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.     

Determination of the decay rate constant for hepatocytes immobilized in alginate microcapsules

Primary mouse hepatocytes (between 10–250 cells per capsule) were immobilized within 1.0% w/v alginate microbeads. The textural properties of the alginate matrix were characterized and a full protocol based upon the measurement of the initial rate of Resazurin reduction was studied and standardized. Using this method, the decay rate constant (K_d = 0.45 ± 0.01 days^?1) and the time in which the cell viability decreases in half (VI₅₀ = 37 ± 0.7 h) have been measured. The method was compared with the analysis of cell vitality using Calcein A/M and Ethidium Homodimer I. Differences between the two methods were found in the viability profile due to the significant presence of double stained cells along the culture time. According to the author's knowledge, this is the first report of a systematic study and determination of the K_d value for immobilized hepatocytes, incorporating a wide range of cell concentrations within the alginate matrix.     

Biodegradable alginate microparticles developed by electrohydrodynamic spraying techniques for oral delivery of protein

In this study, alginate microparticles were prepared by cross-linking alginate with calcium chloride solution using an electrohydrodynamic spraying technique. The effects of alginate and calcium chloride concentration as well as electrical potential on particle size and shape were investigated. The results showed that 1 mg ml^?1 alginate medium viscosity (AMV), 2.5 mg ml^?1 CaCl₂, electrical potential 18 kV (F1) and 0.5 mg ml^?1 alginate low viscosity (ALV), 2.5 mg ml^?1 CaCl₂, electrical potential 20 kV (F2) yielded the spherical shape and small particles of 937 ± 158 nm and 1556 ± 51 nm, respectively. In bovine serum albumin (BSA) entrapment efficiency study, initial BSA of 5, 10, 20, 40 and 60% w/w to polymer was incorporated into these alginate microparticles. The results revealed that F2 with initial BSA 10% w/w showed the highest entrapment efficiency of 49.70 ± 0.01%. The result of BSA content revealed that F2'with the initial BSA of 20% w/w showed the highest amount of BSA content of 3.92 ± 0.02 mg g^?1 of particles. F1 and F2 with the initial BSA of 5%, 20% and 40% w/w were chosen to evaluate for the release in PBS pH 7.4. It was found that F1 with the initial BSA of 40% w/w showed the slowest release rate and sustained release. The release of F1 in 0.1 N HCl solution (pH 1.2) was slower than that in pH 7.4. This electrohydrodynamic spray technique (EHDA) can be applied to prepare alginate in micro size and can encapsulate BSA. Alginate microparticles can further be optimized for oral delivery of several pharmaceutical peptides and proteins.     

Microencapsulation of blackberry pulp with arrowroot starch and gum arabic mixture by spray drying

Abstract

This research work aimed to obtain blackberry pulp powder by spray drying and, by an experimental design, evaluated the effect of inlet air temperature (100–150?°C) and blackberry pulp solids:arrowroot starch/gum arabic solids ratio of 1:0.5–1:2 on the physicochemical properties of the powders. Arrowroot starch and gum arabic present glass transition temperature (Tg) values above 100?°C; hence it was possible to employ them as carriers in blackberry pulp spray drying in order to increase Tg of the system. Powder yield and solubility increased with increasing blackberry pulp solids:arrowroot starch/gum arabic solids ratio of 1:0.5–1:2, whereas hygroscopicity decreased. Yield, solubility and hygroscopicity of the powders increased and water activity decreased, with increasing inlet air temperature. The powders presented low moisture content and water activity. Temperature of 143?°C and blackberry pulp solids:arrowroot starch/gum arabic solids ratio of 1:1.78 were the optimal conditions to obtain high yield and blackberry powders that are soluble in water and less hygroscopic.     

Quantitative analysis and toxicity determination of artifacts originated in a Thai traditional medicine Prasaplai

Prasaplai is a Thai traditional medicine for relieving dysmenorrhea and adjusting the menstrual cycle. Three fatty acid esters, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-yl linoleate (1), (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-yl oleate (2) and (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-yl palmitate (3) are formed during storage from the reaction of chemical components in two herbs, i.e., fatty acids in Nigella sativa (L.) (Ranunculaceae) and (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol (compound D) in Zingiber cassumunar (Roxb.) (Zingiberaceae). The formations of these artifacts were monitored for 1 year and their amounts were analyzed by HPLC at certain periods of time. The results showed that artifact formation was saturated after 73 days of storage. The amount of each artifact in the saturation period ranged from 3.93?±?0.06 to 4.30?±?0.18% w/w for compound 1, 1.69?±?0.08 to 1.9?±?0.13% w/w for compound 2 and 0.09?±?0.003 to 0.1?±?0.005% w/w for compound 3. Cytotoxicity of the artifacts was evaluated using NCI-H187, KB, and BC cancer cell lines and found that the IC₅₀ of all artifacts in all tests were higher than 20 μg/mL. For acute toxicity in mice, the LD₅₀ of each artifact was more than 300?mg/kg.     

