Anti-Adipogenic Activity of Secondary Metabolites Isolated from Smilax sieboldii Miq. on 3T3-L1 Adipocytes.

Smilax sieboldii, a climbing tree belonging to Smilacaceae, has been used in traditional oriental medicine for treating arthritis, tumors, leprosy, psoriasis, and lumbago. To evaluate the anti-obesity effects of S. sieboldii (Smilacaceae), we screened methylene chloride (CH2Cl2), ethyl acetate (EtOAc), aqueous-saturated n-butanol, and ethanol (EtOH) extracts of the whole plant at various concentrations to inhibit adipogenesis in adipocytes. The 3T3-L1 cell line with Oil red O staining with the help of fluorometry was used as an indicator of anti-obesity activity. Bioactivity-guided fractionation of the EtOH extract and subsequent phytochemical investigation of the active CH2Cl2- and EtOAc-soluble fractions resulted in the isolation of 19 secondary metabolites (1-19), including a new α-hydroxy acid derivative (16) and two new lanostane-type triterpenoids (17 and 18). The structures of these compounds were characterized using various spectroscopic methods. All the isolated compounds were screened for adipogenesis inhibition at a concentration of 100 µM. Of these, compounds 1, 2, 4-9, 15, and 19 significantly reduced fat accumulation in 3T3-L1 adipocytes, especially compounds 4, 7, 9, and 19, showing 37.05 ± 0.95, 8.60 ± 0.41 15.82 ± 1.23, and 17.73 ± 1.28% lipid content, respectively, at a concentration of 100 µM. These findings provide experimental evidence that isolates from S. sieboldii extracts exert beneficial effects regarding the regulation of adipocyte differentiation.

Chemical profile, antioxidant potential and toxicity of Smilax brasiliensis Sprengel (Smilacaceae) stems.

Smilax brasiliensis Sprengel is a monocotyledon of the Smilacaceae family, native to the Brazilian Cerrado, popularly known as "salsaparrilha" or "japecanga". In this study, the ethanol extract (EE) and the hexane (HEXF), dichloromethane (DCMF), ethyl acetate (ACF), and hydroethanol (HEF) fractions of the stems were obtained. The chemical composition was determined, the contents of phenolic compounds and flavonoids were quantified, and the antioxidant potential and the cytotoxic effect on Artemia salina were evaluated. Fatty acid esters, hydrocarbons, and phytosterols were identified in the HEXF analyzed by gas chromatography - mass spectrometry (GC-MS). The EE and DCMF, ACF, and HEF were analyzed by liquid chromatography coupled to a diode array detector and mass spectrometer (LC-DAD-MS), and the identified constituents included glycosylated (rutin, 3-O-ß-galactopyranosyl quercetin, 3-O-ß-glucopyranosyl quercetin, O-deoxyhexosyl-hexosyl quercetin, O-deoxyhexosyl-hexosyl kaempferol, O-deoxyhexosyl-hexosyl O-methyl quercetin, and others), and non-glycosylated (quercetin) flavonoids, phenylpropanoids (3-O-E-caffeoyl quinic acid, 5-O-E-caffeoyl quinic acid, O-caffeoyl shikimic acid, and others), neolignan, steroidal saponin (dioscin), and N-feruloyltyramine. The EE, DCMF, and ACF showed high total contents of phenolic compounds (112.99, 175.71, and 524.02 µg of GAE/mg, respectively), and in the ACF and DCMF a great content of flavonoids was also quantified (50.08 and 31.49 µg of QE/mg, respectively). The EE, DCMF, ACF, and HEF exhibited great antioxidant potential by DPPH (IC50 1.71 - 32.83 µg/mL) and FRAP (IC50 0.63 - 6,71 µg/mL) assays. A maximum cytotoxic activity on A. salina of 60% was observed for the DCMF (LC50 = 856.17 µg/mL). This study contributes to the phytochemical study of S. brasiliensis since these compounds were identified for the first time in the stems of this species. The S. brasiliensis stems demonstrated to be a rich source of polyphenols compounds and exhibited high antioxidant potential without toxicity. Thus, extract and fractions obtained from the S. brasiliensis stems can be used in food supplements or as natural antioxidants in the food industry.

