Enrichment of Bifidobacterium longum subsp. infantis ATCC 15697 within the human gut microbiota using alginate-poly-l-lysine-alginate microencapsulation oral delivery system: an in vitro analysis using a computer-controlled dynamic human gastrointestinal model

Abstract

This study evaluates alginate-poly-l-lysine-alginate Bifidobacterium longum subsp. infantis ATCC 15697-loaded microcapsules to enrich the human gut microbiota. The cell survival of alginate-poly-l-lysine-alginate microencapsulated B. infantis ATCC 15697 in gastric acid, bile, and through human gastrointestinal transit was investigated, as well as the formulation’s effect on the gut microbiota. Results show that microencapsulation increases B. infantis ATCC 15697 cell survival at pH1.0 (33.54?±?2.80% versus <1.00?±?0.00%), pH1.5 (41.15?±?2.06% versus <1.00?±?0.00%), pH2.0 (60.88?±?1.73% versus 36.01?±?2.63%), pH3.0 (75.43?±?1.23% versus 46.30?±?1.43%), pH4.0 (71.40?±?2.02% versus 47.75?±?3.12%) and pH5.0 (73.88?±?3.79% versus 58.93?±?2.26%) (p?<?0.05). In addition, microencapsulation increases cell survival at 0.5% (76.85?±?0.80% versus 70.77?±?0.64%), 1.0% (59.99?±?0.97% versus 53.47?±?0.58%) and 2.0% (53.10?±?1.87% versus 44.59?±?1.52%) (p?<?0.05) (w/v) bile. Finally, daily administration of alginate-poly-l-lysine-alginate microencapsulated B. infantis ATCC 15697 in a human gastrointestinal model induces a significant enrichment of B. infantis within the ascending (184.51?±?17.30% versus 53.83?±?17.82%; p?<?0.05), transverse (174.79?±?25.32% versus 73.17?±?15.30%; p?<?0.05) and descending (94.90?±?25.22% versus 46.37?±?18.93%; p?>?0.05) colonic microbiota.     

Sulfated polysaccharide isolated from Ulva lactuca attenuates d-galactosamine induced DNA fragmentation and necrosis during liver damage in rats

Context: Ulva lactuca Linnaeus (Chlorophyceae), a commonly distributed seaweed, is rich in polysaccharide but has not been studied extensively.

Objective: The present study investigated the effects of crude fraction of Ulva lactuca polysaccharide (ULP) on d-galactosamine (d-Gal)-induced DNA damage, hepatic oxidative stress, and necrosis in rats.

Materials and methods: The rats were treated with ULP (100?mg/kg, orally) for 4 weeks before a single intraperitoneal injection of d-Gal (500?mg/kg). In addition to liver cell necrosis and DNA damage, antioxidant parameters, such as lipid peroxide (LPO), superoxide dismutase, and catalase, and histopathology of liver tissue were evaluated.

Results: ULP pre-treatment significantly attenuated a d-Gal-induced decrease in DNA and RNA levels (3.67?±?0.38) and (5.42?±?0.46), respectively. Comet tail length and acridine staining confirmed the number of cells undergoing necrosis were relatively lower in ULP treated rats (30?µm and 8–10% of counted cells) compared to rats treated with d-Gal (60?µm and 16% of counted cells). Biochemical (LPO, SOD and CAT) and histological evaluation (p?d-Gal-induced elevation of LPO and infiltration of inflammatory cells into liver tissue.

Discussion and conclusion: Although our previous studies have reported on the protective role of ULP against liver toxicity, our present findings show that ULP improved the hepatic antioxidant defense system against d-Gal-induced DNA damage and necrosis in rats.     

Hesperidin,a citrus flavonoid,protects against l-methionine-induced hyperhomocysteinemia by abrogation of oxidative stress,endothelial dysfunction and neurotoxicity in Wistar rats

Context: Hesperidin (HSP), a flavanoglycone found in citrus fruits, has antioxidant, anti-inflammatory and neuroprotective properties.

Objective: This study evaluates the protective effect of HSP on l-methionine-induced hyperhomocysteinemia (HHcy) in rats.

Materials and methods: Male Wistar rats were randomly divided into seven groups as DMSO, l-methionine, HSP (25, 50 and 100?mg/kg), HSP-per se (100?mg/kg) and donepezil (0.1?mg/kg). HHcy was induced by oral administration of l-methionine (1.7?g/kg) for 32 days. From the 14^th day of study HSP (25, 50 and 100?mg/kg) and donepezil was administered orally to l-methionine-treated rats. Cognitive impairment induced by HHcy was determined using the Morris water maze (MWM) and Y-maze on video tracking system (28^th–32^nd day). Different biomarkers of HHcy in serum and brain and vascular reactivity were evaluated and histopathology (thoracic aorta and brain) was done.