Comparison of emulsion and vibration nozzle methods for microencapsulation of laccase and glucose oxidase by interfacial reticulation of poly(ethyleneimine)

Microcapsules for enzyme immobilization were successfully fabricated via interfacial cross-linking of poly(ethyleneimine) (PEI). A method based on laminar jet break-up technique using a commercial instrument developed to produce alginate beads is reported for the first time for production of PEI microcapsules. The diameter, wall thickness and pore size of membranes were obtained from confocal laser scanning microscopy by labelling PEI and proteins. The composition of membranes was analysed by elemental analysis. Larger microcapsules (ca 200?µm diameter) were obtained with the encapsulation device. In comparison, the emulsion method produced smaller capsules (ca 20?µm diameter) but with a wider size distribution. Encapsulation efficiency for both methods was analysed by bicinchoninic acid and fluorescence assays, yielding efficiencies of 94?±?2% and 83?±?3% for the emulsion method and encapsulation device, respectively. Glucose oxidase from Aspergillus Niger and Laccase from Trametes Versicolor were encapsulated by both microencapsulation methods and their activities were compared.     

PEG-PLGA- hybrid nanoparticles loaded with etoricoxib – phospholipid complex for effective treatment of inflammation in rat model

The aim of the present study was to increase the bioavailability of the etoricoxib by making PEG-PLGA-Hybrid nanoparticles using emulsion solvent evaporation method. Then the prepared nanoparticles were further characterised using TEM, particle size, PDI, zeta potential, encapsulation efficiency and drug release study. Lipid (Phospholipon 90-G) and drug thermal behaviour were studied using DSC, TGA. The results of optimised formulation of Particle size, PDI and zeta potential was found 216.6?±?4.0?nm, 0.24?±?0.19 and +36.3?±?1.9?mV. Encapsulation efficiency was found in the range of 77.15% w/v to 93.88% w/v. In-vivo study shows that the optimised formulation at a particular dose decreases the swelling index and number of writhes. Stability study indicated that the nanoparticles can be stored at a temperature of 4?±?2?°C/60?±?5% RH in well-closed container, away from heat and damp places. The prepared formulation has significantly increased the bioavailability of etoricoxib via oral administration.     

Amelioration of hyperglycaemia and modulation of antioxidant status by Alcea rosea seeds in alloxan-induced diabetic rats

Context: Alcea rosea L. (Malvaceae) has various medicinal uses including anticancer, anti-inflammatory and analgesic properties. However, there is no report on its antidiabetic activity.

Objective: Alcea rosea seed extracts were evaluated for antihyperglycaemic and antioxidative potential in diabetic rats.

Materials and methods: Single intra-peritoneal injection of alloxan (130?mg/kg b.w.) was used for induction of diabetes in Albino Wistar rats. Antihyperglycaemic and antioxidant activities of methanol and aqueous extracts of Alcea rosea seed (100 and 300?mg/kg b.w.), administered orally on daily basis for 15 days, were assessed in vivo for fasting blood glucose level and antioxidant status of liver and pancreas. Metformin was used as a positive control.

Results: Aqueous and methanol extracts (300?mg/kg b.w.) decreased blood glucose level in diabetic rats by 24% and 46%, respectively. Administration of aqueous and methanol extracts at 300?mg/kg b.w. significantly (p?2O₂ decomposed/min/mg of protein), respectively. Similar results were observed for pancreas.

Discussion and conclusions: Antihyperglycaemic and antioxidative potentials of Alcea rosea seeds suggest its usefulness in management of diabetes and its complications. This is the first report on antidiabetic activity of this plant.     