Identification of the Constituents of Ethyl Acetate Fraction from Smilax china L. and Determination of Xanthine Oxidase Inhibitory Properties.

The aim of this work was to investigate the xanthine oxidase (XO)-inhibitory activity of ethanol extracts from Smilax china L. and to identify the active compounds in the ethyl acetate (EtOAc) fraction. Extraction of ethanol extracts from Smilax china L. and then ethanol extracts were concentrated, and the polyphenolic compounds were extracted with petroleum ether (PE), chloroform, EtOAc, n-butanol (n-BuOH), and residual ethanol fractions. Their effects on XO activity were then compared separately. The polyphenolic components of the EtOAc fraction were identified by HPLC and HPLC-mass spectrometry (HPLC-MS) analysis. Kinetic analysis demonstrated that all these extracts showed XO-inhibitory properties, and among them the EtOAc fraction had the strongest inhibitory effect (IC50 = 101.04 µg/mL). The inhibitory constant (Ki) of the EtOAc fraction on XO activity was 65.20 µg/mL, showing excellent inhibition on XO in the competitive mode. Sixteen compounds were identified from the EtOAc fraction. The study demonstrates that the EtOAc fraction of Smilax china L. may be a potential functional food to inhibit XO activity.

Structural characterization and anti-inflammatory activity of a novel neutral polysaccharide isolated from Smilax glabra Roxb.

Crude polysaccharides isolated from Smilax glabra were screened for anti-inflammatory activity using mice ear swelling animal experiments, during which the neutral polysaccharide S1 was identified. The structural characteristics and anti-inflammatory effects of the anti-inflammatory S1 polysaccharide were then investigated. The results showed that S1 was mainly composed of rhamnose, arabinose, galactose, glucose, xylose, and mannose. The structure of the main chain consisted of →6)-α-Galp-(1 â†’ 6)-ß-Galp-(1 â†’ 4)-α-Xylp-(1 â†’ 6)-ß-Galp-(1→, with branched chains comprising α-Araf-(1 â†’ 4)-α â†’ Manp-(1 â†’ and ß-Rhap-(1 â†’ 4)-α-Glcp-(1 â†’ units. Furthermore, S1 did not have a triple helix conformation. S1 could inhibit NO secretion, reduce the levels of pro-inflammatory factors (IL-6 and TNF-α), and significantly reduce LPS-stimulated inflammatory damage in RAW 264.7 cells by inhibiting activation of the NF-κB (p65) pathway. These results shed light on the possibility of S1 to be developed as a novel anti-inflammatory drug for therapeutic purposes.

A comprehensive quality control evaluation for standard decoction of Smilax glabra Roxb based on HPLC-MS-UV/CAD methods combined with chemometrics analysis and network pharmacology.

An effective, sensitive, and rapid method was developed for the quality control evaluation of the standard decoction of Smilax glabra Roxb (SGR). SGR is a primary ingredient of the traditional functional foods of turtle jelly and SGR tea. Chemometrics, Network Pharmacology, and molecular docking were used to screen for six quality markers. Multiple extraction parameters were optimized. HPLC-UV/CAD-QAMS was used to rapidly quantify the six quality markers (neoastilbin, astilbin, neoisoastilbin, isoastilbin, quercitrin, and isoengeletin) in 10 batches of the standard decoction of SGR samples. The relative correction factor (RCF) values of the five compounds were close to 1, demonstrating that the charged aerosol detection (CAD) showed a consistent response to compounds with similar parent nucleus structures. This method can serve as a guide for rapid quantitative analysis of the multi-components of the SGR standard decoction and all the traditional functional foods of turtle jelly with the homology of medicine.

Antimicrobial Activity of Smilax china L. Root Extracts against the Acne-Causing Bacterium, Cutibacterium acnes, and Its Active Compounds.

