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.

Probing interaction of a fluorescent ligand with HIV TAR RNA.

Trans-activator of Transcription (Tat) antagonists could block the interaction between Tat protein and its target, trans-activation responsive region (TAR) RNA, to inhibit Tat function and prevent human immunodeficiency virus type 1 (HIV-1) replication. For the first time, a small fluorescence ligand, ICR 191, was found to interact with TAR RNA at the Tat binding site and compete with Tat. It was also observed that the fluorescence of ICR 191 could be quenched when binding to TAR RNA and recovered when discharged via competition with Tat peptide or a well-known Tat inhibitor, neomycin B. The binding parameters of ICR 191 to TAR RNA were determined through theoretical calculations. Mass spectrometry, circular dichroism and molecular docking were used to further confirm the interaction of ICR 191 with TAR RNA. Inspired by these discoveries, a primary fluorescence model for the discovery of Tat antagonists was built using ICR 191 as a fluorescence indicator and the feasibility of this model was evaluated. This ligand-RNA interaction could provide a new strategy for research aimed at discovering Tat antagonists.

Two angiotensin-converting enzyme-inhibitory peptides from almond protein and the protective action on vascular endothelial function.

This study aimed to discover and prepare novel angiotensin converting enzyme (ACE) inhibitory peptides from almond protein and further evaluate the effect on endothelial function of human umbilical vascular endothelial cells (HUVECs). Almond protein was hydrolyzed using a two-stage alcalase-protamex hydrolysis process, and the hydrolysates were subjected to a series of separations, ultrafiltration, gel filtration chromatography, and reversed-phased preparative chromatography, to obtain the active peptides. Seven ACE inhibitory fractions with the molecular weight below 1.5 kDa were isolated and prepared, and two purified ACE inhibitory peptides with the IC50 values of 67.52 ± 0.05 and 43.18 ± 0.07 µg mL(-1), were identified as Met-His-Thr-Asp-Asp and Gln-His-Thr-Asp-Asp, respectively. Then the effect of two ACE inhibitory peptides on the endothelial function of HUVECs was evaluated. Results showed that the two potent ACE inhibitory peptides significantly regulated the release of nitric oxide and endothelin in HUVECs. These results suggest that almond peptides have potential as an antihypertensive nutraceuticals or a functional food ingredient.

Differential gene expression pattern in hypothalamus of chickens during fasting-induced metabolic reprogramming: functions of glucose and lipid metabolism in the feed intake of chickens.

Fasting-induced hypothalamic metabolic reprogramming is involved in regulating energy homeostasis and appetite in mammals, but this phenomenon remains unclear in poultry. In this study, the expression patterns of a panel of genes related to neuropeptides, glucose, and lipid metabolism enzymes in the hypothalamus of chickens during fasting and refeeding were characterized by microarray analysis and quantitative PCR. Results showed that 48 h of fasting upregulated (P < 0.05) the mRNA expressions of orexigenic neuropeptide Y and agouti-related protein but downregulated (P < 0.05) that of anorexigenic neuropeptide pro-opiomelanocortin; growth hormone-releasing hormone; islet amyloid polypeptide; thyroid-stimulating hormone, ß; and glycoprotein hormones, α polypeptide. After 48 h of fasting, the mRNA expression of fatty acid ß-oxidation [peroxisome proliferator-activated receptor α (PPARα), carnitine palmitoyltransferase 1A, and forkhead box O1], energy sensor protein [sirtuin 1 (SIRT1) and forkhead box O1], and glycolysis inhibitor (pyruvate dehydrogenase kinase, isozyme 4) were enhanced, but that of fatty acid synthesis and transport associated genes (acetyl-CoA carboxylase α, fatty acid synthase, apolipoprotein A-I, endothelial lipase, and fatty acid binding protein 7) were suppressed. Liver and muscle also demonstrated similar expression patterns of genes related to glucose and lipid metabolism with hypothalamus, except for that of acetyl-CoA carboxylase α, acyl-CoA synthetase long-chain family member 4, and apolipoprotein A-I. The results of intracerebroventricular (ICV) injection experiments confirmed that α-lipoic acid (ALA, pyruvate dehydrogenase kinase, isozyme 4 inhibitor, 0.10 µmol) and NADH (SIRT1 inhibitor, 0.80 µmol) significantly suppressed the appetite of chickens, whereas 2-deoxy-d-glucose (glycolytic inhibitor, 0.12 to 1.20 µmol) and NAD(+) (SIRT1 activator, 0.08 to 0.80 µmol) increased feed intake in chickens. The orexigenic effect of NAD(+) was also blocked by cotreatment with NADH. However, ICV injection of either GW7647 (PPARα agonist) or GW6471 (PPARα antagonist) showed no effects on feed intake. Results suggested that hypothalamic glycolysis (inhibited by ALA and promoted by 2-deoxy-d-glucose) and SIRT1 (inhibited by NADH and promoted by NAD(+)), not PPARα, were probably involved in feed intake regulation in chickens.