Development and characterization of colon specific drug delivery system bearing 5-ASA and camylofine dihydrochloride for the treatment of ulcerative colitis

The treatment of ulcerative colitis (inflammatory bowel disease, IBD) has been achieved by using colon specific drug delivery system bearing 5-ASA and Camylofine dihydrochloride. Chitosan microspheres were prepared separately for both the drugs using emulsion method followed by enteric coating with Eudragit^®S-100. The in vitro drug release was investigated in different simulated GIT medium. The drug release in PBS (pH7.4) and simulated gastric fluid has shown almost similar pattern and rate, whereas a significant increase in drug release (70.3?±?1.36 and 72.5?±?1.33% of 5-ASA and Camylofine, respectively) was observed in medium containing 3% rat caecal matter, after 24?h. In control study, 57.1?±?1.13% of 5-ASA and 59.2?±?1.2% of Camylofine release was observed in 24?h. For enzyme induction, rats were orally administered with 1?mL of 1% w/v dispersion of chitosan for 5 days and release rate studies were conducted in SCF with 3% w/v of caecal matter. An enhanced drug release (i.e., 92.3?±?3.81 and 95.5?±?3.52% 5-ASA and Camylofine, respectively) was observed after 24?h in dissolution medium containing 3% caecal content obtained from enzyme induced animals. In vivo data showed that microspheres delivered most of its drug load (76.55?±?2.13%) to the colon after 9?h, which reflects its targeting potential to the colon. It is concluded that orally administered microspheres of both drugs can be used together for the specific delivery of drug to the colon and reduce symptoms of ulcerative colitis.     

The effects of icariin on the expression of HIF-1α, HSP-60 and HSP-70 in PC12 cells suffered from oxygen–glucose deprivation-induced injury

Context: The effects of icariin, a chief constituent of ?avonoids from Epimedium brevicornum Maxim (Berberidaceae), on the levels of HIF-1α, HSP-60 and HSP-70 remain unknown.

Objective: To explore the effects of icariin on the levels of HSP-60, HIF-1α and HSP-70 neuron-specific enolase (NSE) and cell viability.

Materials and methods: PC12 cells were treated with icariin (10^?7, 10^?6 or 10^?5?mol/L) for 3?h (1?h before oxygen–glucose deprivation (OGD) plus 2?h OGD). HSP-60, HIF-1α, HSP-70 and NSE were measured using enzyme-linked immunosorbent assay (ELISA). Cell viability was determined by metabolic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

Results: After 2?h OGD, levels of HIF-1α, HSP-60, HSP-70 and NSE were increased significantly (HIF-1α: 33.3?±?1.9?ng/L, HSP-60: 199?±?16?ng/L, HSP-70: 195?±?17?ng/L, NSE: 1487?±?125?ng/L), and cell viability was significantly decreased (0.26?±?0.03), while icariin (10^?7, 10^?6, or 10^?5?mol/L) significantly reduced the contents of HIF-1α, HSP-60, HSP-70 and NSE (HIF-1α: 14.1?±?1.4, 22.6?±?1.8, 15.7?±?2.1, HSP-60: 100?±?12, 89?±?6, 113?±?11, HSP-70: 139?±?9, 118?±?7, 95?±?9 and NSE: 1121?±?80, 1019?±?52, 731?±?88), and improved cell viability (0.36?±?0.03, 0.38?±?0.04, 0.37?±?0.03) in OGD-treated PC12 cells.

Discussion and conclusion: These results indicate that the protective mechanisms of icariin against OGD-induced injury may be related to down-regulating the expression of HIF-1α, HSP-60 and HSP-70.     

GC-MS analysis and hepatoprotective activity of the n-hexane extract of Acrocarpus fraxinifolius leaves against paracetamol-induced hepatotoxicity in male albino rats

Context: In Egypt, the burden of liver diseases is exceptionally high.

Objective: To investigate the components of the n-hexane extract of Acrocarpus fraxinifolius Arn. (Leguminosae) and its hepatoprotective activity against paracetamol (APAP)-induced hepatotoxicity in rats.

Material and methods: TRACE GC ultra gas chromatogaphic spectrometry was used for extract analysis. Thirty albino rats were divided into six groups (five rats in each). Group 1 was the healthy control; Groups 2 and 3 were healthy treated groups (250 and 500?mg/kg b.w. of the extract, respectively) for seven days. Group 4 was hepatotoxicity control (APAP intoxicated group). Groups 5 and 6 received APAP?+?extract 250 and APAP?+?extract 500, respectively.