The root of Smilax china L. is used in traditional Korean medicine. We found that the Smilax china L. root extract has strong antimicrobial activity against two Cutibacterium acnes strains (KCTC 3314 and KCTC 3320). The aim of this study was to identify the beneficial properties of Smilax china L. extracts for their potential use as active ingredients in cosmetics for the treatment of human skin acne. The high-performance liquid chromatography (HPLC) and liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC/QTOF/MS) methods were used to obtain the profile of secondary metabolites from the ethyl acetate-soluble fraction of the crude extract. Agar diffusion and resazurin-based broth microdilution assays were used to evaluate antimicrobial activity and minimum inhibitory concentrations (MIC), respectively. Among the 24 metabolites, quercetin, resveratrol, and oxyresveratrol were the most potent compounds against Cutibacterium acnes. Minimum inhibitory concentrations of quercetin, resveratrol, and oxyresveratrol were 31.25, 125, and 250 µg/mL, respectively.

Smilax glabra Roxb.: A Review of Its Traditional Usages, Phytochemical Constituents, Pharmacological Properties, and Clinical Applications.

Smilax glabra Roxb. (SGB) is a medicinal plant widely distributed in 17 countries worldwide. It is the primary raw material of the world-famous and best-selling functional food and beneficial tea. SGB was first recorded in Ben Cao Jing Ji Zhu of the Southern and Northern Dynasties (420-589 AD) and was reported for nutritional and medicinal properties for thousands of years. This review searched PubMed, Web of Science, and other databases for relevant literature on SGB species until April 2022. It aims to provide more integrated thinking, detailed awareness, and better knowledge of SGB. More than 200 chemical components have been discovered, including flavonoids, phenolic, phenolic acids, stilbenes, organic acids, phenylpropanoids, and others. Previous studies have demonstrated that SGB and its active ingredients show a wide range of pharmacological effects, including anti-infective, anti-cancer, anti-inflammatory, antioxidant, cardiovascular protection, etc. However, many studies on the biological activity of this plant were mainly based on crude extracts and active ingredients, and there is a lack of clinical studies and toxicity studies to support the development of drug design, development, and therapy. In summary, this review will provide specific and valuable suggestions and guidelines for further research and application of this plant in the medicinal field.

Combining network pharmacology, RNA-seq, and metabolomics strategies to reveal the mechanism of Cimicifugae Rhizoma - Smilax glabra Roxb herb pair for the treatment of psoriasis.

BACKGROUND: Psoriasis is a prevalent chronic inflammatory skin condition marked by immune cell infiltration and keratinocyte abnormal proliferation. Cimicifugae Rhizoma - Smilax glabra Roxb (CS) herb pair, the main component of Shengma Detoxification Decoction, has been proven effective for the treatment of psoriasis. However, the mechanism is yet to be deciphered. PURPOSE: To explore the mechanism of CS for the treatment of psoriasis. METHODS: The imiquimod-induced psoriasis-like lesion mouse model was used to identify the targets and the molecular mechanisms of CS. Network pharmacology combined with RNA-seq strategy was employed to predict the targets and mechanisms of CS for psoriasis. Metabolomics approaches were used to demonstrate the complexity of CS for the treatment of psoriasis. Finally, a compound-response-enzyme-gene network was constructed based on the multi-omics results to elucidate potential connections. RESULTS: The CS herb pair could significantly improve psoriatic lesions and reduce the inflammatory cell infiltration and proliferation of keratinocytes in skin lesions. Network pharmacology predicted that TNF, JNK, IL-6, and IL-1ß could be potential targets. RNA-seq data revealed that CS could significantly regulate genes and signaling pathways associated with Th17 responses, such as IL-36, IL-1ß, CCl2, CXCL16, keratin 14, keratin 5, and antimicrobial peptides S100A8 and S100A9 well as MAPK, mTOR, and other signaling pathways. Further experimental data validated that CS treatment remarkably reduced the expression of inflammatory cytokines and factors, such as CCL2, CCL7, IL1F6, IL-17, IL-23, IL-1ß, TNF-α, and IL-6, and inhibited the phosphorylation of p38 and ERK1/2. This indicated that CS exerts its therapeutic effect by inhibiting the MAPK signaling pathways. In addition, metabolomic analyses demonstrated that CS treatment improved seven metabolic pathways, these included phenylalanine, tyrosine, pyruvate metabolism, carnitine metabolism, etc. Four key metabolites (L-Arginine, L-Phenylalanine, L-Carnitine, O-Acetylcarnitine) and nine differential genes (CMA1, PCBD2, TPSAB1, TPSB2, etc.) were identified that affected amino acid metabolism, carnitine metabolism, and other pathways contributing to the infiltration of Th17 cells in psoriatic lesions. CONCLUSION: CS could alleviate IMQ-induced psoriasis-like dermatitis by reducing the expression of cytokines and chemokines mediated by the MAPK pathway, and improved amino acid and carnitine metabolism in vivo. Our study is the first to demonstrate the complex mechanism of CS for the treatment of psoriasis and provides a new paradigm to elucidate the pharmacological effects of Traditional Chinese Medicine (TCM) drugs for psoriasis from multiple perspectives.

