Prevention of UVB-Induced Photoaging by an Ethyl Acetate Fraction from Allomyrina dichotoma Larvae and Its Potential Mechanisms in Human Dermal Fibroblasts.

Allomyrina dichotoma larvae (ADL) is an insect type that is used ethnopharmacologically to treat various diseases; however, its use as an antiaging treatment has not been widely studied. Previously, we found that an ethyl acetate (EA) fraction derived from an ADL extract (ADLE) has a high polyphenol content and antioxidant properties. In this study, we identified the underlying molecular mechanism for the protective effect of the EA fraction against UVB-induced photodamage in vitro and ex vivo. UVB treatment increased intracellular reactive oxygen species levels and DNA damage; the latter of which was significantly decreased following cotreatment with the EA fraction. Biological markers of aging, such as p16INK4a, p21WAF1, and senescence-associated ß-gal levels, were induced by UVB treatment but significantly suppressed following EA-fraction treatment. UVB-induced upregulation of matrix metalloproteinase (MMP)-1 and downregulation of COL1A1 were also reversed by EA-fraction treatment in both cells and a 3D skin model, which resulted in increased keratin and collagen deposition. Moreover, EA-fraction treatment inhibited the phosphorylation of MAPKs (p38, ERK, and JNK) and nuclear factor (NF-)-kB and decreased the levels of inflammatory cytokines in UVB-treated cells. The results indicate that an EA fraction from ADLE ameliorates UVB-induced degradation of COL1A1 by inhibiting MMP expression and inactivating the MAPK/NF-κB p65/AP-1 signaling pathway involved in this process.

Identification of α-Glucosidase-Inhibitors in Edgeworthia gardneri (Wall.) Meisn. Using UPLC-Q-TOF-MS/MS Analysis.

Edgeworthia gardneri (Wall.) Meisn., a member of the genus Edgeworthia in the family Thymelaeaceae, has long been applied as an edible and medicinal plant in China. E. gardneria has a hypoglycemic effect and is used to prepare daily drinks for the prevention and treatment of diabetes. However, the hypoglycemic substances involved remain unknown. The present study aimed to screen the α-glucosidase-inhibitors of E. gardneri and analyze its chemical profile using a ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) method. As a result, the ethyl acetate fraction (EAF) had significant α-glucosidase-inhibitory and antioxidant activities but did not show an α-amylase-inhibitory activity. A total of 67 compounds were identified in the EAF by UPLC-Q-TOF-MS/MS analysis; among them, 48 compounds were first discovered in the genus Edgeworthia. Additionally, five flavonoids, namely, isoorintin, secoisolaricirinol, tiliroside, chrysin, and kaempferol, had α-glucosidase-inhibitory activities. Rutin had a α-amylase-inhibitory activity. Daphnoretin, a kind of coumarin, has α-glucosidase and α-amylase-inhibitory activities. These findings enrich the chemical library of E. gardneria. EAF has a selective α-glucosidase-inhibitory activity, and flavonoids and coumarins may be the active components of EAF. E. gardneria has important value for developing multiple-target hypoglycemic drugs.

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.

[Anti-inflammatory material basis and mechanism of Artemisia stolonifera based on UPLC-Q-TOF-MS combined with network pharmacology and molecular docking].

This study aimed to explore the anti-inflammatory material basis and molecular mechanism of Artemisia stolonifera based on the analysis of the chemical components in different extracted fractions of A. stolonifera and their antioxidant and anti-inflammatory effects in combination with network pharmacology and molecular docking. Thirty-two chemical components were identified from A. stolonifera by ultra-performance liquid chromatography coupled to tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Among them, there were 7, 21 and 22 compounds in water, n-butanol and ethyl acetate fractions, respectively. The antio-xidant capacity of different extracted fractions was evaluated by measuring their scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) free radicals and total antioxidant capacity [ferric reducing antioxidant power(FRAP) assay]. The inflammatory model of RAW264.7 cells was induced by lipopolysaccharide(LPS), and the levels of nitrite oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) in the supernatant and the mRNA expression of related inflammatory factors in cells were used to evaluate the anti-inflammatory effects. The results revealed that ethyl acetate fraction of A. stolonifera was the optimal antioxidant and anti-inflammatory fraction. By network pharmacology, it was found that flavonoids such as rhamnazin, eupatilin, jaceosidin, luteolin and nepetin could act on key targets such as TNF, serine/threonine protein kinase 1(AKT1), tumor protein p53(TP53), caspase-3(CASP3) and epidermal growth factor receptor(EGFR), and regulate the phosphatidylinositol-3-kinase-protein kinase B(PI3K-AKT) and mitogen-activated protein kinase(MAPK) signaling pathways to exert the anti-inflammatory effects. Molecular docking further indicated excellent binding properties between the above core components and core targets. This study preliminarily clarified the anti-inflammatory material basis and mechanism of ethyl acetate fraction of A. stolonifera, providing a basis for the follow-up clinical application of A. stolonifera and drug development.