Results: Chromatographic analysis revealed the presence of 36 components. Major compounds were α-tocopherol (18.23%), labda-8 (20)-13-dien-15-oic acid (13.15%), lupeol (11.93%), phytol (10.95%) and squalene (7.19%). In the acute oral toxicity study, the mortality rates and behavioural signs of toxicity were zero in all groups (doses from 0 to 5?g/kg b.w. of A. fraxinifolius). LD₅₀ was found to be greater than 5?g/kg of the extract. Only the high dose (500?mg/kg b.w.) of extract significantly alleviated the liver relative weight (4.01?±?0.06) and biomarkers, as serum aspartate aminotransferase (62.87?±?1.41), alanine aminotransferase (46.74?±?1.45), alkaline phosphatase (65.96?±?0.74), lipid profiles (180.39?±?3.51), bilirubin profiles (2.30?±?0.06) and hepatic lipid peroxidation (114.20?±?2.06), and increased body weight (11.58?±?0.20), serum protein profile (11.09?±?0.46) and hepatic total antioxidant capacity (23.78?±?0.66) in APAP-induced hepatotoxicity in rats.

Conclusion: Our study proves the antihepatotoxic/antioxidant efficacies of A. fraxinifolius hexane extract.     

Effect of shikonin on multidrug resistance in HepG2: The role of SIRT1

Abstract

Context: Overexpression of SIRT1 is considered to enhance the resistance of HepG2 cells to irradiation. Shikonin, a naturally occurring naphthoquinone compound, displays anticancer effects and circumvents cancer drug resistance.

Objectives: This study investigated the MDR reversal effect of shikonin induced by the overexpression of SIRT1.

Materials and methods: The overexpression of SIRT1 in HepG2 cells was established by lentivirus infection. Five days after transduction, real-time quantitative polymerase chain reaction and western blotting were used to detect the expression of SIRT1 and MDR1/P-gp. Drug resistance was also evaluated by flow cytometry after rhodamine-123 staining. On day 5, the multidrug resistance cells were treated by shikonin (10^?7, 10^?6, and 10^?5 µmol/L) one time. The cell viability was detected by the MTT assay, and apoptosis was evaluated by Hoechst 33342 staining and caspase-3 activity 24?h after shikonin treatment.

Results: Overexpression of SIRT1 decreased rhodamine-123 staining and successfully produced the R-HepG2 cell line. Compared with HepG2, the expression of MDR1/P-gp mRNA (3.45?±?0.35) and protein (1.40?±?0.05) were both upregulated in R-HepG2. Shikonin inhibited cell viability (from 93.9?±?2.1 to 66.7?±?1.5%), induced apoptosis of R-HepG2 (apoptotic ratio from 3.5?±?0.8 to 47.5?±?2.7%, caspase-3 activity from 103.5?±?1.9 to 329.2?±?14.9%, respectively), downregulated the mRNA and protein expression of SIRT1 and MDR1/P-gp, and decreased rhodamin 123 efflux.

Discussion and conclusion: In the present study, we demonstrated that shikonin is able to overcome drug resistance in hepatocellular carcinoma cells, and the mechanism is related to the SIRT1-MDR1/P-gp signaling pathway.     

Pharmacokinetic and bioequivalence study of endogenous compound tretinoin 10 mg capsules in healthy volunteers by base line correction approach

Tretinoin belongs to the class of retinoids indicated in induction of remission in acute promyelocytic leukemia (APL-FAB classification AML-M3). It is an endogenous metabolite of Vitamin A and is normally present in human plasma. The objective of the present study was to evaluate the bioequivalence between the Tretinoin 10?mg capsules of Ranbaxy Laboratories Limited and VESANOID^® 10?mg capsules of Roche Pharmaceuticals Inc. It was a 2-way cross-over single dose study in 60 adult Indian male volunteers under fasting conditions. Since tretinoin is endogenously present in the human body, for baseline adjustment four pre-dose samples were collected. Plasma samples were analyzed for tretinoin by using validated liquid chromatographic mass spectrometry (LC-MS/MS) method. This method has separated both isomeric metabolites (i.e. isotretinoin and 9-oxo retinoic acid) from tretinoin to accurately measure the tretinoin concentrations in plasma for this study. The Mean ± SD of pharmacokinetic parameters T_max, C_max, and AUC_0?t for tretinoin were 2.55?±?0.84 and 2.40?±?0.86?h, 34.29?±?10.45 and 32.77?±?9.16?ng/ml, 87.28?±?30.99 and 80.81?±?24.68?ng.h/ml, respectively, for test and reference. The ratios of least square means and its 90% confidence interval for C_max, AUC_0?t and AUC_0–∞ on both baseline corrected and uncorrected data were found to be within 80–125%.     