Quercetin, Engelitin and Caffeic Acid of Smilax china L. Polyphenols, Stimulate 3T3-L1 Adipocytes to Brown-like Adipocytes Via ß3-AR/AMPK Signaling Pathway.

The aim of the present study was to investigate the browning effects mechanism of Smilax china L. polyphenols (SCLP) and its monomer. In this study, polyphenols (SCLP, engeletin, quercetin and caffeic acid) markedly suppressed lipid accumulation. Polyphenols significantly up-graded the expression of protein kinase A (PKA), adipose triglyceride lipase (ATGL), peroxisome proliferators-activated receptors alpha (PPARα), carnitine palmitoyl transferase (CPT) and acyl-CoA oxidase (ACO) to promote lipolysis and ß-oxidation. Moreover, polyphenols greatly enhanced mitochondrial biogenesis in adipocytes, as demonstrated by the expression of Nrf1 and Tfam were up-regulated. Furthermore, polyphenols treatment greatly up-regulated the browning program in adipocytes by increased brown-specific genes and proteins uncoupling protein 1 (UCP-1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) and PR domain containing 16 (PRDM16), as well as beige-specific genes (Tmem26, Tbx1, CD137, Cited1), especially engeletin. Further research found that the brown-specific markers were decreased by antagonist treatment of AMPK or ß3-AR, but polyphenols treatment reversed the effect of antagonists and improved the expression of UCP-1, PRDM16 and PGC-1α. In conclusion, these results indicated that polyphenols stimulate browning in adipocytes via activation of the ß3-AR/AMPK signaling pathway, and SCLP and its monomer may be worth investigating to prevent obesity.

Eco-Stoichiometric Characteristics of Rhizosphere and Bulk Soils of Smilax china L. along Vertical Zone Spectrum of Fanjing Mountain.

To explore the correlations between nutrients and stoichiometric characteristics in the rhizosphere and bulk soils of understory Smilax china L. in forest ecosystems at different altitudes and to clarify the rhizosphere effect of understory vegetation in forest ecosystems and its response strategy to altitude, providing a theoretical basis for better forest ecological environment protection and high-quality development in Fanjing Mountain. Understory Smilax china L. at four different altitudes were selected, with the differences and influencing factors of carbon (C), nitrogen (N), phosphorus (P) and potassium (K) mass fractions and stoichiometric ratios in their rhizosphere and bulk soils analyzed. The average mass fractions of total C, total N and alkali-hydrolyzed N in the rhizosphere and bulk soils of Smilax china L. at different altitudes were 224.43 and 181.55 g·kg-1; 9.56 and 6.81 g·kg-1; and 648.19 and 600.70 g·kg-1, respectively. The rhizosphere effect of Smilax china L. was significant at altitudes of 500 m and 1000 m but became not so prominent with the rise of altitude. The C:N ratio in the rhizosphere and bulk soils ranged from 19.51 to 39.75 and the C:P ratio ranged from 225.29 to 543.05. C accumulation is greater than N accumulation in the rhizosphere and bulk soils of Smilax china L., and both present P limitation. Based on the comprehensive analysis of the mass fractions and eco-stoichiometric ratios of soil nutrients, the P limitation in Fanjing Mountain forest ecosystem is commonly seen and should be addressed.

The inhibitory kinetics and mechanism of quercetin-3-O-rhamnoside and chlorogenic acid derived from Smilax china L. EtOAc fraction on xanthine oxidase.