Microencapsulated antioxidant stevia fraction fortifies whey protein and enhances its antidiabetic activity.

Whey protein was fortified with a microencapsulated fraction of Stevia rebaudiana, in the proportion 1:4 (w/w), with maltodextrin from the elite variety of Stevia UEM-13, rich in antioxidant compounds, and evaluated its antioxidant and antidiabetic potential in vitro. The fraction in ethyl acetate, the microencapsulated fraction, the whey protein obtained by membrane and a commercial whey protein were characterized and were also investigated solubility, microencapsulation efficiency and stability and digestion in vitro. In addition, these products and two formulations of the icroencapsulated fraction with the obtained whey protein were tested for their potential to inhibit the α-amylase and α-glucosidase enzyme (antidiabetic activity). The microencapsulated fraction (0.5%) and the supplement fortified with the 20% fraction microencapsulated showed inhibitory potential for the enzyme. As for the α-glucosidase enzyme, all products tested showed inhibition, with the formulation with 1.6% microencapsulated fraction added to whey protein being significantly higher. The microencapsulated fraction showed better solubility and stability, including in vitro digestion analysis, and showed antioxidant and antidiabetic capacity. A sensory evaluation was performed with panelists who regularly consume whey protein supplements and products with stevia and the supplement formulation with 1.6 g microencapsulated stevia per 100 g of whey protein have good sensory acceptance.

Elsholtzia rugulosa: Phytochemical Profile and Antioxidant, Anti-Alzheimer's Disease, Antidiabetic, Antibacterial, Cytotoxic and Hepatoprotective Activities.

Elsholtzia rugulosa Hemsl., a species of the Labiatae family, has a long history of use as a honey plant, herbal tea, and folk medicine in China. However, little is known about its composition and biological activities. The present study aimed to investigate the total phenol and flavonoid contents, phytochemical composition, and multiple biological activities of this plant. The total flavonoid content of the ethyl acetate fraction (EAF) was higher than those of the petroleum ether fraction (PEF), n-butanol fraction (NBF), and water fraction (WF). The EAF also had much stronger antioxidant, cytotoxic, hepatoprotective, and acetylcholinesterase (AChE) and α-glucosidase inhibitory activities than the PEF, NBF, and WF. More importantly, the IC50 values of the EAF and NBF against α-glucosidase were much lower than that of the positive control acarbose, indicating their potent α-glucosidase inhibitory activities. The isolation of the EAF led to the acquisition of 9 compounds, four of which (ß-daucosterol, methyl rosmarinate, betulinic acid, and oleanolic acid) possessed significant α-glucosidase inhibitory activities. Maltol 6'-O-(5-O-p-coumaroyl)-ß-D-apiofuranosyl-ß-D-glucopyranoside and rosmarinic acid were the major phenolic compounds in the EAF according to the HPLC-DAD analysis. All these findings indicate that the EAF, NBF, and some isolated compounds have the potential to be developed as antidiabetic drugs. Moreover, the dual inhibition of AChE and butyrylcholinesterase (BChE) of certain fractions indicates their potential in the development of anti-Alzheimer's disease drugs. The present study provides a new understanding of the phytochemistry and bioactivity of E. rugulosa.

Effect of Aqueous and Ethyl Acetate Fractions of Ziziphus jujuba Mill Extract on Cardiovascular Responses in Hypertensive Rats.