Anti-rheumatic activity of Ananas comosus fruit peel extract in a complete Freund’s adjuvant rat model

Context: Rheumatoid arthritis is a chronic, autoimmune and systemic inflammatory disease, which targets synovial joints leading to joint destruction mediated in part by migration of inflammatory cells into the synovial tissue.

Objective: The present study evaluates the anti-rheumatic effect of a methanol extract of Ananas comosus (L.) Merr. (Bromeliaceae) peel in rats.

Materials and methods: Anti-rheumatic activity of crude extract of peels of A. comosus in complete Freund’s induced arthritis model in rats was studied at doses of 50, 100, 250 and 500?mg/kg b.w. for 21 days. Parameters such as paw size, levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), C-reactive proteins (CRP) and prostaglandins (PGE₂) were analysed.

Results: Oral administration of the extract significantly reduced the swelling in the paw of rats (EC₅₀ 65.1?±?2.95?mg/kg b.w.) with a maximal inhibition of 77.01?±?10.53% on 21st day at 500?mg/kg b.w. The extract also significantly reduced the levels of SOD, CAT and GPx in liver (EC₅₀ 26.84?±?16.37, 68.37?±?19.22, 106.54?±?34.81?mg/kg b.w., respectively), kidney (EC₅₀ 261.75?±?81.5, 176.38?±?8.08, 14.32?±?6.64, mg/kg b.w., respectively) and spleen (EC₅₀ 152.14?±?39.57, 83.97?±?14.6, 47.1?±?10.45?mg/kg b.w., respectively); and CRP (EC₅₀ 36.37?±?12.4?mg/kg b.w.) and PGE₂ (EC₅₀ 191.06?±?71.54?mg/kg b.w.) in tissue homogenate and serum, respectively, at 500?mg/kg b.w. as compared to arthritic control group.

Discussion and conclusion: These results suggest that A. comosus fruit peel extract exerts anti-rheumatic activity.     

Effectiveness of a new tobramycin (0.3 % ) and dexamethasone (0.05 % ) formulation in the treatment of experimental Pseudomonas keratitis

ABSTRACT

Objective: To quantitatively determine, in a Pseudomonas keratitis model, the anti-inflammatory and bactericidal properties of a new formulation of tobramycin (0.3?%?) and dexamethasone (0.05?%?) that utilizes a xanthan gum vehicle.

Research methods: In a randomized and masked fashion, rabbit corneas (n?≥?16 eyes per group) were intrastromally injected with 10³ colony-forming units (CFU) of P. aeruginosa. Eyes were untreated or were administered a single drop every 15?min between 16 and 17?h postinfection (PI) and then a single drop every 30?min between 17 and 22?h PI, a total of 15 drops of either 0.1?%?dexamethasone and 0.3?%?tobramycin (TobraDex; Tdex) or a new formulation 0.3?%?tobramycin and 0.05?%?dexamethasone with xanthan gum (TobraDex ST; ST). Slit lamp examination scores (SLE?±?SEM) were derived from grading seven parameters at 22?h PI. Rabbits were sacrificed at 23?h PI and the log CFU?±?SEM per cornea was determined.

Results: Untreated eyes had SLE scores of 11.11?±?0.43 and had log CFU of 7.27?±?0.06. Eyes treated with Tdex, as compared to the untreated eyes, had significantly lower SLE scores (7.39?±?0.21, p?<?0.0001) and significantly fewer bacteria (6.32?±?0.29 log CFU, p?=?0.0213). Eyes treated with ST had a SLE score (6.56?±?0.19) that was significantly lower than both the untreated eyes (p?<?0.0001) and the eyes treated with Tdex (p?=?0.0124). Furthermore, eyes treated with ST had significantly fewer log CFU (5.78?±?0.30) than untreated eyes (p?=?0.0001) or eyes treated with Tdex (p?=?0.0434).

Conclusions: The ST formulation with xanthan gum demonstrated statistically superior anti-inflammatory and bactericidal properties as compared to Tdex.

Limitations: Variations in inoculation procedures produced limited eye-to-eye differences in the infection.     