Results: HSP (100?mg/kg) treatment in l-methionine-treated rats exhibited significant (p?p?l-methionine on acetylcholine-induced endothelial-dependent relaxation and increased serum nitrite and vascular nitric oxide bioavailability along with the restoration of histological aberrations.

Conclusion: HSP exerts a protective effect on HHcy by abrogating oxidative stress, ED and neurotoxicity.     

Hepatoprotective effect of Commiphora myrrha against d-GalN/LPS-induced hepatic injury in a rat model through attenuation of pro inflammatory cytokines and related genes

Abstract

Context: Commiphora myrrha (Burseraceae), a shrub resembling a small tree, has been used for several centuries for the treatment of various diseases.

Objective: This study investigates the hepatoprotective activity of C. myrrha ethanol extract against d-galactosamine/lipopolysaccharide (d-GalN/LPS)-induced acute hepatic injury in an animal model.

Materials and methods: Rats were pretreated with ethanolic extract C. myrrha (250 and 500?mg/kg; p.o.) for 7 d prior to the induction of an acute phase response by d-GalN/LPS. Animals were sacrificed 24?h after d-GalN/LPS (800?mg/kg and 50?µg/kg i.p.) administration for the biochemical and histological analyses.

Results: The administration of d-GalN/LPS increased plasma aminotransferases (174.47?±?4.5761 and 260.96?±?1.9839?µkat/l) and total bilirubin levels (1.012?±?0.0288?mg/dl), which were attenuated by C. myrrha treatment. Hepatic lipid peroxidation activity and nitric oxide content also increased, while the antioxidant activity measured by GSH (0.76 nmol/g protein), SOD (81.91?U/mg protein), and CAT (15.78?U/mg protein) was reduced. Commiphora myrrha provided significant restoration of GSH (0.815 nmol/gm protein), SOD (140.57?U/mg protein), and CAT (27.02?U/mg protein) levels. Furthermore, the acute phase response elicited by d-GalN/LPS administration enhanced mRNA expressions of TNF-α, IL-6, IL-10, iNOS-2, and HO-1, which were ameliorated by C. myrrha treatment.

Discussion and conclusion: These findings indicate that C. myrrha considerably reduces the oxidative stress of d-GalN/LPS-induced hepatic injury via multiple pathways including adown regulation of inflammatory mediators and cytokines. Such a property might be sufficient to combat cellular damage caused by various conditions that resemble fulminant hepatitis and could be of a potential clinical application.     

Intracerebroventricular administration of the (1→6)-β-d-glucan (lasiodiplodan) in male rats prevents d-penicillamine-induced behavioral alterations and lipoperoxidation in the cortex

Context: Lasiodiplodan, an exocellular (1→6)-β-d-glucan of molecular weight >1.4?×?10⁶?Da produced by MMPI strain of Lasiodiplodia theobromae (Pat.) Griffon &; Maubl. (Brotyosphaeriaceae) is known to exhibit anti-proliferative activity on breast cancer cells (MCF-7), anticoagulant activity when sulfonylated, and reduction in transaminase activity when administered in rats.

Objective: The effect of intracerebroventricular (I.C.V) injection of lasiodiplodan on neurotoxicity and behavioural changes induced by d-penicillamine was investigated.

Materials and methods: Twenty-four male Wistar rats were initially separated in groups of six and treated with 0.15?μmol/μL of NaCl (Groups Ct and d-Pen) and 0.01?μg/μL of lasiodiplodan (Groups Las and Las?+?d-Pen). After 15?min, they received 6?μmol/μL of NaCl (Groups Ct and Las) and 2?μmol/μL of d-penicillamine (Groups d-Pen and Las?+?d-Pen). The animal behavior was observed in an open-field test for 60?min. Twenty-four h later, the animals were sacrificed and histopathological analysis and Thiobarbituric acid reactive substances (TBARS) production measurements were performed.

Results: Lasiodiplodan prevented neurotoxicity induced by d-penicillamine significantly reducing the production of TBARS (308%; p?Discussion and conclusion: The reduction of TBARS production and convulsive episodes suggests that the protector effect provided by lasiodiplodan passes thought an antioxidant path, possibly interfering in a cascade of neurochemical events, triggering cell death and convulsive episodes. These results demonstrated that lasiodiplodan can be effective in treating neurotoxicity, and reducing damage triggered by convulsions in neuropathies related to GABAergic system.     