Smilax china L. showed various biological activities mainly due to its phenolic components; however, the mechanism of isolated phenolic fraction against xanthine oxidase (XO) has not been investigated. Quercetin-3-O-rhamnoside (QORh) and chlorogenic acid (CGA) extracted from Smilax china L. ethyl acetate fraction was analyzed for its XO inhibitory kinetics and mechanism using multispectroscopic methods and molecular docking techniques. QORh and CGA reversibly inhibited XO activity in competitive and non-competitive modes, respectively. The bioactive compounds bound with XO were dominated mainly by hydrogen bonds and van der Waals forces to form QORh-XO, and CGA-XO complexes with one affinity binding site. The synchronous fluorescence, circular dichroism, three-dimensional (3D) fluorescence, and Fourier transform infrared spectra exhibited that XO binding with QORh or CGA leads to the secondary and tertiary structural variation of the protein. Additionally, molecular docking further revealed that QORh binds to the active site of XO and forms hydrogen coupling with amino acid residues. The results showed that QORh and CGA had inhibitory activity on XO, which might be further used to modify the bioactive compounds and improve their efficacy to treat gout.

Smilax china Polyphenols Stimulate Browning via [Formula: see text]3-Adrenergic Receptor/AMP-Activated Protein Kinase [Formula: see text] Signaling Pathway in 3T3-L1 Adipocytes.

The aim of this study is to investigate the molecular mechanism of Smilax china L. polyphenols (SCLPs) in enhancing lipid metabolism and stimulating browning to reduce lipid accumulation in 3T3-L1 adipocytes. SCLP treatment obviously decreased lipid content in a dose-dependent manner (10-40 µg/mL) in adipocytes. SCLP treatment cooperated with noradrenalin to increase lipolysis. SCLPs reduced the gene expressions of C/EBP[Formula: see text] and Ap2 and enhanced the expressions of ACO, CPT, pHSL/HSL, ATGL, and PKA in adipocytes. Furthermore, SCLPs increased mRNA and protein expressions of brown adipocyte-specific factors (UCP-1, PRDM16, PGC-1α, and PPARγ) and mRNA expressions of beige adipocyte-specific markers (CD137, Tbx1, and Tmem26) in 3T3-L1 adipocytes, as well as mitochondrial biogenesis genes (Nrf1 and Tfam). In addition, according to the immunofluorescence staining, the mitochondria number was increased by SCLP. Moreover, ß3-AR or AMPK agonist synergistic SCLPs enhanced the expressions of UCP-1, PRDM16, and PGC-1α. While ß3-AR or AMPK antagonist significantly decreased the expressions of these brown adipocyte-specific factors, SCLP treatment inhibited the effect of antagonist to improve the expression of UCP-1, PRDM16, and PGC-1α. These results indicated that SCLPs may regulate lipid metabolism and stimulate browning via the ß3-AR/AMPKα signaling pathway. Thus, SCLPs likely have potential therapeutic effects on obesity.

Scopolin obtained from Smilax china L. against hepatocellular carcinoma by inhibiting glycolysis: A network pharmacology and experimental study.