BACKGROUND: Ziziphus jujuba Mill (ZJ) is a plant with anti-hypertensive property. In this regard, the present study investigated the effect of aqueous and ethyl acetate fractions of ZJ extract on acute hypertension (HTN) induced by nitro-L-arginine methyl ester (L-NAME). METHODS: The current study was carried on 49 hypertensive rats divided into seven groups, including i) control; ii) L-NAME (10 mg/kg); iii) sodium nitroprusside (SNP) (50 µg/kg) plus L-NAME; iv and v) aqueous fraction of ZJ (150 mg/kg and 300 mg/kg) plus L-NAME; vi) and vii) ethyl acetate fractions of ZJ (150 mg/kg and 300 mg/kg) plus L-NAME. The rats were orally treated with both fractions for four weeks and received intravenous L-NAME on the 28th day. The mean arterial pressure (MAP), systolic blood pressure (SBP) and heart rate (HR) of the rats were recorded then maximal changes (Δ) of MAP, SBP and HR were calculated and compared with changes of control and L-NAME. RESULTS: According to the obtained results of the present study, it was shown that the administration of L-NAME significantly increased ΔMAP, ΔSBP and ΔHR, and these effects were significantly attenuated by administration of SNP. The pre-treatment with both doses (150 mg/kg and 300 mg/kg) of aqueous and ethyl acetate fractions could significantly reduce cardiovascular responses induced by L-NAME that comparable with SNP. However, a lower dose of aqueous fractions and higher dose of ethyl acetate fractions were reported with stronger effects. CONCLUSION: The results of the current study showed that both the aqueous and ethyl acetate fractions of ZJ through the effect on nitric oxide system can prevent the development of HTN induced by L-NAME.

In Vitro Antioxidant and Anti-Propionibacterium acnes Activities of Cold Water, Hot Water, and Methanol Extracts, and Their Respective Ethyl Acetate Fractions, from Sanguisorba officinalis L. Roots.

Identification of medicinal plants and naturally derived compounds as new natural antioxidant and antibacterial sources for topical acne treatment has long been important. To determine anti-Propionibacterium acnes activity and in vitro antioxidant activities, Sanguisorba officinalis L. root (SOR) was extracted with cold water (CWE), hot water (HWE), and methanol (ME), and each extract was fractionated successively with hexane, ethyl acetate (EA), and butanol to determine whether the activities could be attributed to the total phenolic, flavonoid, terpenoid, and condensed tannin contents. Pearson's correlation coefficients were analyzed between the respective variables. The SOR CWE, HWE, ME, and their respective EA fractions showed anti-P. acnes activity based on the paper disc diffusion method on agar plates, minimum inhibitory concentration (MIC), and minimal bactericidal concentration (MBC). The MIC against P. acnes had a moderate (+) correlation with the total phenolic content, but not with the other measures. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging capacity (SC) had a strong (⁻) correlation with the total phenolic content and a moderate (⁻) correlation with the total flavonoid content. The total antioxidant capacity had a strong (+) correlation with the condensed tannin content. Linoleic acid peroxidation inhibition had a strong (⁻) correlation with the total phenolic content. To elucidate the major active phytochemicals in the CWE-EA, HWE-EA, and ME-EA fractions, high performance liquid chromatography-ultraviolet (HPLC-UV) and ultra high performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) were performed. The HPLC-UV analysis showed the presence of nine compounds in common (arjunic acid and/or euscaphic acid, gallic acid, kaempferol, caffeic acid, ferulic acid, tannic acid, and coumarin, quercetin). The UHPLC-QTOF-MS analysis showed the presence of nine compounds in common (gallic acid; caffeic acid; umbelliferone; arjunic acid, euscaphic acid, and/or tormentic acid; pomolic acid; rosamultic acid; and benzoic acid). When standards of the identified phytochemicals were tested against the same bacterium, quercetin, coumarin, and euscaphic acid showed antibacterial activity against P. acnes.

The gastroprotective effect of ethyl acetate fraction of hot water extract of Trichosanthes cucumerina Linn and its underlying mechanisms.