Mechanism of ginsenoside Rg1 renal protection in a mouse model of d-galactose-induced subacute damage

Context Ginseng is a widely used herbal medicine in China but its mechanism of action remains unclear.

Objective The objectives of this work were to study the protective effect of ginsenoside Rg1 on subacute murine renal damage induced by d-galactose and its mechanism.

Materials and methods C57BL/6J mice were injected with 120?mg/kg/d (sc) d-galactose for 1 week, followed by a combined treatment of Rg1 20?mg/kg/d (ip) and 120?mg/kg/d d-galactose (sc) for 5 weeks. Mice were injected with the 0.9% saline 0.2?mL/d (sc) and 120?mg/kg/d d-galactose (sc) for 6 weeks in the control group and the d-galactose group, respectively. After 6 weeks, urea, creatinine, uric acid, cystatin (Cys-C), senescence-associated β-galactosidase (SA-β-gal) staining positive kidney cells, superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malondialdehyde (MDA), glycation end products (AGEs) and 8-hydroxy-2 deoxyguanosine (8-OH-dG) were measured.

Results Treatment with Rg1 ameliorated kidney function and aging state (urea from 17.19?±?1.09 to 15.77?±?1.22?mmol·L?^??¹, creatinine from 29.40?±?5.72 to 22.60?±?3.97?μmol·L?^??¹, uric acid from 86.80?±?5.97 to 72.80?±?10.61?μmol·L?^??¹, Cys-C from 0.23?±?0.03 to 0.18?±?0.05?mg·L?^??¹, ROD of SA-β-gal from 56.32?±?10.48 to 26.78?±?7.34, SOD from 150.22?±?19.07 to 190.56?±?15.83 U·(mg·prot)?^?1, MDA from 9.28?±?1.59 to 3.17?±?0.82?nmol·(mg·prot)?^?1, GSH-PX from 15.68?±?2.11 to 20.32?±?2.96 U·(mg·prot)?^?1 as well as regulated glomerulus morphology (glomerulus diameter from 775.77?±?18.41 to 695.04?±?14.61?μm, renal capsule width from 39.56?±?3.51 to 31.42?±?2.70?μm, glomerulus basement membrane from 206.03?±?16.22 to 157.27?±?15.70?nm, podocyte slit from 55.21?±?8.55 to 37.63?±?6.65?nm).

Conclusions Ginsenoside Rg1 can antagonise d-galactose subacute renal damage in mice and this may occur due to alleviating oxidative stress injury.     

Chlorogenic acid protects d-galactose-induced liver and kidney injury via antioxidation and anti-inflammation effects in mice

Context Oxidative stress and inflammation are implicated in the aging process and its related hepatic and renal function decline. Chlorogenic acid (CGA) is one of the most abundant polyphenol compounds in the human diet. Recently, CGA has shown in vivo and in vitro antioxidant properties.

Objective The current study investigates the effects of protective effects of chlorogenic acid (CGA) on d-galactose-induced liver and kidney injury.

Materials and methods Hepatic and renal injuries were induced in a mouse model by subcutaneously injection of d-galactose (d-gal; 100?mg/kg) once a day for 8 consecutive weeks and orally administered simultaneously with CGA included in the food (200?mg/kg of diet). The liver and renal functions were examined. Histological analyses of liver and kidney were done by haematoxylin and eosin staining. The oxidative stress markers and pro-inflammatory cytokines in the liver and the kidney were measured.

Results CGA significantly reduced the serum aminotransferase, serum creatinine (SCr) and blood urea nitrogen (BUN) levels in d-gal mice (p?<0.05). CGA also restored superoxide dismutase, catalase, and malondialdehyde levels and decreased glutathione content in the liver and kidney in d-gal mice (p?<0.05). Improvements in liver and kidney were also noted in histopathological studies. CGA reduced tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) protein levels in the liver and kidney in d-gal mice (p?<0.05).

Discussion and conclusion These findings suggest that CGA attenuates d-gal-induced chronic liver and kidney injury and that this protection may be due to its antioxidative and anti-inflammatory activities.     

In vivo anti-inflammatory effect of a sulfated polysaccharide isolated from the marine brown algae Lobophora variegata

Context:?Lobophora variegata J.V. Lamouroux (Dictyotaceae) is a brown marine alga widely encountered in the Brazilian sea coast that presents high content of fucans. Anti-inflammatory effects of fucans are reported mostly in models in vitro, but little is known about its effects in vivo.