Anti-endotoxin effects of terpenoids fraction from Hygrophila auriculata in lipopolysaccharide-induced septic shock in rats

Context: Hygrophila auriculata (K. Schum) Heine (Acanthaceae) has been traditionally used for the treatment of various ailments such as inflammation, rheumatism, jaundice and malaria.

Objective: The present study aims to separate terpenoid fraction (TF) from alcohol (70%) extract of the whole plant of Hygrophila auriculata and assess its anti-inflammatory activity.

Materials and methods: HPTLC analysis of TF was performed for the estimation of lupeol. Edema was induced in Wistar albino rats by subplanter injection of 0.1?ml of 1% (w/v) carrageenan into the right hind paw after 1?h of TF administration (100 and 200?mg/kg oral). Septic shock was induced by intraperitoneal administration of LPS (100?μg/kg) in rats and interleukins (IL-1β and IL-6), tumor necrosis factor (TNF-α), superoxide dismutase (SOD), lipid peroxidation (LPO), and nitric oxide (NO) were measured in serum. AutoDock 4.2 was used for molecular docking.

Results: Administration of TF significantly (p?<?0.005) restored the serum levels of cytokines, LPO (7.77?±?0.034 versus 4.59?±?0.059?nmole of TBARS), NO (9.72?±?0.18 versus 4.15?±?0.23?µmol nitrite/mg of wet tissue), and SOD (4.89?±?0.036 versus 7.83?±?0.033?Unit/mg protein) compared with the LPS-challenged rats. Analysis of in silico results revealed that TNF-α is the most appropriate target in eliciting anti-inflammatory activity.

Conclusion: The present findings suggest that TF of Hygrophila auriculata possesses great promise as an anti-inflammatory agent which may be due to its antioxidant effect. Molecular docking results could be exploited for lead optimization and development of suitable treatment of inflammatory disorders.     

Influence of mefenamic acid on the intestinal absorption and metabolism of three bioactive flavones in Radix Scutellariae and potential pharmacological impact

Context: Mefenamic acid (MEF) and the dried root of Scutellaria baicalensis Georgi (Radix Scutellariae, RS) share a high possibility of combined medication to treat inflammation.

Objective: The present study investigates the impact of MEF on absorption/disposition of three major components in RS (baicalein, B; wogonin, W; oroxylin A, OA) and further pharmacological changes.

Materials and methods: The apparent permeability (P_app) and percentage of metabolism of B, W and OA at 10?μΜ were measured at the absence/presence of MEF (100?μΜ) in the Caco-2 cell monolayer model. A modified whole blood assay was employed to quantify prostaglandin E₂ (PGE₂) 4, 6 and 8?h post-oral administration with water suspension of MEF at 40?mg/kg and RS at 200?mg/kg.

Results: In the presence of MEF, P_app of B, W and OA were increased from 1.69?±?0.89?×?10^?6, 1.57?±?0.10?×?10^?6 and 3.09?±?0.70?×?10^?6?cm/sec to 5.24?±?0.27?×?10^?6, 6.08?±?0.19?×?10^?6 and 4.13?±?0.38?×?10^?6, whereas their percentage of metabolism was decreased from 72.75?±?2.44%, 73.27?±?3.25% and 89.84?±?2.99% to 21.11?±?0.69%, 17.90?±?5.55% and 45.44?±?3.38%. PGE₂ level was much lower in the co-administration group (49.04?±?2.03?pg/ml) than in the MEF group (73.13?±?3.03?pg/ml) or RS group (494.37?±?11.75?pg/ml) 4?h post MEF dosing, suggesting a synergic effect.

Discussion and conclusion: Co-administration of MEF and RS could induce potential alterations in their pharmacokinetic profiles and anti-inflammatory effects.     

Antiobesity and antihyperglycaemic effects of Adiantum capillus-veneris extracts: in vitro and in vivo evaluations

Context: Adiantum capillus-veneris L. (Adiantaceae) hypocholesterolemic activity is therapeutically praised.

Objectives: Pharmacological modulation of pancreatic triacylglycerol lipase (PL) and α-amylase/α-glucosidase by A. capillus-veneris are evaluated.

Materials and methods: Using positive controls (acarbose, orlistat, guar gum, atorvastatin, glipizide and metformin) as appropriate, crude aqueous extracts (AEs) of A. capillus-veneris aerial parts were tested via a combination of in vitro enzymatic (0.24–100?mg/mL), acute in vivo carbohydrate tolerance tests (125, 250 or 500?mg/kg body weight [b.wt]) and chronic in vivo studies (500?mg/kg b.wt) in high cholesterol diet (HCD) fed Wistar rats.