BACKGROUND: Antacids, anticholinergic drugs, histamine H2- receptor antagonists and irreversible proton pump inhibitors have been used for the treatment of gastric ulcers. However, prolonged use of these drugs may lead to series of adverse effects such as diarrhea, headache, rash, hypertension, muscular and joint pain. Therefore, there is an urgent need of more effective and safer treatments with fewer side effects. The aim of the present study was to scientifically evaluate the gastroprotective activity of fractions of the hot water extract of Trichosanthes cucumerina Linn (Family: Cucurbitaceae) aerial parts with a view to identifying the fraction with the best gastroprotective activity and the possible mechanism/s by which this fraction exert gastroprotection. METHODS: Gastroprotective activity of hexane fraction (HF), ethyl acetate fraction (EF), butanol fraction (BF) and aqueous fraction (AF) were evaluated by the assessment of ability to reduce the ulcer index in ethanol-induced rat model and the mode of action by which the most active fraction mediating gastroprotection. RESULTS: EF showed the maximum gastroprotection effect followed by BF and AF. EF (75 mg/kg) exhibited significantly higher gastroprotection compared to the reference drugs. Further investigations with two lower doses of EF confirmed that EF can mediated a significant and dose dependent gastroprotection. The rats treated with the EF showed significant reduction in free acidity (45%), total acidity (by 48%) in the gastric juice, increased the amount of mucus produced by the rat gastro mucosa and potent antihistamine activity (by 25.6%). EF was also rich in phenolic compounds and flavonoids. CONCLUSION: Gastroprotective mechanism of EF is possibly involves inhibition of acidity, elevation in mucus content, inhibition of histamine and antioxidant mechanisms.

Polyphenol-Rich Fraction from Larrea divaricata and its Main Flavonoid Quercetin-3-Methyl Ether Induce Apoptosis in Lymphoma Cells Through Nitrosative Stress.

Larrea divaricata is a plant with antiproliferative principles. We have previously identified the flavonoid quercetin-3-methyl ether (Q-3-ME) in an ethyl acetate fraction (EA). Both the extract and Q-3-ME were found to be effective against the EL-4 T lymphoma cell line. However, the mechanism underlying the inhibition of tumor cell proliferation remains to be elucidated. In this work, we analyzed the role of nitric oxide (NO) in the induction of apoptosis mediated by Q-3-ME and EA. Both treatments were able to induce apoptosis in a concentration-dependent and time-dependent manner. The western blot analysis revealed a sequential activation of caspases-9 and 3, followed by poly-(ADP-ribose)-polymerase cleavage. EA and Q-3-ME lowered the mitochondrial membrane potential, showing the activation of the intrinsic pathway of apoptosis. Q-3-ME and EA increased NO production and inducible NO synthase expression in tumor cells. The involvement of NO in cell death was confirmed by the nitric oxide synthases inhibitor L-NAME. In addition, EA and Q-3-ME induced a cell cycle arrest in G0/G1 phase. These drugs did not affect normal cell viability. This data suggested that EA and Q-3-ME induce an increase in NO production that would lead to the cell cycle arrest and the activation of the intrinsic pathway of apoptosis. Copyright © 2016 John Wiley & Sons, Ltd.

Bioprospecting the Curculigoside-Cinnamic Acid-Rich Fraction from Molineria latifolia Rhizome as a Potential Antioxidant Therapeutic Agent.

Increasing evidence from both experimental and clinical studies depicts the involvement of oxidative stress in the pathogenesis of various diseases. Specifically, disruption of homeostatic redox balance in accumulated body fat mass leads to obesity-associated metabolic syndrome. Strategies for the restoration of redox balance, potentially by exploring potent plant bioactives, have thus become the focus of therapeutic intervention. The present study aimed to bioprospect the potential use of the curculigoside-cinnamic acid-rich fraction from Molineria latifolia rhizome as an antioxidant therapeutic agent. The ethyl acetate fraction (EAF) isolated from M. latifolia rhizome methanolic extract (RME) contained the highest amount of phenolic compounds, particularly curculigoside and cinnamic acid. EAF demonstrated glycation inhibitory activities in both glucose- and fructose-mediated glycation models. In addition, in vitro chemical-based and cellular-based antioxidant assays showed that EAF exhibited high antioxidant activities and a protective effect against oxidative damage in 3T3-L1 preadipocytes. Although the efficacies of individual phenolics differed depending on the structure and concentration, a correlational study revealed strong correlations between total phenolic contents and antioxidant capacities. The results concluded that enriched phenolic contents in EAF (curculigoside-cinnamic acid-rich fraction) contributed to the overall better reactivity. Our data suggest that this bioactive-rich fraction warrants therapeutic potential against oxidative stress-related disorders.