ETHNOPHARMACOLOGICAL RELEVANCE: Smilax china L. is a well-known traditional medicinal plant. In China, it is a common anti-cancer drug that has been inherited for thousands of years. Some in vitro and in vivo studies have confirmed its potential lipid-lowering, anti-inflammatory and anti-ovarian cancer effects. However, there is no research on the material basis and mechanism of the rhizome of Smilax china L. against hepatocellular carcinoma. AIM OF THE STUDY: To explore the material basis and mechanism of scopolin from Smilax china L. against hepatocellular carcinoma. METHODS: The potential targets and active components of Smilax china L. against hepatocellular carcinoma were screened by transcriptomics, network pharmacology and molecular docking. Microscale Thermophoresis (MST) detection was used to verify the affinity of small molecule compounds with potential proteins and protein-protein interaction. The Extract from HepG2 cells was used to measure the expression of glycolysis-related proteins, glucose consumption and lactate production. The expression of apoptosis-related factors and glycolysis-related proteins in vivo was detected by immunohistochemistry. RESULTS: The glycolysis-related proteins glucose-6-phosphate isomerase (GPI), glycerol-3-phosphate dehydrogenase, mitochondrial (GPD2) and phosphoglycerate kinase 2 (PGK2) screened by transcriptomics, network pharmacology showed strongly binding with scopolin by molecular docking. MST detection has also verified the affinity of scopolin with GPI and GPD2. It was the first time found that Heat shock protein HSP 90-alpha (Hsp90α) bound strongly to GPI and GPD2 in the worldwide, while scopolin was able to affect the interaction between Hsp90α and GPD2. In vitro and in vivo experiments further demonstrated that scopolin may play an anti-cancer role by affecting the stability of tumor-associated proteins. The results showed that scopolin obtained from Smilax china L. could regulate the expression of GPI, GPD2 and PGK2 and inhibit the interaction of protein-protein, reduce the energy metabolism of tumor tissue, thereby inhibit tumor growth. CONCLUSION: Scopolin obtained from Smilax china L. plays the role of anti-hepatocellular carcinoma by regulating the expression of glycolysis proteins GPI, GPD2 and PGK2. Scopolin could affect the interaction between Hsp90α and GPD2 may provide a novel potential treatment direction for hepatocellular carcinoma.

Astilbin from Smilax glabra Roxb. alleviates high-fat diet-induced metabolic dysfunction.

Overweight, obesity, and related diseases are currently the major public health problems worldwide. Astilbin, extracted from the rhizome of Smilax glabra Roxb., is known to have significant anti-inflammatory activity and hepatoprotective effect. Studies have shown that it can inhibit adipogenesis in adipocytes in vitro; however, the intervention benefits of astilbin against obesity and related diseases along with its associated mechanisms remain unknown. This study aimed to demonstrate the impact of astilbin consumption on the overall biochemical pattern of high-fat diet (HFD) mice by using a combined multi-omics approach. Our data indicated that astilbin reduced body weight, insulin resistance, and inflammation in mice fed an HFD. Astilbin improved HFD-induced gut microbial dysbiosis by decreasing the Firmicutes-to-Bacteroidetes ratio, by increasing beneficial bacteria such as Alistipes and Muribaculum and decreasing harmful bacteria including Lachnospiraceae FCS020 group, Coriobacteriaceae UCG-002, and Lachnospiraceae UCG-008, resulting in enhanced intestinal carbohydrate and lipid metabolism. Meanwhile, astilbin protected the integrity of the intestinal barrier in HFD mice, increased short-chain fatty acid levels, and reduced metabolic endotoxemia. We further showed that astilbin attenuated hepatic lipid droplet aggregation and triglyceride accumulation in HFD mice, affected glutamate metabolism-related pathways, and enhanced hepatic ATP transduction pathways and attenuated xanthine metabolism pathways in mice, which were positively correlated with the abundance of Alistipes and negatively correlated with Ruminococcaceae UCG-003. The results highlighted that astilbin could be used as a prebiotic for the prevention of "gut-liver axis" damage and metabolic disruption in obese individuals.

New steroidal glycosides from the rhizomes of Smilax nipponica and their anti-inflammatory activities.

Herein, five new steroidal glycosides (Smilnipponicoside A-E) with five known congeners were isolated from the ethanol extracts of the rhizomes of Smilax nipponica Miq. Their structures were established by 2D NMR spectroscopic techniques (HMBC, HSQC, 1H,1H-COSY and NOESY), together with mass spectrometry, then comparison with the data in literature. The new compounds were subjected to evaluate anti-inflammatory in vitro. Compounds 1, 2 and 3 were found to have modest anti-inflammatory effects through suppression of TNF-α production in LPS-stimulated RAW 264.7 cells.

Protective effect of Smilax excelsa L. pretreatment via antioxidant, anti-inflammatory effects, and activation of Nrf-2/HO-1 pathway in testicular torsion model.

