Polyphenols from Euphorbia pekinensis Inhibit AGEs Formation In Vitro and Vessel Dilation in Larval Zebrafish In Vivo

Planta Medica ◽  
2017 ◽  
Vol 84 (03) ◽  
pp. 176-181 ◽  
Author(s):  
Ik-Soo Lee ◽  
Seung-Hyun Jung ◽  
Jin Kim

AbstractTo identify active compounds in the roots of Euphorbia pekinensis for treatment of diabetic complications, an active column fraction from a 70% EtOH extract of E. pekinensis root was purified by preparative reversed-phase high-performance liquid chromatography, leading to the isolation of a new ellagic acid derivative, 3,3′-di-O-methylellagic acid 4-O-(6ʺ-O-galloyl)-β-D-galactopyranoside (1), along with three known compounds, geraniin (2), 3,3′-di-O-methylellagic acid 4-O-β-D-xylopyranoside (3), and ellagic acid 3,3′-dimethyl ether (4). The structure of the new compound was established by extensive spectroscopic studies and chemical evidence. The inhibitory effects of isolated compounds 1–4 on advanced glycation end-products (AGEs) formation were examined. All compounds exhibited considerable inhibition of AGEs formation and IC50 values of 0.41 – 12.33 µM, compared with those of the positive controls aminoguanidine (IC50 = 1122.34 µM) and quercetin (IC50 = 27.80 µM). In addition, the effects of 2 and 4 on the dilation of hyaloid-retinal vessels induced by high glucose (HG) in larval zebrafish were investigated; both compounds significantly reduced the HG-induced dilation of hyaloid-retinal vessels relative to the HG-treated control group.

Dose-Response ◽  
2020 ◽  
Vol 18 (3) ◽  
pp. 155932582093942
Author(s):  
Muhammad Younus ◽  
Muhammad Mohtasheem ul Hasan ◽  
Khalil Ahmad ◽  
Ali Sharif ◽  
Hafiz Muhammad Asif ◽  
...  

In this study, we aimed to investigate the antidiabetic effects of Euphorbia nivulia (En), native to Cholistan Desert area of Bahawalpur, Pakistan. First, we performed high-performance liquid chromatography analysis and found that this plant contains ferulic acid, gallic acid, quercetin, benzoic acid, polyphenols, and flavonoids. Then, we performed in vitro and in vivo studies to assess its effects on diabetic Wistar rat model. The experiments were performed and compared with control drug glibenclamide. The 70% hydroalcoholic extract of En exhibited 97.8% in vitro α-glucosidase inhibitory effect at a dose of 1.0 mg/mL. We orally administered the extract of En and control drug to the streptozotocin (STZ)-induced diabetic rats and analyzed its antidiabetic effects. We found that the extract of En with a dose of 500 mg/kg/body weight exhibited significant effect to reduce blood glucose in STZ-induced rats as compared with the control group ( P < .001). Our histological data also showed that the extract significantly improved the histopathology of pancreas. Collectively, both in vitro and in vivo studies revealed that En possesses α-glucosidase inhibitory, antioxidant, and anti-hyperglycemic effect in STZ-induced diabetic rats.


Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3525
Author(s):  
João E. Oliveira ◽  
Miriam F. Suzuki ◽  
Renata Damiani ◽  
Eliana R. Lima ◽  
Kleicy C. Amaral ◽  
...  

Human BMP-2, a homodimeric protein that belongs to the TGF- β family, is a recognized osteoinductor due to its capacity of inducing bone regeneration and ectopic bone formation. The administration of its recombinant form is an alternative to autologous bone grafting. A variety of E. coli-derived hBMP-2 has been synthesized through refolding of cytoplasmic inclusion bodies. The present work reports the synthesis, purification, and characterization of periplasmic hBMP-2, obtained directly in its correctly folded and authentic form, i.e., without the initial methionine typical of the cytoplasmic product that can induce undesired immunoreactivity. A bacterial expression vector was constructed including the DsbA signal peptide and the cDNA of hBMP-2. The periplasmic fluid was extracted by osmotic shock and analyzed via SDS-PAGE, Western blotting, and reversed-phase high-performance liquid chromatography (RP-HPLC). The purification was carried out by heparin affinity chromatography, followed by high-performance size-exclusion chromatography (HPSEC). HPSEC was used for qualitative and quantitative analysis of the final product, which showed >95% purity. The classical in vitro bioassay based on the induction of alkaline phosphatase activity in myoblastic murine C2C12 cells and the in vivo bioassay consisting of treating calvarial critical-size defects in rats confirmed its bioactivity, which matched the analogous literature data for hBMP-2.