The regulatory effects of Cissus quadrangularis on some enzymes involved in carbohydrate metabolism in streptozotocin-induced diabetic rats.

CONTEXT: Diabetes mellitus (DM) is probably the single most important metabolic disease and is widely recognized as one of the leading causes of death and disability. Cissus quadrangularis Linn. (Vitaceae) is a medicinal food and is reported to possess hypoglycemic activity. OBJECTIVE: The present study evaluates the effect of ethyl acetate fraction of C. quadrangularis stem (CQSF) on carbohydrate metabolism in hepatic tissues of experimental diabetic rats. The phytochemical compounds present in the CQSF extract were identified by gas chromatography-mass spectrometry (GC-MS) analysis. MATERIALS AND METHODS: Diabetic animals were treated with CQSF (50, 100, and 200 mg/kg body weight) for 45 d. Several indices such as blood glucose, glycated hemoglobin (HbA1c), insulin, liver function tests, hepatic glycogen content, and the activities of carbohydrate-metabolizing enzymes were assayed after 45 d of extract treatment. RESULTS: A pronounced effect was observed with extract doses 100 and 200 mg/kg body weight. CQSF at a dose of 100 mg/kg body weight significantly decreased the altered levels of blood glucose by about 56%. CQSF also modulated the activities of carbohydrate-metabolizing enzymes by significantly increasing the activity of hexokinase (1.9-fold) and pyruvate kinase (2.2-fold) and significantly reducing the activity of glucose-6-phosphatase (41.23%), fructose-1,6-diphosphatase (29.43%), and glycogen phosphorylase (35.07%). GC-MS analysis revealed the presence of 10 chemical constituents, and N-methyl-1-adamantane acetamide was found to be the prevailing compound in the extract. DISCUSSION AND CONCLUSION: The current study suggests the antidiabetic potential of CQSF, mediated through the regulation of carbohydrate metabolic enzyme activities.

Phytoconstituents of Nymphaea rubra flowers and their anti-diabetic metabolic targets.

Nymphaea rubra (N. rubra) flowers are prevalent in subtropical regions for both dietary and traditional medicinal purposes, attributing to their beneficial properties in supporting overall health. This study first time provides descriptions of the antidiabetic and dyslipidemic properties employing STZ induced high fat diet fed diabetic rats and inhibition of α-amylase enzyme activity first by in vitro analyses, followed by a confirmatory in silico study to create a stronger biochemical rationale. Furthermore, in 3 T3-L1 cells, this extract promoted the suppression of adipogenesis. GC-MS investigation of the ethyl acetate fraction of ethanolic extract of N. rubra flowers revealed the presence of marker compounds of N. rubra, Nuciferine, and Apomorphine, which were the focus of molecular docking studies. The acquired concentrations of Nuciferine (22.39%) and 10, 11-dimethoxy-Apomorphine (1.47%) were detected. Together with other alkaloids identified by GC-MS analysis from this extract, mechanistically suggested that it might be caused by the synergistic impact of these bioactive chemicals. Molecular docking has been done to check the binding affinities of various isolated phytochemicals with HPAA, the dose-response effect of 100 mg/kg and 250 mg/kg of flower extract after 30 days showed a significant effect on body weight, food, water intake, serum insulin, FBG, OGTT, lipid profile, glycated haemoglobin, liver and kidney function test. Kidney histopathology results show a significant effect. These findings offer a strong foundation for the potential application of the ethyl acetate fraction of ethanolic extract from Nymphaea rubra flowers and its bioactive constituent in an in vivo system for the treatment and control of diabetes and its associated condition dyslipidemia.

Drosophila melanogaster as a model organism for diabetes II treatment by the ethyl acetate fraction of Atriplex halimus L.

