increase in glucokinase activity, and that SCLE-stimulated glucose uptake is mediated through enhancement of GLUT-2 expression by inducing expression of its transcription factor, HNF-1alpha.

Screening of the Different Concentrations of Ethanol Extracts from Smilax China for Its Antiinflammatory Activity / 医药导报

Objective To study the effects of the different concentrations of ethanol extracts of Smilax China on ear edema in mice and granuloma in rats,and to provide an evidence for optimizing the extraction process. Methods Effects of different concentrations of ethanol extracts of Smilax China on the xylene-induced ear edema in mice and the cotton ball-induced granuloma hyperplasia in rats were tested . Results Compared with the model controls,70% ethanol extracts of Smilax China at high,medium and low doses significantly inhibited ear edema in mice (t=2. 58,P<0. 05;t=2. 28,P<0. 05;t=2. 17,P<0. 05) and reduced the granuloma hyperplasia in rats(t=5. 28,P<0. 01;t=5. 24,P<0. 01;t=5. 17,P<0. 01). Conclusion The 70% ethanol extracts of Smilax China at three doses present the most active antiinflammatory effect,confirmed in both mice ear edema and rats granuloma models.

Study on antioxidative, antidiabetic and antiobesity activity of solvent fractions of smilax china L. leaf extract / 한국영양학회지

Smilax china L., a native plant found in Asian countries, has several medicinal properties including antioxidant, anti-inflammatory, and anti-cancer effects. Although the root of the plant is commonly used as traditional herbal medicine in Korea and China, the medicinal properties of the leaves have not gained the same attention. In this study, we analyzed the antioxidant activity, alpha-glucosidase inhibitory effect and lipid accumulation inhibition effect of Smilax china L. leaf water extract (SCLE) and its solvent fractions. SCLE was fractionated by using a series of organic solvents, including ethylacetate (EA) and n-butanol (BuOH). The EA fraction had the highest total polyphenol content (440.20 +/- 12.67 mg GAE/g) and total flavonoid content (215.14 +/- 24.83 mg QE/g). The radical scavenging activity IC50 values of the EA fraction for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) were 0.022 mg/mL and 0.13 mg/mL, respectively. Further, SOD-like activity and reducing power values of the EA fraction were higher than those of the other fractions. However, both the alpha-glucosidase and lipid accumulation inhibition assays showed that the BuOH fraction (83.35 +/- 4.18% at 1 mg/mL) and water extract (11.27 +/- 2.67%) were more effective than the EA fraction (64.13 +/- 6.35%, and 45.66 +/- 7.20%). These results provide new insights into the potential anti-diabetic and anti-obesity effects of Smilax china L. leaf.

Free radical scavenging and antioxidant enzyme fortifying activities of extracts from Smilax china root

The extract from Smilax china root has been used as medicinal remedy and reported to retain antimicrobial and antimutagenic acitivities. In this study, a possible presence of antioxidant activity of Smilax china root extract was investigated. Methanol extract (Me) revealed the presence of high 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity (IC50 7.4 microg/ml) and protective property of cell's viability. Further fractionation with various solvent extraction and assay showed high levels of DPPH free radical scavenging activity in the ethyl acetate, butanol and water extracted fractions. In addition, V79-4 cells treated with Me of Smilax china root induced an increase of superoxide dismutase, catalase and glutathione peroxidase activities in a dose-dependent manner between 4-100 microg/ml. These results suggest that the medicinal component of the root of Smilax china extracts also contains antioxidant activity.

Free radical scavenging and antioxidant enzyme fortifying activities of extracts from Smilax china root

The extract from Smilax china root has been used as medicinal remedy and reported to retain antimicrobial and antimutagenic acitivities. In this study, a possible presence of antioxidant activity of Smilax china root extract was investigated. Methanol extract (Me) revealed the presence of high 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity (IC50 7.4 microg/ml) and protective property of cell's viability. Further fractionation with various solvent extraction and assay showed high levels of DPPH free radical scavenging activity in the ethyl acetate, butanol and water extracted fractions. In addition, V79-4 cells treated with Me of Smilax china root induced an increase of superoxide dismutase, catalase and glutathione peroxidase activities in a dose-dependent manner between 4-100 microg/ml. These results suggest that the medicinal component of the root of Smilax china extracts also contains antioxidant activity.

Flavonoids and stilbenes from Smilax china / 中草药

Objective To study the chemical constituents of Smilax china.Methods The compounds were isolated by silica gel,Sephadex LH-20,ODS chromatography,and semi-preparative HPLC.Their structures were elucidated on the basis of MS and NMR spectroscopic analyses.Results Seven flavonoids and four stilbenes were isolated and identified as dihydrokaempferol-5-O-?-D-glucoside(Ⅰ),engeletin (Ⅱ),isoengeletin(Ⅲ),dihydroquercetin-3-O-glycoside(Ⅳ),3,5,7,3',5'-pentahydroxy-flavanonol (Ⅴ),astilbin(Ⅵ),quercetin-3'-Oglycoside(Ⅶ),piceid(Ⅷ),scirpusin A(Ⅸ),resveratrol(Ⅹ),and oxyresveratrol(Ⅺ).Conclusion CompoundsⅣ—Ⅸare isolated from this plant for the first time,among which the ~(13)C-NMR data of compoundⅨis reported firstly.

Study on Antioxidant Stilbenes from the Rhizomes of Smilax china / 中药新药与临床药理

Objective To study the stilbene constituents from the traditional Chinese medicine Smilax china and to determine their antioxidant activity. Methods The compounds were separated and purified by column chromatography with silica gel, RP C18, and Sephadex LH- 20, and were identified by IR, MS, NMR. DPPH method was used to evaluate the free radical scavenging activity of the isolated compounds. Results Three compounds were isolated from the EtOAc fraction of the rhizomes of S. china and were identified as: resveratrol (1), oxyresveratrol (2) and 3, 5, 3′ , 4′ - tetrahydroxylstilbene (3). Compounds 1～ 3 showed strong antioxidant activity, and could scavenge DPPH free radicals, effectively. At the concentration of 50 ? mol/L, their DPPH free radical scavenging rates were 79.47 % , 89.89 % and 93.86 % , respectively. Conclusion Stilbenes might be the material foundation of antioxidant activities of rhizomes of S. china.