Objective:?To investigate vascular and cellular effects of a sulfated polysaccharide from the brown marine algae L. variegata (SP-Lv) in acute inflammatory models.

Materials and methods:?SP-Lv was isolated by DEAE-cellulose and analyzed by agarose gel electrophoresis and evaluated for its inhibitory effect on paw edema, vascular permeability, leukocyte migration and peritoneal nitrite content induced by zymosan in Wistar rats. Anticoagulant activities and possible systemic toxicity were also evaluated.

Results:?SP-Lv inhibited the paw edema (120 min: 1.42?±?0.11 vs. 0.95?±?0.05?mL), plasma exudation (21.53?±?0.62 vs. 11.96?±?0.68 μg/g), nitrite content (4.42?±?0.33 vs. 2.86?±?0.003 μM) and leukocyte migration (5.15?±?1.21 vs. 1.99?±?0.16 cells/10³ mL) induced by zymosan. SP-Lv and l-NAME reduced the paw edema (60–120?min) elicited by l-arginine. However, at 180?min SP-Lv effect was more accentuated and sustained until 240?min, while that of l-NAME was abolished. Similarly to indomethacin, SP-Lv inhibited the entire edema time-course induced by phospholipase A₂, except for the time of 60?min.

Discussion and conclusion:?The anti-edematogenic effect of SP-Lv seems to occur via inhibition of nitric oxide synthase and cyclooxygenase activities. These results suggest a potential applicability of polysaccharides from alga origin in acute inflammatory conditions.     

Effect of black mulberry (Morus nigra) extract treatment on cognitive impairment and oxidative stress status of d-galactose-induced aging mice

Context: Morus nigra L. (Moraceae) has various uses in traditional medicine. However, the effect of M. nigra on cognitive impairment has not been investigated yet.

Objective: The objective of this study is to determine the phenolic acid content and DNA damage protection potential of M. nigra leaf extract and to investigate the extract effect on cognitive impairment and oxidative stress in aging mice.

Materials and methods: Phenolic acid content was determined by quantitative chromatographic analysis. DNA damage protection potential was evaluated on pBR322 plasmid DNA. Thirty-two Balb-C mice were randomly divided into four groups (control, d-galactose, d-galactose?+?M. nigra 50, and d-galactose?+?M. nigra 100). Mice were administered d-galactose (100?mg/kg, subcutaneous) and M. nigra (50 or 100?mg/kg, orally) daily for 8 weeks. Behavioral responses were evaluated with Morris water maze. Activities of antioxidant enzymes and levels of malondialdehyde (MDA) were assayed in serum, brain, and liver.

Results: In extract, vanillic (632.093?μg/g) and chlorogenic acids (555.0?μg/g) were determined. The extract between 0.02 and 0.05?mg/mL effectively protected all DNA bands against the hazardous effect of UV and H₂O₂. Morus nigra significantly improved learning dysfunctions (p <?0.01), increased memory retention (p?<?0.01), reduced MDA levels (p?<?0.05), and elevated SOD, GPx, and CAT activities (p?<?0.05) compared with the d-galactose group.

Discussion and conclusion: These results show that M. nigra has the potential in improving cognitive deficits in mice and that M. nigra may be useful to suppress aging, partially due to its scavenging activity of free radicals and high antioxidant capacity.     

Compounds from the pods of Astragalus armatus with antioxidant,anticholinesterase, antibacterial and phagocytic activities

Context: The phytochemical study and biological activities of Astragalus armatus Willd. subsp. numidicus (Fabaceae) pods, an endemic shrub of Maghreb, are reported.

Objective: This study isolates the secondary metabolites and determines the bioactivities of Astragalus armatus pods.

Materials and methods: The chloroform, ethyl acetate and n-butanol extracts of hydro-ethanolic extracts were studied. Antioxidant activity was investigated using DPPH and ABTS radical scavenging, CUPRAC and ferrous chelating assays at concentrations ranging from 3 to 200?μg/mL. Anticholinesterase activity was determined against acetylcholinesterase and butyrylcholinesterase enzymes at 50, 100 and 200?μg/mL. Antibacterial activity was performed according to minimum inhibitory concentration (MIC) method. Carbon clearance method in albino mice was used for the phagocytic activity at concentrations 50, 70 and 100?mg/kg body weight. Spectroscopic techniques were used to elucidate the compounds.