1994 ◽  
Vol 141 (1) ◽  
pp. 123-129 ◽  
Author(s):  
F de Pablo ◽  
R Dashner ◽  
A R Shuldiner ◽  
J Roth

Abstract Insulin is a multifunctional polypeptide hormone that regulates metabolic processes and promotes mitogenesis and differentiation in vitro in the cells and tissues of several species. Its role in vivo during embryogenesis is still poorly understood. We have previously found insulin mRNA in mature Xenopus laevis oocytes and in embryos during neurulation (before organogenesis of the pancreas takes place). We have now measured insulin immunoactivity in mature oocytes, unfertilized eggs and day-2 tadpoles. Using reversed phase high performance liquid chromatography, we found low levels of insulin in extracts of oocytes (stage VI). Both Xenopus insulin I and II were detected in unfertilized eggs. The day-2 tadpoles (stages 31–33) also contained immunoactive insulin, and in swimming tadpoles (stage 46) a few clusters of cells containing insulin immunoactivity could be identified by indirect immunofluorescence. Immunoblot analysis was relatively insensitive, detecting insulin only in the adult Xenopus pancreas. In summary, insulin (from maternal origin and embryonic expression) appears to be present early enough in Xenopus laevis to influence developmental processes such as neurulation. Journal of Endocrinology (1994) 141, 123–129


2020 ◽  
Author(s):  
Jianwu Zhang ◽  
Lan Yang ◽  
Yingying Fang ◽  
Jing Li ◽  
Chunxia Gu ◽  
...  

Abstract BackgroundOphioglossum vulgatum Linn. (Ophioglossaceae) (OV), which is traditionally used on wounds and burns, enjoys a reputation as the king of medicine in Taiwan. There are few studies on its role in gastrointestinal diseases. Our aim was to assess the antidiarrheal and spasmolytic effect of the ethanol whole plant extract of Ophioglossum vulgatum (EWOV).MethodsStudy was conducted from June 2018 to July 2019. The chemical constituents of EWOV were analyzed by high performance liquid chromatography (HPLC). In vivo, the antidiarrheal activity of EWOV (125, 250 and 500 mg/kg; orally) in castor oil-induced Kun Ming mice was evaluated. In vitro, the effect of EWOV (0.01-10 mg/mL) on the spontaneous contraction of isolated rabbit jejunum smooth muscle was studied. Verapamil was the positive control group in both vivo and vitro studies. The jejunum stripes were pre-contracted by ACh (10-5 M) and KCl (60 mM) which could induce the jejunum spasm. The possible spasmolytic effect was analyzed in the pretreatment of the jejunum preparations with EWOV (0.3, 1 mg/mL) or verapamil (0.03, 0.1 µM) in Ca2+-free and high-K+ (60 mM) solution containing EDTA.ResultsEWOV (250 and 500 mg/kg) exhibited antidiarrheal effect. EWOV (0.01-10 mg/mL) inhibited the spontaneous and ACh/KCl-induced contraction with an EC50 value of 1.46 (0.89-2.04), 1.06 (0.63-1.48) and 0.48 (0.29-0.67), and it shifted the concentration-response curves of CaCl2 to right with decreased in max, similar to verapamil. ConclusionsEWOV has significant antidiarrheal and spasmolytic effect, possibly by mediating calcium channel blocking activity, this provides the pharmacological basis for use in gastrointestinal disorders.


2003 ◽  
Vol 94 (6) ◽  
pp. 2237-2240 ◽  
Author(s):  
Sebastiano Gangemi ◽  
Graziella Luciotti ◽  
Etrusca D'Urbano ◽  
Agostino Mallamace ◽  
Domenico Santoro ◽  
...  

Lipoxins (LX) are lipoxygenase-derived eicosanoids with potent anti-inflammatory activities and vascular bed-dependent vasodilatory actions. LX can be formed in vitro and in vivo in a number of conditions, and we have reported that immunoreactive LXA4 (iLXA4) is physiologically excreted with human urine. Using a recently developed LX extraction method coupled to an ELISA, we examined whether iLXA4 excretion was modified by strenuous exercise, which is known to trigger potential LX-forming events. Maximal exertion significantly increased iLXA4 urinary excretion in nine healthy volunteers (0.061 ± 0.023 vs. 0.113 ± 0.057 ng/mg creatinine; P = 0.028). iLXA4 levels returned to baseline after 6 h and increased, although at a smaller extent, after 24 h. A significant correlation ( r = 0.988) was denoted between iLXA4 ELISA measurements and reversed-phase high-performance liquid chromatography quantitation of a previously described urinary tetraene, confirming its LXA4-related nature. These findings show for the first time that an increase in excretion of LXA4-related compounds can be observed in response to strenuous exercise. This may be the reflection of an enhanced LX biosynthesis, which may represent a safeguard mechanism that keeps the inflammatory reaction triggered by physical stress under control.