Aqueous biphasic system based on low-molecular-weight polyethylene glycol for one-step separation of crude polysaccharides from Pericarpium granati using high-speed countercurrent chromatography.

The aqueous biphasic system (ABS) plays a key role in the separation of bioactive substances, and the establishment and application of a low-molecular-weight polyethylene glycol (PEG) ABS remains a challenge in high-speed countercurrent chromatography (HSCCC). In this work, an ABS of low-molecular-weight PEG, namely PEG400-Na2SO4-H2O (20%-16%-64%, w/w/w), was developed on the basis of the phase diagram, and the phase forming time and ratio, and applied to HSCCC for the separation of polysaccharides. The crude polysaccharide extracted from Pericarpium granati (PGP) was successfully separated and three purified polysaccharides were obtained: PGP-1, with an average molecular weight of 13,210Da and composed of xylose (12.4%), ribose (10.1%), and glucose (77.5%); PGP-2, which is a homogeneous polysaccharide with an average molecular weight of 2584Da and consists of mannose; and PGP-3, with an average molecular weight of 2459Da and composed of ribose (51.4%), mannose (26.7%), and glucose (21.9%). This success shows that an ABS based on low-molecular-weight PEG could be applied to HSCCC separation technology.

Ultrasound-assisted extraction and preliminary purification of proanthocyanidins and chlorogenic acid from almond (Prunus dulcis) skin.

An aqueous solution of polyethylene glycol (PEG) as a green solvent was employed for the first time to develop the ultrasound-assisted extraction of proanthocyanidins (PA) and chlorogenic acid (CA) from almond skin. The optimized extraction parameters were determined based on response surface methodology, and corresponded to an ultrasound power of 120 W, a liquid-to-solid ratio of 20:1 (mL/g), and a PEG concentration of 50% (v/v). Under these optimized conditions, the extraction yields of PAs and CA from almond skin were 32.68 ± 0.22 and 16.01 ± 0.19 mg/g, respectively. Compared with organic solvent extraction, PEG solution extraction produced higher yields. Different macroporous resins were compared for their performance in purifying PAs and CA from almond skin extract. Static adsorption/desorption experimental results demonstrated that AB-8 resin exhibits excellent purification performance at pH 4. Under the optimized dynamic adsorption/desorption conditions on the AB-8 column, the total recovery of purification for PAs and CA was 80.67%. The total content of PAs and CA in the preliminarily purified extract was 89.17% (with respective contents of 60.90 and 28.27%).

Rapid analysis of fatty acid profiles in raw nuts and seeds by microwave-ultrasonic synergistic in situ extraction-derivatisation and gas chromatography-mass spectrometry.

Based on microwave-ultrasonic synergistic in situ extraction-derivatisation (MUED), gas chromatography-mass spectrometry was proposed for rapid analysis of fatty acid profiles in raw nut and seed materials. Several critical experimental parameters for MUED, including reaction temperature, microwave power, amounts of catalyst and derivatisation reagent, have been optimised using response surface methodology. The results showed that the chromatographic peak areas of total fatty acids and the content of total unsaturated fatty acids obtained with MUED were markedly higher than those obtained by the conventional method (P<0.05 and P<0.01, respectively). The MUED method simplified the handling steps compared to the conventional procedure, shortened the sample preparation time whilst improving the extraction and derivatisation efficiency of lipids, and reduced oxidisation and decomposition of the unsaturated fatty acids. The simplicity, robustness and practicality of this method highlighted its significant potential for application in the rapid analysis of fatty acids in natural food resource samples.

Extraction, antioxidant capacity and identification of Semen Astragali Complanati (Astragalus complanatus R. Br.) phenolics.

A study about the ultrasonic extraction, antioxidant capacity and identification of phenolics from Semen Astragali Complanati was undertaken. The optimised extraction condition with an orthogonal experiment design of L9(3(4)) was 50°C, 30min and 15:1 (methanol solution:sample, mL:g). An extraction yield of 40.12±1.10mg gallic acid/g dry seed and the antioxidant capacity of 0.85±0.13mg dry extract/mg DPPH were obtained with the optimum condition, respectively. For each extraction set, the antioxidant capacity by 2,2-diphenyl-1-picrylhydrazyl radical assay highly correlated to its total phenolic content by Folin-Ciocalteu method. Twelve phenolic components obtained with the optimised extraction procedure were tentatively identified by Ultra Performance Liquid Chromatography coupled with Q-TOF Mass Spectrometer (UPLC-Q-TOF) according to their mass spectrometry, UV/vis spectrometry, and related literature reports. Furthermore, the ultrasonic effect on the particles' microstructure of Semen Astragali Complanati was also investigated by scanning electron microscopy (SEM) analysis.