Results: Ethyl acetate extract afforded a flavonoid (1) while the n-butanol extract gave four flavonoids (2–5), a cyclitol (6) and a cycloartane-type saponin (7). The ethyl acetate extract exhibited highest antioxidant activity in DPPH (IC₅₀: 67.90?±?0.57?μg/mL), ABTS (IC₅₀: 11.30?±?0.09?μg/mL) and CUPRAC (A_0.50: 50.60?±?0.9?μg/mL) assays. The chloroform extract exhibited the best antibacterial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, each with 80?μg/mL MIC values. The n-butanol extract enhanced phagocytic activity.

Discussion and conclusion: Isorhamnetin (1), isorhamnetin-3-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-galactopyranoside (2), isorhamnetin-3-O-β-d-apiofuranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 6)]-β-d-galactopyranoside (3), kaempferol-3-O-(2,6-di-O-α-l-rhamnopyranosyl)-β-d-galactopyranoside (4), kaempferol-3-O-(2,6-di-O-α-l-rhamnopyranosyl)-β-d-glucopyranoside (5), pinitol (6) and cyclomacroside D (7) were isolated whereas 1, 2, 6 and 7 are reported for the first time from A. armatus.     

Prevention of arsenic-mediated reproductive toxicity in adult female rats by high protein diet

Abstract

Context: The detrimental effects of arsenic on female reproductive functions may involve overt oxidative stress. Casein and pea [Pisum sativum Linn. (Fabaceae)] proteins have antioxidant properties.

Objective: To investigate the role of casein- and pea-supplemented high-protein diet (HPD) in utero-ovarian protection from arsenic toxicity.

Materials and methods: Adult female Wistar rats were orally gavaged with vehicle (Gr-I) or arsenic at 3?ppm/rat/d (Gr-II and Gr-III) for 30 consecutive days, when they were maintained on either regular diet containing 18% protein (Gr-I and Gr-II), or HPD containing 27% protein in the form of casein (20%) and pea (7%) (Gr-III). Reproductive functions were evaluated using a battery of biochemical and histological techniques.

Results: As compared to Gr-I, the Gr-II rats suffered from loss of estrous cyclicity, reduction in weight (mg/100?g body weight) of ovary (Gr-I: 54.3?±?4.2 versus Gr-II: 35.8?±?1.6; p?<?0.001) and uterus (Gr-I: 161.7?±?24.6 versus Gr-II: 94.44?±?13.2; p?<?0.05), utero-ovarian degeneration, attenuated ovarian activities (unit/mg tissue/h) of Δ⁵, 3β-hydroxysteroid dehydrogenase (Gr-I: 3.41?±?0.12 versus Gr-II: 2.31?±?0.09; p?<?0.01) and 17β-hydroxysteroid dehydrogenase (Gr-I: 3.82?±?0.57 versus Gr-II: 1.24?±?0.19; p?<?0.001), and decreased serum estradiol level (pg/ml) (Gr-I: 61.5?±?2.06 versus 34.1?±?2.34; p?<?0.001). Ovarian DNA damage was preponderant with blatant generation of malondialdehyde (nM/mg tissue; Gr-I: 15.10?±?2.45 versus Gr-II: 29.51?±?3.44; p?<?0.01) and attenuated superoxide dismutase activity (unit/mg tissue) (Gr-I: 2.18?±?0.19 versus Gr-II: 1.33?±?0.18; p?<?0.05). The Gr-III rats were significantly protected from these ill effects of arsenic.

Discussion and conclusion: HPD, by way of antioxidant properties, may find prospective role in the protection of reproductive damage caused by arsenic.     

Treated effect of silymarin on vascular function of aged rats: Dependant on nitric oxide pathway

Context: Aging leads to endothelial dysfunction and vascular stiffness which are the main causes of many cardiovascular diseases. Previous reports have shown that the cell protective effect of silymarin (SM) is dependent on its antioxidant properties.

Objectives: We investigated the effect of SM on vascular functions of aged rats and the involvement of nitric oxide or cyclooxygenase (COX) activity in this effect.

Materials and methods: Isolated rat aortas were obtained from 22-month old rats. Each ring was incubated with SM (50?mg/L), SM/l-nitro-arginine methyl ester (100?μM, l-NAME) or SM/indomethacin (10?μM, INDO) in tissue bath. Three- to four-month-old rats were used as young controls. Endothelium-intact rings were precontracted with α-receptor agonist phenylephrine (0.001–30?µM) or voltage-dependent high potassium (40?mM), endothelium dependent/independent relaxant responses were obtained using acetylcholine (0.001–30?µM) and sodium nitroprusside (0.0001–3?µM), respectively.