The protective effects of the ethanol extract of Smilax excelsa L. (SE) leaves were investigated on testicular tissue of rats with a torsion model in this study. The chemical composition of the extract was detected by means of liquid chromatography with tandem mass spectrometry (LC-MS/MS). SE extract was given for 21 days before torsion was created in the treatment group. The sperm parameters of the torsion group were impaired, and there was an increase in MDA level as well as a decrease in GSH level and GPx activity compared to the control group. TNF-α and NF-κB levels in the torsion group increased as compared to those in the control group. The expression levels of Nrf-2 and HO-1 were lower in the torsion group than those in the control group. The SE pretreatment group has improved sperm, oxidative stress, and inflammatory markers when compared to the torsion group, and the Nrf-2/HO-1 pathway was activated. PRACTICAL APPLICATIONS: Smilax excelsa L. is a plant with economic value used in traditional medicine in the treatment of stomachache, bloating, and breast cancer in Northwest Anatolia. It has an antioxidant effect due to the flavonoids and anthocyanins it contains. The protective effect against ischemia-reperfusion-induced tissue and reproductive damage in testicular tissue were demonstrated with the study. When the histological examinations of the tissues were evaluated, it was found that morphological structure of the tissues was retained in the treatment group. The findings indicate that SE prevents tissue damage in the torsion model by antioxidant and anti-inflammatory effects and activating Nrf-2/HO-1 pathway.

Smilax glabra Roxb. flavonoids protect against pathological cardiac hypertrophy by inhibiting the Raf/MEK/ERK pathway: In vivo and in vitro studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Smilax glabra Roxb., the dry rhizome of Sarsaparilla, which is also known as Tu fuling (TFL) in China, is a well-known traditional CHINESE medicine that is widely used for detoxication, relieving dampness and as a diuretic. We have previously shown that the extracted TFL flavonoids (designated TFLF) possess anti-cardiac hypertrophy effects in vitro. However, the anti-cardiac hypertrophy effects of TFLF in vivo and the underlying mechanisms remain to be elucidated. AIM OF THE STUDY: To reveal the underlying therapeutic mechanism of TFLF on cardiac hypertrophy by using transverse aortic constriction (TAC) model and cellular assays in vitro. MATERIAL & METHODS: Cardiac hypertrophy was replicated by TAC surgery in rats or by isoprenaline treatment of rat H9C2 myocardial cells in vitro. Cardiac structure and function were evaluated by echocardiographic and hemodynamic examinations in vivo and histological analysis of tissues ex vivo. Biochemical kits and quantitative PCR were used to analyze markers of cardiac hypertrophy. Expression and phosphorylation of key proteins in the Raf/MEK/ERK pathway were quantified by Western blotting. We further confirmed our findings in H9C2 rat cardiomyocytes treated with isoprenaline and the ERK inhibitor in vitro. RESULTS: TFLF attenuated cardiac hypertrophy and fibrosis and improved cardiac dysfunction in TAC rats. TFLF treatment induced a strong reduction in serum NT-proBNP levels. Cardiac hypertrophy marker gene (ANP, BNP and ß-MHC) expression and the phosphorylation levels of c-Raf and ERK1/2 were decreased by TFLF treatment. TFLF also protected H9C2 cells from isoprenaline-induced hypertrophy in vitro via a similar molecular mechanism as that observed in the rat heart. Moreover, pretreatment with TRLF and the ERK inhibitor further inhibited the mRNA overexpression of hypertrophic genes in vitro. CONCLUSIONS: TFLFs may protect against pathological cardiac hypertrophy via negative regulation of the Raf/MEK/ERK pathway. Thus, TFLFs are implicated as a potential pharmacological agent for treating cardiac hypertrophy in clinical practice.

Regulating the Imbalance of Gut Microbiota by Smilax china L. Polyphenols to Alleviate Dextran Sulfate Sodium-induced Inflammatory Bowel Diseases.