The anorexigenic effect of serotonin is mediated by the generation of NADPH oxidase-dependent ROS.

Serotonin (5-HT) is a central inhibitor of food intake in mammals. Thus far, the intracellular mechanisms for the effect of serotonin on appetite regulation remain unclear. It has been recently demonstrated that reactive oxygen species (ROS) in the hypothalamus are a crucial integrative target for the regulation of food intake. To investigate the role of ROS in the serotonin-induced anorexigenic effects, conscious mice were treated with 5-HT alone or combination with Trolox (a ROS scavenger) or Apocynin (an NADPH oxidase inhibitor) by acute intracerebroventricular injection. Both Trolox and Apocynin reversed the anorexigenic action of 5-HT and the 5-HT-induced hypothalamic ROS elevation. The mRNA and protein expression levels of pro-opiomelanocortin (POMC) were dramatically increased after ICV injection with 5-HT. The anorexigenic action of 5-HT was accompanied by markedly elevated hypothalamic MDA levels and GSH-Px activity, while the SOD activity was decreased. Moreover, 5-HT significantly increased the mRNA expression of UCP-2 but reduced the levels of UCP-3. Both Trolox and Apocynin could block the 5-HT-induced changes in UCP-2 and UCP-3 gene expression. Our study demonstrates for the first time that the anorexigenic effect of 5-HT is mediated by the generation of ROS in the hypothalamus through an NADPH oxidase-dependent pathway.

Microwave-assisted one-step extraction-derivatization for rapid analysis of fatty acids profile in herbal medicine by gas chromatography-mass spectrometry.

A rapid and practical microwave-assisted one-step extraction-derivatization (MAED) method was developed for gas chromatography-mass spectrometry analysis of fatty acids profile in herbal medicine. Several critical experimental parameters for MAED, including reaction temperature, microwave power and the amount of derivatization reagent (methanol), were optimized with response surface methodology. The results showed that the chromatographic peak areas of total fatty acids and total unsaturated fatty acids content obtained with MAED were markedly higher than those obtained by the conventional Soxhlet or microwave extraction and then derivatization method. The investigation of kinetics and thermodynamics of the derivatization reaction revealed that microwave assistance could reduce activation energy and increase the Arrhenius pre-exponential factor. The MAED method simplified the sample preparation procedure, shortened the reaction time, but improved the extraction and derivatization efficiency of lipids and reduced ingredient losses, especially for the oxidization and isomerization of unsaturated fatty acids. The simplicity, speed and practicality of this method indicates great potential for high throughput analysis of fatty acids in natural medicinal samples.

Three-dimensional cell bioreactor coupled with high performance liquid chromatography-mass spectrometry for the affinity screening of bioactive components from herb medicine.

An efficient and convenient method, three-dimensional (3-D) cell bioreactor coupled with high performance liquid chromatography-mass spectrometry was developed for affinity screening and analysis of multiple bioactive components from herbal medicines. Cancer cells were cultured on a porous scaffold to form a 3-D cell bioreactor. After interacting with live and fixed cells, the HPLC fingerprinting chromatograms of herbal medicine extract were compared to evaluate the binding properties of herbal components on cells. Model anticancer drugs (paclitaxel and resveratrol) and non-anticancer drugs (ketoprofen and penicillin G) were chosen to investigate the feasibility. When cell-drug interaction time was 30 min, the binding degrees of paclitaxel and resveratrol (each 15 µg/ml) were 82.2±7.2% and 66.1±4.1%, and for ketoprofen and penicillin G (each 15 µg/ml) were less than 3%. This method was used to screen bioactive components from Polygonum cillinerve (Nakai) Ohwi (PCO) extract, and the binding degrees of two main components in PCO extract (10 µg/ml), aristolochic acid A and aristolochic acid B, were 63.0±5.1% and 18.8±0.9%, respectively. These results demonstrated that this method was highly specific, efficient and convenient for affinity screening and analysis of bioactive components interacted with cells.

Protective effects of apricot kernel oil on myocardium against ischemia-reperfusion injury in rats.