Results: Aging increased phenylephrine sensitivity (6.45?±?0.08; 6.88?±?0.09) and decreased KCl contraction (882?±?118.4; 499?±?80.4). SM treatment decreased the E_max of both agents (548?±?109; 223?±?48.9). Aging deteriorated acetylcholine relaxation (93.9?±?2.09; 72.0?±?2.56) and SM improved the response (86.3?±?1.90). l-NAME prevented the SM effect whereas INDO was ineffective.

Discussion and Conclusion: Immediate SM treatment partially restored endothelial dysfunction and vascular tone in aging. The possible mechanism might not be mediated by prostacyclin or the COX pathway in acute administration; the nitric oxide pathway and calcium antagonistic features of SM relate to its action on the vessel.     

Flavonoids,cytotoxic, antioxidant and antibacterial activities of Evax pygmaea

Context: Phytochemical study and biological potential of Evax pygmaea (L.) Brot. (Asteraceae) are reported for the first time.

Objective: To identify the secondary metabolites of Evax pygmaea and to determine its antioxidant, antibacterial and cytotoxic activities.

Materials and methods: Dried aerial parts (1?kg) were macerated in 70% MeOH (5?L) during 72?h. The concentrated hydromethanolic extract was subjected to extractions with chloroform (3?×?300?mL), ethyl acetate (3?×?300?mL) and n-butanol (3?×?300?mL), successively. VLC of combined ethyl acetate (EAEP) and n-butanol (BEP) fractions was followed by column purifications. Antioxidant activity was investigated using DPPH, CUPRAC, and metal chelating, β-carotene/linoleic acid and ABTS assays. Agar method was used in the antibacterial study. Cytotoxic activity was determined by Brine shrimp lethality test in DMSO and ethanol, at varying concentrations (2, 1 and 0.2%) and (1, 0.2 and 0.1%) successively.

Results: Quercetin (1), isorhamnetin 3-O-β-d-xyloside (2), isorhamnetin 3-O-β-d-glucoside (3), quercetin 3-O-β-d-glucoside (4), quercetin 7-O-β-D-glucoside (5), patuletin 3-O-β-d-glucoside (6) were isolated from for the first time from Evax genus. The EAEP was the most active in ABTS (IC₅₀: <3.125?μg/mL) assay whereas the BEEP exhibited the highest activity in the β-carotene/linoleic acid assay (IC₅₀: <3.125?μg/mL). The EAEP and BEP exhibited good antibacterial activity (MIC: 40–80 µg/mL). The plant did not show any toxicity (LD₅₀>80 µg/mL).

Discussion and conclusions: Six flavonoids were isolated for the first time from Evax pygmaea which exhibited good antioxidant and antibacterial activities.     

Antidiabetic potential of triterpenoid saponin isolated from Primula denticulate

Context: Primula denticulate Sm. (Primulaceae), commonly known as drumstick primula, is traditionally used to treat diabetes and urinary disorders. In the present study, a new triterpenoid saponin was isolated. Triterpenoids generally show antidiabetic activity. Considering its traditional use and chemical nature of the molecule, the present study was designed to evaluate the antidiabetic activity.

Objective: Antidiabetic activity of triterpenoid saponin (TTS) isolated from P. denticulate.

Materials and methods: A new TTS was isolated from the leaf of P. denticulate by column chromatography on CHCl₃/MeOH (8.5:1.5) fraction. It was further characterized by using NMR, UV, and IR spectroscopic methods. Ethanol and aqueous extracts of the leaf were also prepared. Antidiabetic study for TTS, ethanol extract, and aqueous extract was carried out in streptozotocin (STZ)-induced diabetic rats at doses of 200, 1000, and 1000?mg/kg body weight, respectively. A toxicity study was also performed.

Results: Isolated new TTS molecule was characterized as 3-O[β-d-xylopyranosyl-(1?→?2)-β-d-glucopyranosyl-(1?→?4)-α-l-arabinopyranosyloxy]-16α-hydroxy-13β,28-epoxy-olean-30-al by NMR, UV, and IR spectroscopic methods. This new TTS was found to be effective in lowering blood-glucose level in the experimental rat model, thus establishing its antidiabetic property (168.8?±?4.58) when compared with disease control (258.8?±?0.60). Its LD₅₀ value was found at a dose of 2000?mg/kg. The level of insulin was restored by TTS and ethanol extract up to 31.49?µU/ml and 38.90?µU/ml, respectively, when compared with disease control (18.45?µU/ml).

Discussion and conclusion: In conclusion, 3-O[β-d-xylopyranosyl-(1?→?2)-β-d-glucopyranosyl-(1?→?4)-α-l-arabinopyranosyloxy]-16α-hydroxy-3β,28-epoxy-olean-30-al possesses potential glucose lowering properties, i.e., antidiabetic potential against STZ-induced diabetic rats.     