Type 2 diabetes (T2D) is the most common metabolic disorder. The undesirable effects of synthetic drugs demand a search for safe antidiabetic agents. This study aimed to assess the antidiabetic activity of different fractions of Atriplex halimus (petroleum ether 60-80, methylene chloride, ethyl acetate, and n-butanol) using Drosophila melanogaster larvae. Titers of total glucose and trehalose, as well as larval weight, were measured and compared with those of control and diabetic larvae. The expression of Drosophila insulin-like peptides (DILP2 and DILP3) and adipokinetic hormone (AKH) was evaluated. The results revealed a significant increase in total glucose, trehalose, and a decrease in body weight in the larvae fed a high-sugar diet compared with those in the control. When larvae fed diets containing the tested fractions, the total glucose and trehalose decreased to the control level, and the body weight increased. DILP2, DILP3, and AKH exhibited significant decreases upon treatment with A. halimus ethyl acetate. Metabolomic profiling of the ethyl acetate fraction of A. halimus revealed the presence of flavonoids and flavonoid glycosides. After docking screening to predict the most powerful moiety, we discovered that flavonoid glycosides (especially eriodictyol-7-O-neohesperidoside) have a greater affinity for the pocket than the other moieties. The results indicated the therapeutic activity of the A. halimus ethyl acetate fraction against induced T2D in Drosophila larvae. The antidiabetic activity may be attributed to flavonoids, which are the main components of the A. halimus ethyl acetate fraction.

In Vitro α-Amylase and α-Glucosidase Inhibitory Activity of Green Seaweed Halimeda tuna Extract from the Coast of Lhok Bubon, Aceh.

Seaweed belongs to marine biota and contains nutrients and secondary metabolites beneficial for health. This study aimed to determine the antidiabetic activity of extracts and fractions of green seaweed Halimeda tuna. The H. tuna sample was extracted with the maceration method using methanol and then partitioned using ethyl acetate and water to obtain ethyl acetate and water fractions. The methanol extract, ethyl acetate fraction, and water fraction of H. tuna were tested for their inhibitory activity against α-amilase and α-glucosidase. The methanol extract and the fractions with the highest inhibitory activity were phytochemically tested and analyzed using gas chromatography-mass spectrometry (GC-MS). The results showed that the ethyl acetate fraction (IC50 = 0.88 ± 0.20 mg/mL) inhibited α-amylase relatively similar to acarbose (IC50 = 0.76 ± 0.04 mg/mL). The methanol extract (IC50 = 0.05 ± 0.01 mg/mL) and the ethyl acetate fraction (IC50 = 0.01 ± 0.00 mg/mL) demonstrated stronger inhibitory activity against α-glucosidase than acarbose (IC50 = 0.27 ± 0.13 mg/mL). Phytochemical testing showed that the methanol extract and the ethyl acetate fraction contained secondary metabolites: alkaloids, flavonoids, steroids, and phenol hydroquinone. The compounds in methanol extract predicted to have inhibitory activity against α-amylase and α-glucosidase were Docosanol, Neophytadiene, Stigmasta-7,22-dien-3-ol,acetate,(3.beta.,5.alpha.,22E), Octadecanoic acid,2-oxo-,methyl ester, and phytol, while those in the ethyl acetate fraction were n-Nonadecane, Phytol, Butyl ester, 14-.Beta.-H-pregna, Octadecenoic acid, and Oleic acid.

The diuretic effect of ethyl acetate fractions of Artocarpus altilis, Artocarpus champeden, and Artocarpus heterophyllus leaves in normotensive Wistar rats.

BACKGROUND: Artocarpus altilis, A. Champeden, and Artocarpus heterophylus are popular species in Indonesia, and are commonly used as traditional medicine. OBJECTIVE: This study aims to evaluate the diuretic effects of the ethyl acetate fraction of these three species on normotensive Wistar rats. METHODS: The ethyl acetate fraction was prepared by a liquid-liquid extraction method. To evaluate diuretic effects, the sixty rats were divided into normal (distilled water), negative (4.5% NaCl), positive control (furosemide 5 mg/kg), and the testing groups. The testing groups were orally given the ethyl acetate fraction of A. altilis, Artocarpus champeden, and A. heterophylus at three dose levels of 25, 50, and 100 mg/kg. All animals were orally given 4.5% NaCl at a dose of 2 mL/200 g except the normal group; then the animals were given drugs according to group doses. Urine volume and electrolyte levels produced by the testing groups were compared to those of the control group. The concentration and ratio of ions were calculated to determine the natriuretic and carbonic anhydrase activities. RESULTS: The ethyl acetate fractions of the three Artocarpus species at 100 mg/kg dose were more active than the standard furosemide (p < 0.05) to increase urinary excretion. Furthermore, at doses of 50 and 100 mg/kg, the fraction significantly increased the excretion of Na+, K+, and Cl- ions more than the standard (p < 0.05). The testing groups showed good natriuretic and carbonic anhydrase activities. CONCLUSIONS: The ethyl acetate fraction of A. altilis, A. champeden, and A. heterophylus leaves performed good diuretic and natriuretic activities. Therefore, the fractions can be considered as potential natural diuretic medicines.