The present study was undertaken to evaluate the potential cardioprotective effects of apricot kernel oil (AO) on the myocardial ischemia-reperfusion (IR) of rat model in vivo. The rats were divided into five groups: sham-operated, IR, low dose AO-treated IR (LD-AO+IR), medium dose AO-treated IR (MD-AO+IR) and high dose AO-treated IR (HD-AO+IR). All rats were provided with food and water ad libitum. The LD-AO+IR, MD-AO+IR and HD-AO+IR groups were given a daily dose of 2, 6 and 10 ml kg(-1)BW(-1) of AO, respectively, for 14 days prior to the IR operation. Tetrazolium chloride staining revealed that infarct size and the ratio of infarct weight to the total heart weight were decreased significantly in the three AO-treated groups compared to the IR group. The serum creatine kinase and aspartate aminotransferase activities also demonstrated similar beneficial effects. Myocardial catalase, superoxide dismutase, glutathione peroxidase, and constitutive nitric oxide synthase activities, as well as NO concentrations, were all increased, whereas malondialdehyde content and inducible nitric oxide synthase were decreased in AO-treated rats. These findings suggest that apricot kernel oil has potent cardioprotective effects, and could be developed as a nutriment for the treatment and prevention of myocardial infarcts.

Analysis of the interactions of multicomponents in Cornus officinalis Sieb. et Zucc. with human serum albumin using on-line dialysis coupled with HPLC.

Interactions of three iridoid glycosides extracted from Cornus officinalis Sieb. et Zucc. (CIG) with protein were simultaneously explored by on-line dialysis sampling coupled with high-performance liquid chromatography (DS-HPLC). Three main compounds in CIG were unequivocally identified as loganin, sweroside and cornuside by comparing their t(R), MS data and UV spectra with those of reference compounds. Dialysis recoveries and quantitative characteristics of DS-HPLC for three iridoid glycosides were determined. Recoveries of dialysis sampling ranged from 73.9 to 91.7% with the RSD below 3.0%. Based on the determination of concentrations before and after interaction with human serum albumin (HSA), the binding parameters of loganin, sweroside and cornuside with HSA were obtained and the binding mechanisms were investigated.

Analysis of fatty acids in A. szechenyianum Gay. by microwave-assisted extraction and gas chromatography-mass spectrometry.

INTRODUCTION: Aconitum szechenyianum Gay. is a traditional Chinese medicinal herb with the detumescent and styptic effects and antitumor activity. There have been only a few researches on its chemical components, but no detailed report has appeared on its fatty acids. OBJECTIVE: To develop a simple and effective method for the extraction of fatty acids from A. zechenyianum Gay. and then to investigate the fatty acid components. METHODOLOGY: Microwave-assisted extraction (MAE) was optimized with response surface methodology, and the fatty acid compositions of extract were determined by GC-MS with previous derivatisation to fatty acid methyl esters (FAMEs). The results were compared with that obtained by classical Soxhlet extraction (SE). RESULTS: Compared with SE, MAE showed significantly higher fatty acid yields, shorter extraction time, and lower energy and solvent consumption. The major fatty acids in A. szechenyianum Gay. are linoleic acid, palmitic acid, linolenic acid, oleic acid and stearic acid, and the unsaturated fatty acids occupy 66.4% of the total fatty acids.

Dialysis sampling on-line coupled with high-performance liquid chromatography for simultaneous investigation of the interactions between multi-components in herbs and the albumin.

An on-line dialysis sampling method coupled with high-performance liquid chromatography was developed for simultaneous investigation of the interactions between multiple bioactive compounds contained in herbal medicines and proteins. The system was used to estimate the interactions of components in danshen (Salvia miltiorrhiza) injection with bovine serum albumin. The results showed that the binding actions of five water-soluble compounds in danshen injection could be simultaneously investigated. The binding parameters of caffeic acid, ferulic acid, danshensu, protocatechuic acid and protocatechuic aldehyde were obtained and the interaction mechanisms were explored. The association constant evaluated for caffeic acid agreed well with literature values. The proposed approach should be beneficial for examining the holistic combined action of herbal medicine with proteins and help facilitate the discovery process of drug candidates.

Simultaneous determination of three Aconitum alkaloids in six herbal medicines by high-performance liquid chromatography.