2020 ◽  
Vol 59 (1) ◽  
pp. 71-78
Author(s):  
Jaber Emami ◽  
Moloud Kazemi ◽  
Anahita Salehi

Abstract The relative in vitro and in vivo evaluation of two hydroxychloroquine (HCQ) products was conducted. In vitro studies involved assay, content uniformity and dissolution test, and a two-way crossover fashion were used for in vivo studies. Blood samples were collected at appropriate intervals and HCQ levels were measured using a validated reversed-phase high-performance liquid chromatography (HPLC) method. The drug and the internal standard, chloroquine (CQ), were extracted from blood with diethyl ether, separated and dried under nitrogen gas. Residues were reconstituted in the mobile phase and analyzed at 340 nm on a μ-bondapack C18 (250 × 4.6 mm) HPLC column with acetonitrile:methanol:KH2PO4 (10:10:80) mixture containing 0.01% triethylamine. The standard curve was linear within 50–1,500 ng/mL HCQ (R2 = 0.9996), relative errors were 1.6 to 5%, and the CV% ranged from 7 to 15.4. The resolution factor and RSD were 1.62 and 0.35% and in vitro data of both products met the USP requirements. The 90% confidence intervals for the ratios of the AUC0–96, Cmax and Tmax and their corresponding logarithmically transformed values of generic product over those of Plaquenil® were within the acceptable limit of 0.80–1.20 and 0.80–1.25, respectively. Therefore, the generic HCQ was bioequivalent to the innovator formulation.


1988 ◽  
Vol 118 (1) ◽  
pp. 119-124 ◽  
Author(s):  
Mogens Fenger ◽  
Linda Hilsted

Abstract. The effect of ascorbic acid depletion on the amidation of alphamelanocyte stimulating hormone (αMSH) was studied in vivo in guinea pig pituitary. After four weeks, the concentration of ascorbic acid was 1.20 ± 0.11 μmol/g tissue (mean ± sd) in the pituitary and 0.34 ± 0.07 μmol/g tissue in the cerebral cortex from the depleted animals versus 7.58 ± 0.08 and 1.51 ± 0.32 μmol/g tissue, respectively, in the control animals. In the pituitaries from the animals depleted of ascorbate (N = 4), the relative amount of αMSH was reduced to approximately half the values obtained in the control group (from 66.5 ± 4.6% of total ACTH-related peptides to 31.1 ± 12.2% (P < 0.0025)). A concomitant increase (from 5.9 ±3.1% to 19.4 ± 4.3% (P < 0.004)) in ACTH (1–14) (the glycine-extended precursor of αMSH) immunoreactivity and a smaller increase in ACTH (1–39) immunoreactivity was observed in the depleted guinea pigs. Gel chromatography and reversed-phase high-performance luquid chromatography showed that the αMSH and ACTH (1–14) immunoreactivity was of low molecular weight and partly mono- or diacetylated. Depletion of ascorbic acid had no influence on the degree of acetylation of αMSH and ACTH (1–14). It is concluded that depletion of ascorbic acid reduces the in vivo amidation of ACTH (1–14) in the guinea pig pituitary.


1982 ◽  
Vol 60 (5) ◽  
pp. 645-650 ◽  
Author(s):  
S. S. Gnanamanickam ◽  
A. N. Starratt ◽  
E. W. B. Ward

Coronatine was detected in culture filtrates of 12 out of 19 pathogenic strains of Pseudomonas glycinea examined, but not in culture filtrates of strains of P. phaseolicola, P. syringae, or P. tabaci. A reversed-phase high-performance liquid chromatographic procedure was developed for coronatine quantitation. Generally, coronatine production by strains in culture correlated with their ability to induce systemic symptoms (chlorosis and stunting) in inoculated soybean plants. Application of purified preparations of coronatine to unifoliate leaves of soybean plants resulted in localized chlorosis, development of chlorosis in subsequently developing trifoliate leaves, and stunting of plant growth, similar to symptoms induced by infection. Coronatine was demonstrated in soybean leaves infected with P. glycinea but was not detected in healthy leaves. The results indicate that coronatine can play an important role in the development of symptoms of bacterial blight of soybean, but the demonstration that some pathogenic strains do not produce coronatine indicates that it may not be essential for pathogenicity.


1990 ◽  
Vol 29 (03) ◽  
pp. 120-124
Author(s):  
R. P. Baum ◽  
E. Rohrbach ◽  
G. Hör ◽  
B. Kornhuber ◽  
E. Busse

The effect of triiodothyronine (T3) on the differentiation of cultured neuroblastoma (NB) cells was studied after 9 days of treatment with a dose of 10-4 M/106 cells per day. Using phase contrast microscopy, 30-50% of NB cells showed formation of neurites as a morphological sign of cellular differentiation. The initial rise of the mitosis rate was followed by a plateau. Changes in cyclic nucleotide content, in the triphosphates and in the activity of the enzyme ornithine decarboxylase (ODC) were assessed in 2 human and 2 murine cell lines to serve as biochemical parameters of the cell differentiation induced by T3. Whereas the cAMP level increased significantly (3 to 7 fold compared with its initial value), the cGMP value dropped to 30 to 50% of that of the control group. ATP and GTP increased about 200%, the ODC showed a decrease of about 50%. The present studies show a biphasic effect of T3 on neuroblastoma cells: the initial rise of mitotic activity is followed by increased cell differentiation starting from day 4 of the treatment.


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