Anti-inflammatory, antinociceptive effects and involvement of opioid receptors in the antinociceptive activity of Eugenia uniflora leaves obtained with water, ethanol, and propylene glycol mixture.

ETHNOPHARMACOLOGICAL RELEVANCE: Eugenia uniflora (Myrtaceae) is a species native to Brazil and has a traditional use in the treatment of inflammation. AIM OF THE STUDY: To evaluate the anti-inflammatory and antinociceptive effects, and the involvement of opioid receptors in the antinociceptive activity of extract and fractions from Eugenia uniflora leaves. MATERIALS AND METHODS: TLC and HPLC were used to characterize the spray-dried extract (SDE) and fractions. In the in vivo assays, Swiss (Mus musculus) mice were used. Carrageenan-induced hind-paw edema and carrageenan-induced peritonitis models were used to determine the anti-inflammatory effect of the extract (50, 100, or 200 mg/kg). Acetic acid-induced writhing, tail-flick, and formalin tests were used to determine the antinociceptive effect of the extract (50, 100, or 200 mg/kg). The aqueous (AqF) and ethyl acetate (EAF) fractions (6.25, 12.5, and 25 mg/kg) were then combined with naloxone to evaluate the involvement of opioid receptors in the antinociceptive activity. RESULTS: In this work, the TLC and HPLC analysis evidenced the enrichment of EAF, which higher concentration of gallic acid (5.29 ± 0.0004 %w/w), and ellagic acid (1.28 ± 0.0002 %w/w) and mainly myricitrin (8.64 ± 0.0002 %w/w). The extract decreased the number of total leukocytes and neutrophils in the peritoneal cavity (p < 0.05), at doses of 100 and 200 mg/kg and showed significant inhibition in the increase of paw edema volume (p < 0.05). The treatment per oral route (doses of 50, 100, and 200 mg/kg) significantly reduced the nociceptive response in acetic acid-induced abdominal writhing (p < 0.05). The effect of the extract on the tail-flick test showed a significant increase in latency time of animals treated at doses of 200 and 100 mg/kg (p < 0.05). The extract and ethyl acetate fraction reduced the nociceptive effect in both phases of formalin at all tested doses. The naloxone reversed the antinociceptive effect of EAF, suggesting that opioid receptors are involved in mediating the antinociceptive activity of EAF of E. uniflora in the formalin test. CONCLUSION: The current study demonstrates the anti-inflammatory and analgesic activities of water: ethanol: propylene glycol spray-dried extract from E. uniflora leaves using in vivo pharmacological models in mice. Our findings suggest that spray-dried extract and ethyl acetate fraction exhibit peripheral and central antinociceptive activity with the involvement of opioid receptors that may be related to the presence of flavonoids, mainly myricitrin.

In Vitro Assessment of Organic and Residual Fractions of Nematicidal Culture Filtrates from Thirteen Tropical Trichoderma Strains and Metabolic Profiles of Most-Active.

The nematicidal properties of Trichoderma species have potential for developing safer biocontrol agents. In the present study, 13 native Trichoderma strains from T. citrinoviride, T. ghanense (2 strains), T. harzianum (4), T. koningiopsis, T. simmonsii, and T. virens (4) with nematicidal activity were selected and cultured in potato dextrose broth to obtain a culture filtrate (CF) for each. Each CF was partitioned with ethyl acetate to obtain organic (EA) and residual filtrate (RF) fractions, which were then tested on second-stage juveniles (J2s) of the nematodes Meloidogyne javanica and M. incognita in a microdilution assay. The most lethal strains were T. harzianum Th43-14, T. koningiopsis Th41-11, T. ghanense Th02-04, and T. virens Th32-09, which caused 51-100% mortality (%M) of J2s of both nematodes, mainly due to their RF fractions. Liquid chromatography-diode array detector-electrospray-high resolution mass spectrometry analysis of the most-active fractions revealed sesquiterpene and polyketide-like metabolites produced by the four active strains. These native Trichoderma strains have a high potential to develop safer natural products for the biocontrol of Meloidogyne species.