Results: Like acarbose, A. capillus-veneris as well as chlorogenic acid, with respective IC₅₀ values (mg/mL) of 0.8?±?0.0 and 0.2?±?0.0, were identified as in vitro potent dual inhibitors of α-amylase/α-glucosidase. Unlike guar gum, A. capillus-veneris had no glucose diffusion hindrance capacity. Equivalent to orlistat, A. capillus-veneris and its phytoconstituents inhibited PL in vitro with an ascending order of PL- IC₅₀ values (μg/mL): ferulic acid; 0.48?±?0.06?A. capillus-veneris; 1600?±?100. Incomparable to acarbose or metformin and glipizide, A. capillus-veneris (125, 250 and 500?mg/kg b.wt) lacked antihyperglycaemic efficacies in acute starch- or glucose-evoked postprandial hyperglycaemia increments in normoglycaemic overnight fasting rats. Superior to atorvastatin; A. capillus-veneris exerted significant antiobesity (p?p?Discussion and conclusion: A. capillus-veneris, modulating pancreatic digestive enzymes, may be advocated as a combinatorial diabesity prevention/phytotherapy agent.     

Potent enhancement of transdermal absorption and stability of human tyrosinase plasmid (pAH7/Tyr) by Tat peptide and an entrapment in elastic cationic niosomes

Abstract

Enhancement of transdermal absorption through rat skin and stability of the human tyrosinase plasmid (P) using Tat (T) and an entrapment in elastic cationic niosomes (E) were described. E (Tween61:cholesterol:DDAB at 1:1:0.5 molar ratio) were prepared by the freeze-dried empty liposomes (FDELs) method using 25% ethanol. TP was prepared by a simple mixing method. TPE was prepared by loading T and P in E at the T:P:E ratio of 0.5:1:160 w/w/w. For gel formulations, P, TP, PE and TPE were incorporated into Carbopol 980 gel (30?µg of plasmid per 1?g of gel). For the transdermal absorption studies, the highest cumulative amounts and fluxes of the plasmid in viable epidermis and dermis (VED) were observed from the TPE of 0.31?±?0.04?µg/cm² and 1.86?±?0.24?µg/cm²/h (TPE solution); and 4.29?±?0.40?µg/cm² and 25.73?±?2.40?µg/cm²/h (TPE gel), respectively. Only plasmid from the PE and TPE could be found in the receiving solution with the cumulative amounts and fluxes at 6?h of 0.07?±?0.01?µg/cm² and 0.40?±?0.08?µg/cm²/h (PE solution); 0.10?±?0.01?µg/cm² and 0.60?±?0.06?µg/cm²/h (TPE solution); 0.88?±?0.03?µg/cm² and 5.30?±?0.15?µg/cm²/h (PE gel); and 1.02?±?0.05?µg/cm² and 6.13?±?0.28?µg/cm²/h (TPE gel), respectively. In stability studies, the plasmid still remained at 4?±?2?°C and 25?±?2?°C of about 48.00–65.20% and 27.40–51.10% (solution); and 12.34–38.31% and 8.63–36.10% (gel), respectively, whereas at 45?±?2?°C, almost all the plasmid was degraded. These studies indicated the high potential application of Tat and an entrapment in elastic cationic niosomes for the development of transdermal gene delivery system.     

Investigation of the role of alginate containing high guluronic acid on osteogenic differentiation capacity of human umbilical cord Wharton’s jelly mesenchymal stem cells

Background and aim: Cell encapsulation using biodegradable material has promising results for tissue engineering. Since pressure is an effective factor on stem cell behaviour and various concentrations of alginate create different pressures on the cells, therefore our goal was to evaluate the mechanical effect of 1/2% (w/v) and 1/8% (w/v) alginate containing high guluronic acid on viability and osteogenic capacity of HUCWJ cells.

Methods: Cell viability was evaluated by MTT assay after 1, 7 and 14 days. Alkaline phosphatase activity was evaluated by alkaline phosphatase assay kit after 14 and 21 days. Alizarin red S staining was performed for calcium deposition among histological section.

Results: MTT assay showed significant difference in the mean of viability rates between groups in day 14 (p 相似文献