Dual targeting of TNF-α and free radical toxic stress as a promising strategy to manage experimental polycystic ovary

Context: It is now clear that oxidative stress (OS) and chronic low-grade inflammation are two main pathways involved in polycystic ovary syndrome (PCOS) pathogenesis. Therefore, simultaneous targeting of these pathways by means of carvedilol and Semelil (ANGIPARS?), as established medicines with dual anti-cytokine and anti-oxidant potential may be a therapeutic alternative approach to the current treatments.

Objective: The objective of this study is to study the protective effects of carvedilol and ANGIPARS? on inflammatory and oxidative response in hyperandrogenism-induced polycystic ovary (PCO).

Materials and methods: The murine model of PCO was induced by letrozole (1?mg/kg/d; orally) and effective doses of carvedilol (10?mg/kg/d; orally) and ANGIPARS? (2.1?mg/kg/d; orally) were administrated for 21?d in PCO and non-PCO healthy rats. Ovarian folliculogenesis, sex hormones concentrations, OS, inflammatory, and metabolic biomarkers were assessed in serum and ovaries.

Results: PCO rats exhibited ovarian cystogenesis which was preserved by the application of carvedilol and ANGIPARS?. In comparison with controls, decreased level of the total antioxidant power (TAP) and higher levels of reactive oxygen species (ROS) and lipid peroxidation (LPO) in serum and ovaries (2.41?±?0.67 versus 0.72?±?0.11; and 0.17?±?0.04 versus 0.05?±?0.01; 5.48?±?1.30 versus 10.56?±?0.77; and 7.06?±?1.94 versus 17.98?±?0.98; p?<?0.05, respectively) were detected in PCO rats. Moreover, the PCO rats exhibited hyperandrogenism due to a 3.7-fold increase in serum testosterone concentration (35.04?±?3.17 versus 131.09?±?13.24; p?<?0.05) along with a 2.98-fold decrease in serum progesterone (6.19?±?0.40 versus 18.50?±?1.03; p?<?0.05) and 5.2-fold decrease in serum estradiol (9.30?±?0.61 versus 48.3?±?2.10; p?<?0.05) when compared with those of the control group. However, similar to the control group, normal levels of OS markers and sex hormones were detected in ANGIPARS? and carvedilol co-treated PCO rats. Besides, when compared with controls, increased levels of TNF-α (770.75?±?42.06 versus 477.14?±?28.77; p?<?0.05) and insulin (1.27?±?0.10 versus 0.36?±?0.05; p?<?0.05) in PCO rats were significantly inhibited by carvedilol and ANGIPARS? co-treatment.

Discussion and conclusion: We evidenced the beneficial effects of carvedilol and ANGIPARS? in PCO, which underpin the new alternative approach in using these kinds of medicines in female reproductive disorders.     

In vitro inhibition of UGT1A3, UGT1A4 by ursolic acid and oleanolic acid and drug–drug interaction risk prediction

1.?Ursolic acid (UA) and oleanolic acid (OA) may have important activity relevant to health and disease prevention. Thus, we studied the activity of UA and OA on UDP-glucuronosyltransferases (UGTs) and used trifluoperazine as a probe substrate to test UGT1A4 activity. Recombinant UGT-catalyzed 4-methylumbelliferone (4-MU) glucuronidation was used as a probe reaction for other UGT isoforms.

2.?UA and OA inhibited UGT1A3 and UGT1A4 activity but did not inhibit other tested UGT isoforms.

3.?UA-mediated inhibition of UGT1A3 catalyzed 4-MU-β-d-glucuronidation was via competitive inhibition (IC₅₀ 0.391?±?0.013?μM; K_i 0.185?±?0.015?μM). UA also competitively inhibited UGT1A4-mediated trifluoperazine-N-glucuronidation (IC₅₀ 2.651?±?0.201?μM; K_i 1.334?±?0.146?μM).

4.?OA offered mixed inhibition of UGT1A3-mediated 4-MU-β-d-glucuronidation (IC₅₀ 0.336?±?0.013?μM; K_i 0.176?±?0.007?μM) and competitively inhibited UGT1A4-mediated trifluoperazine-N-glucuronidation (IC₅₀ 5.468?±?0.697?μM; K_i 6.298?±?0.891?μM).

5.?Co-administering OA or UA with drugs or products that are substrates of UGT1A3 or UGT1A4 may produce drug-mediated side effects.     