Xiaoyaosan ethyl acetate fraction alleviates depression-like behaviors in CUMS mice by promoting hippocampal neurogenesis via modulating the IGF-1Rß/PI3K/Akt signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Xiaoyaosan (XYS), a representative and classic traditional Chinese medicine (TCM) prescription with function of dispersing stagnated liver and strengthening spleen, has been used for thousands of years to treat depression. XYS' anti-depression effect has been demonstrated both clinically and experimentally; however, the material basis for this effect has yet to be elucidated. AIM OF THE STUDY: This study aimed to evaluate the impact and underlying action mechanism of XYS' antidepressant active component (Xiaoyaosan ethyl acetate fraction, XYSEF) against chronic unpredictable mild stress (CUMS)-induced depression-like behavior in mice. MATERIALS AND METHODS: First, we established a behavioral despair depression mouse model to preliminarily determine the effective antidepressant dose of XYSEF. Then, we created a CUMS mouse model and used various classic behavioral tests, including SPT, ST, NFST, and TST, to assess XYSEF's antidepressant properties. IGF-1 levels in mouse serum and hippocampus were quantified using ELISA. The average optical density of Nissl bodies in the mouse hippocampal CA3 region was determined utilizing toluidine blue staining. Brdu and DCX expression in the hippocampal dentate gyrus (DG) was assayed using the immunofluorescence method. IGF-1Rß, PI3K, p-PI3K, Akt, p-Akt, Caspase-3, and cleaved Caspase-3 protein levels in the hippocampus were determined with Western blot. RESULTS: The behavioral despair mouse model findings showed that 9.1 and 40 g/kg of XYSEF both significantly shortened the immobility time of mice, suggesting that the effective dose range was 9.1-40 g/kg. Compared to the CUMS mouse model, XYSEF at 20 and 40 g/kg markedly increased the sucrose preference percentage in the SPT and grooming time in the ST, shortened the immobility time in the TST and the feeding latency in the NSFT, and reversed the downregulated IGF-1 content in mouse serum and hippocampus. In addition, XYSEF amplified the average optical density of Nissl bodies in the hippocampal CA3 region, promoted Brdu and DCX expression in DG, and diminished IGF-1Rß, p-PI3K/PI3K, p-Akt/Akt, and cleaved Caspase-3/Caspase-3 protein levels in the hippocampi of CUMS mice. CONCLUSION: XYSEF acted as an antidepressant in mice exhibiting CUMS-induced depression-like behaviors, possibly by promoting hippocampal neurogenesis, reducing neuronal apoptosis, and inhibiting the over-activation of the IGF-1Rß/PI3K/Akt pathway.

Cytotoxicity and antiproliferative activity of ethanol and ethyl acetate fractions from polymeric nanoparticles of green tea leaves (Camellia sinensis) in breast cancer cell line MDA-MB-132.

Green tea (Camellia sinensis) has benefits. Its main potential content is epigallocatechin gallate, which has many bioactivity and pharmacological properties. However, herbal medicines have limitations on low solubility and stability. A nanoparticle delivery system is a perfect form of active ingredient development, because it can mediate the increase in solubility, dissolution rate, and strength of a targeted delivery system. This study aimed to make and test the formulation of the ethanol and ethyl acetate fraction from green tea leaves in the form of a nanoparticle delivery system using chitosan biopolymer as the primary carrier polymer combined with sodium tripolyphosphate as a crosslinker and then carried out the tests on the MDA-MB-231 breast cancer cell line. The results showed that the particle size value was 199.7 nm, the zeta potential was-56.7 mV, and the polydispersity index was 0.337. X-ray diffraction and differential scanning calorimetry test results showed that the C. sinensis fraction was perfectly dispersed molecularly in the nanoparticle system. The results of the cytotoxic test on the MDA-MB-231 breast cancer cell line obtained IC50 values for both fractions, namely 10.70 µg/mL (nano ethanol fraction) and 12.72 µg/mL (nano ethyl acetate fraction). This result showed a significant increase in anticancer activity in both fractions compared to those not formulated (P < 0.05). These results also show that the C. sinensis tea fraction formulated in a nanoparticle delivery system has a great potential as a new therapeutic agent for breast cancer.