Smilax china L. is used not only as a kind of traditional Chinese herbal medicinal ingredients with various pharmacological properties, but also as food in certain parts of China. However, it is by far still unclear whether Smilax china L. polyphenols (SCP), as important bioactive constituents in Smilax china L., have effects on inflammatory bowel diseases (IBD). This study investigated the impact of SCP on the dextran sulfate sodium (DSS)-induced IBD and gut microbiota in mice. SCP treatments ameliorated typical symptoms of IBD as what was reflected through suppressing body weight loss, colonic shortening, intestinal barrier damage, and increasing intestinal disease activity index. SCP treatments simultaneously decreased the release of proinflammatory cytokines and oxidative stress, as well as promoted the release of anti-inflammatory factors. Furthermore, SCP ameliorated the ecological imbalance of gut microbiota and regulated the key bacteria associated with IBD (including Akkermansiaceae, Ruminococcaceae, Acidaminococcaceae, Muribaculaceae, and Anaeroplasmataceae). In general, SCP may improve DSS-induced IBD in mice by regulating inflammatory factors, inhibiting oxidative stress, reducing intestinal tissue damage, and regulating the ecological imbalance of intestinal microbiota. Thus, SCP might serve as a potential therapeutic agent against the inflammation-driven diseases.

Smilax china L. flavonoid alleviates HFHS-induced inflammation by regulating the gut-liver axis in mice.

BACKGROUND: Smilax china L., a traditional Chinese herb, has been used to treat various inflammatory disorders, particularly pelvic inflammation. The anti-inflammatory activity of the plant extract has been reported in several in vivo experimental models. However, the underlying anti-inflammatory mechanisms and the role of gut microbiota in mice on Smilax china L. flavonoid (SCF) treatment are poorly understand. PURPOSE: To investigate the role of SCF in providing the anti-inflammatory response and the role of gut microbiota in high-fat/high-sucrose (HFHS)-induced obese mice for 12 weeks. STUDY DESIGN AND METHODS: C57BL/6J mice were randomly divided into seven groups, normal chow (NC), HFHS, Orlistat, SCE, and low-, medium-, high- doses of SCF for 12 weeks. The body weight, liver weight, serum concentrations of lipopolysaccharide (LPS), and inflammatory cytokines in mice were assessed. The gene and protein expression levels of inflammation-related markers were measured by qRT-PCR and Western blot. Finally, the composition of gut microbiota was detected by analyzing 16S rDNA gene sequences. RESULTS: SCF supplement reduced body weight gain, adipose tissue and liver indexes, attenuated serum levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, LPS, and increased IL-10, and adiponectin. SCF significantly reduced the mRNA expression levels of TNF-α, IL-6, and increased the expression of AMPK, PPAR-γ, and IL-10 in mice's liver and adipose tissues. In addition, the TLR4, p-IκBα, NF-κB, and p65 protein expression levels were reduced after the SCF supplement. Moreover, SCF treatment ameliorated HFHS-induced gut dysbiosis, as revealed by an increased intestinal barrier protective species (Akkermansia spp). The relative abundance of Streptococcaceae, Faecalibaculum, and endotoxin-producing Desulfovibrionaceae were significantly decreased on SCF supplements. CONCLUSION: The results showed that SCF effectively inhibits HFHS-induced inflammation by suppressing the LPS-producing bacteria and pro-inflammatory bacteria group. Furthermore, the abundance of gut barrier protective species Akkermansia spp was increased to alleviate inflammatory response, inhibiting the LPS-TLR4/NF-κB signaling pathway. Thus, SCF may be a promising prophylactic for diet-induced inflammatory diseases through the gut-liver axis in mice.

Pectic polysaccharide from Smilax china L. ameliorated ulcerative colitis by inhibiting the galectin-3/NLRP3 inflammasome pathway.

Ulcerative colitis (UC) is an inflammatory bowel disease that affects the colon and rectum. Although galectin-3 (Gal-3) has been reported to play a proinflammatory role in UC, it is unknown whether pectic polysaccharide, a Gal-3 inhibitor in tumor metastasis, can alleviate UC by inhibiting Gal-3. The aim of this study was to investigate the anti-inflammatory effects and underlying mechanisms of SCLP, a pectic polysaccharide purified from Smilax china L. in our previous work, on dextran sulfate sodium-induced UC in BALB/c mice. The results showed that SCLP could significantly improve symptoms, alleviate histopathological damage and reduce the secretion of inflammatory mediators in mice with UC. Analysis of the anti-colitis mechanisms indicated that SCLP could inhibit the Gal-3/NLRP3 inflammasome/IL-1ß pathway by suppressing the expression of Gal-3 and the interaction of Gal-3 and NLRP3. Our results suggested that SCLP could be a promising candidate for prevention and treatment of UC.