Structural characterization of polysaccharides from the roots of Urtica fissa

Three polysaccharides were isolated from the roots of Urtica fissa by extraction, ultrafiltration, anion-exchange, and gel-filtration chromatography. The structures were characterized using acetylation, methylation, and spectral methods (GCMS, NMR). All three polysaccharides are mainly composed of d-arabinofuranosyl, d-galactopyranosyl, d-glucopyranosyl residues with different structural characteristics. Polysaccharide A of MW 5.2 × 10³ contained a linear chain of 1-linked β-d-glucopyranosyl, 1,6-linked β-d-glucopyranosyl, 1,6-linked α-galactopyranosyl, and 1,5-linked β-arabinofuranosyl moieties. Polysaccharide B of MW 7.7 × 10⁴ possessed a chain consisting of 1,5-linked α-d-arabinofuranosyl, 1,3-linked β-d-mannopyranosyl, 1,6-linked β-d-glucopyranosyl, and 1,6-linked α-d-galactopyranosyl residues, but 4-O of α-d-galactopyranosyl residues were branched by terminal β-d-glucopyranosyl residues. Polysaccharide C of MW 5.3 × 10⁴ composed of a chain of 1,5-linked α-d-arabinofuranosyl, 1,4-linked β-d-galactopyranosyl, 1,5-linked β-d-xylopyranosyl, 1,4-linked β-d-mannopyranosyl, 1-linked β-d-glucopyranosyl residues, and the terminal β-d-glucopyranosyl residues are attached to 3-O positions of 1,6-linked α-d-glucopyranosyl residues.     

Protein tyrosine phosphatase 1B (PTP1B) inhibitory activity and glucosidase inhibitory activity of compounds isolated from Agrimonia pilosa

Context: Despite phytochemical studies of Agrimonia pilosa Ledeb. (Rosaceae), the antidiabetic effects of this plant are unknown.

Objective: This study characterizes the isolated compounds from the aerial parts of A. pilosa and evaluates their PTP1B and α-glucosidase inhibitory properties.

Materials and methods: Ethanol extract of A. pilosa was found to inhibit 64% PTP1B activity at 30?μg/mL. The ethanol extract was partitioned with methylene chloride, ethyl acetate, n-butanol, and water fractions. Among these, the ethyl acetate fraction displayed the most potent PTP1B activity. The ethyl acetate extract was separated by chromatographic methods to obtain flavonoids and triterpenoids (1–11); which were evaluated for their inhibitory effects on PTP1B activity with p-nitrophenyl phosphate (p-NPP) as a substrate, and also α-glucosidase enzyme.

Results: Compounds 1–11 were identified as apigenin-7-O-β-d-glucuronide-6″-methyl ester, triliroside, quercetin-7-O-β-d-glycoside, quercetin-3-O-β-d-glycoside, kaempferol, kaempferol-3-O-α-l-rhamnoside, β-sitosterol, ursolic acid, tormentic acid, methyl 2-hydroxyl tricosanoate, and palmitic acid. Compounds 8, 9, and 11 displayed inhibitory effects on PTP1B activity with IC₅₀ values of 3.47?±?0.02, 0.50?±?0.06, and 0.10?±?0.03?μM, respectively. Compounds 3, 4, 6, and 9 exhibited inhibition of the α-glucosidase activity with IC₅₀ values of 11.2?±?0.2, 29.6?±?0.9, 28.5?±?0.1, and 23.8?±?0.4?μM, respectively.

Discussion and conclusion: As major ingredients of A. pilosa, compounds 1, 6, 8, and 9 showed the greatest inhibitory potency on PTP1B activity. Compounds 3, 6, 8, and 9 also showed potent inhibitory effects on α-glucosidase enzyme. This result suggested the potential of these compounds for developing antidiabetic agents.     

A new triterpenoid saponin from Ardisia gigantifolia

A new triterpenoid saponin, named 3-O-β-d-glucopyranosyl-(1 → 3)-β-d-xylopyranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 3)]-β-d-glucopyranosyl-(1 → 4)-[β-d-glucopyranosyl-(1 → 2)]-α-l-arabinopyranosyl-3β,16α,28,30-tetrahydroxy-olean-12-ene (1), along with four known triterpenoids (2–5), was isolated from the rhizomes of Ardisia gigantifolia. Their structures were elucidated by spectroscopic methods. Compounds 1–4 showed cytotoxic activity against Hela, EJ, BCG, and HepG-2 cell lines. The percentage of early apoptotic cells after treatment with 1 was significantly increased compared with control cells (p < 0.05).