To simultaneously determine three components of aconitine, mesaconitine, and hypaconitine in six species of Aconitum genus, an extraction condition for the total alkaloids was specifically optimized and a simple analytical method of reversed-phased highperformance liquid chromatography (HPLC) was developed. The extraction rate of total alkaloids in A. szechenyianum Gay was 98.3% for repeated extracting three times with an acidic alcohol solution (alcohol: pH 3.0 HAc = 85:15, v/v). The chromatography was carried out on a Phenomenex Luna C(18) column by gradient elution with a mobile phase of 0.03 mol/mL ammonium bicarbonate (pH = 9.50) -acetonitrile at a flow rate of 1.0 mL/min. The method for all three alkaloids had good linear relationships (r > 0.999) in the concentration range of 1.0-200.0 µg/mL. The average recoveries were 96.6-103.1%, and the LOQ and LOD were in the range of 25-37 ng/mL and 9-12 ng/mL, respectively. The quantitative results indicated that contents of the three alkaloids varied significantly among crude aconite roots, so quality control of traditional Chinese medicines containing aconite roots should be taken into account.

Anti-oxidation activity of the crude polysaccharides isolated from Polygonum cillinerve (Nakai) Ohwi in immunosuppressed mice.

AIM OF THE STUDY: Polygonum cillinerve (Nakai) Ohwi is commonly used in China for over 2000 years. Previous research has shown that the crude polysaccharides extracted from Polygonum cillinerve (Nakai) Ohwi (PCCP) have the scavenging free radicals and anti-tumor activities in vitro. In present study, PCCP were further approached the perspective of their anti-oxidation in immunosuppressed mice. METHODS AND MATERIALS: ICR mice were treated firstly with cyclophosphamide (CY, 150 mg/kg), 1 day later, treated with different dosages of PCCP or saline solution once daily for 21 days. Twenty-four hours later for the last drug administration, the animals were weighed, and then killed by decapitation. The liver, spleen, and thymus indices were investigated, and the biochemical parameters were evaluated for various tissues (liver, heart, and kidney). RESULTS: The administration of PCCP with gavage was able to overcome the cyclophosphamide-induced immunosuppression, and significantly to raise the TOC, CAT, SOD, and GSH-Px level. It also raised the liver, spleen, and thymus indices, and decreased the MDA level in mice. CONCLUSIONS: PCCP possess the pronounced free radical-scavenging and antioxidant activities, and could play an important role in the prevention of oxidative damage in immunological system.

LC-MS analysis for the components captured by ECV304 cell from extract of Aconitum szechenyianum Gay.

A novel method of cell affinity screening (CAS), cell affinity capture coupled with LC-MS analysis, was developed for screening the bioactive compounds related to cardiovascular diseases from the natural product libraries. One of the major characteristics lies in its function in affinity-capturing and separating the bioactive components from the natural product libraries in vitro. Another characteristic is its use in analyzing and identifying the target compounds, by employing high-performance liquid chromatography and mass spectrometry. CAS was used for screening the bioactive components from the alkaloid extract derived from Aconitum szechenyianum Gay. Of the five components found to be bound to the oxidative-damaged endothelial cells, the two compounds identified, mesaconitine and aconitine, were recognized in the literature as being related to cardiovascular diseases.

[Study on the optimum extracting conditions of total alkaloids and aconitine in Aconitum szechenyianum Gay].

OBJECTIVE: To optimize the conditions for extractions of total alkaloids and aconitine from Aconitum szechenyianum Gay. METHODS: Using L16 (4(5)) orthogonal test and univariant methods, the exrtaction conditions of total alkaloids aconitine in Aconitum szechenyianum Gay. were optimized under the guidance of the content determination of total alkaloids with bromophemol blue colorimetry and aconitine with HPLC method, respectively. RESULTS: The obtained optimum condition of total alkaloids was that the material was refluxed in 6 times (m/v) acidic alcohol solution( alcohol: pH 3.0 HAc = 85:15, v/v) for 1 h with 3 times. The condition of aconitine was that the material was refluxed in 4 times (m/v) acidic alcohol solution (alcohol: pH 3. p HAc = 15:85, v/v) for 0.5 h with 3 times. Contents of total alkaloids and aconitine in Aconitum szechenyianum Gay. were 0.980% and 0.109%, respectively. CONCLUSION: Through the test and verify, the optimum extraction methods of total alkaloids and aconitine are rational.

Cardiac muscle sarcolemma chromatographic stationary phase and its potential application in drug screening.

A new bioactive packing material for liquid chromatography, sarcolemma chromatography stationary phase (SCSP), is presented. Its surface characteristics are investigated, and it is found that the acceptors embedded in sarcolemma remained bioactive for more than a week. The retention behavior of antagonists and activators related to cardiac muscle sarcolemma on the SCSP chromatographic column shows the screening function of the SCSP column, and the retention behavior of the active components in the aether extract from the Chinese herb Ligusticum chuanxiong Hort. on the SCSP column reveals, to a certain extent, the separation function of the SCSP column. These suggest that SCSP is a potentially useful material in drug screening.