Ginsenoside Rd ameliorates high glucose-induced retinal endothelial injury through AMPK-STRT1 interdependence.

Diabetic retinopathy (DR) manifests as a complicated and blinding complication in diabetes mellitus. First-line treatments for advanced DR have shown ocular side-effects in some patients. Ginsenoside Rd (Rd), an active ingredient isolated from Panax notoginseng and P. ginseng, has demonstrated diverse and powerful activities on neuroprotection, anticancer and anti-inflammation, but its vascular protective effects have rarely been reported. Herein, this study aims to investigate the protective effects of Rd on retinal endothelial injury with emphasis on AMPK/SIRT1 interaction. The results indicated that Rd promoted AMPK activation and SIRT1 expression. Besides, Rd strengthened the interaction between AMPK and SIRT1 by increasing NAD+/NADH levels and LKB1 deacetylation in endothelial cells. Moreover, Rd reversed high glucose-induced activation of NOX2, oxidative stress, mitochondrial dysfunction, and endothelial apoptosis in an AMPK/SIRT1-interdependent manner. Hyperglycemia induced loss of endothelial cells and other retinal damage, which was restored by Rd via activating AMPK and SIRT1 in vivo. The enhancement of AMPK/SIRT1 interaction by Rd beneficially modulated oxidative stress and apoptosis, and ameliorated diabetes-driven vascular damage. These data also supported the evidence for Rd clinical development of pharmacological interventions and provided a novel potential vascular protective drug for early DR.

Regulation and function of AP-1 in insulinoma cells and pancreatic ß-cells.

Cav1.2 L-type voltage-gated Ca2+ channels play a central role in pancreatic ß-cells by integrating extracellular signals with intracellular signaling events leading to insulin secretion and altered gene transcription. Here, we investigated the intracellular signaling pathway following stimulation of Cav1.2 Ca2+ channels and addressed the function of the transcription factor activator protein-1 (AP-1) in pancreatic ß-cells of transgenic mice. Stimulation of Cav1.2 Ca2+ channels activates AP-1 in insulinoma cells. Pharmacological and genetic experiments identified c-Jun N-terminal protein kinase as a signal transducer connecting Cav1.2 Ca2+ channel activation with gene transcription. Moreover, the basic region-leucine zipper proteins ATF2 and c-Jun or c-Jun-related proteins were involved in stimulus-transcription coupling. We addressed the functions of AP-1 in pancreatic ß-cells analyzing a newly generated transgenic mouse model. These transgenic mice expressed A-Fos, a mutant of c-Fos that attenuates DNA binding of c-Fos dimerization partners. In insulinoma cells, A-Fos completely blocked AP-1 activation following stimulation of Cav1.2 Ca2+ channels. The analysis of transgenic A-Fos-expressing mice revealed that the animals displayed impaired glucose tolerance. Thus, we show here for the first time that AP-1 controls an important function of pancreatic ß-cells in vivo, the regulation of glucose homeostasis.

Electrosynthesis of three-dimensional nanoporous nickel on screen-printed electrode used for the determination of narirutin in citrus wastewater.

In this study, an electrochemical sensor was designed for the detection of narirutin using three-dimensional nanostructured porous nickel on screen-printed electrode (3DnpNi/SPE). The modified electrode was successfully synthesized by the dynamic hydrogen bubble template method. The 3DnpNi/SPE was characterized by spectroscopic, microscopic, and electrochemical methods. The results showed that the 3DnpNi/SPE presents good electrocatalytic activity for the oxidation of narirutin. The quantification of narirutin was conducted by differential pulse voltammetry, which showed a wide concentration range (1.0 × 10-7 - 1.0 × 10-5 mol L-1), with low detection limit (3.9 × 10-8 mol L-1), and excellent sensitivity (0.31 A L mol-1). The proposed electrode was applied toward the determination of narirutin in yellow water sample from the citrus industry, where it presented a good degree of accuracy. The 3DnpNi/SPE showed repeatability, long-term stability, and selectivity. The results obtained showed agreement with those obtained by HPLC/DAD method. Chemical compounds studied in this article.

Evaluation of polyphenol bioaccessibility and kinetic of starch digestion of spaghetti with persimmon (Dyospyros kaki) flours coproducts during in vitro gastrointestinal digestion.

The aim was to study the in vitro starch digestibility, the free and bound polyphenol profile and their bioaccessibility and antioxidant activity during in vitro gastrointestinal digestion of durum wheat semolina spaghetti added with two types of persimmon flour concentrates ("Rojo Brillante" flour and "Triumph" flour) at two concentrations (3 and 6%). Results obtained showed that persimmon flour improves the polyphenol profile of spaghetti by addition gallic acid and coumaric acid-o-hexoside, and increasing 2-fold and around 3-fold its content in spaghetti with 3% and 6% persimmon flours, respectively. Cooked process and digestion affected more to free polyphenol content than bound. Furthermore, 3% persimmon flour enriched spaghetti reduce kinetic of starch digestion, while 6% enriched spaghetti increased it. In conclusion, persimmon flours (Rojo Brillante and Triumph) at low concentrations could be used to develop spaghetti with more polyphenol content and less starch digestibility than traditional spaghetti.

Ternary complex factor regulates pancreatic islet size and blood glucose homeostasis in transgenic mice.

A hallmark of diabetes mellitus is the inability of pancreatic ß-cells to secrete sufficient amounts of insulin for maintaining normoglycemia. The formation of smaller islets may underlie the development of a diabetic phenotype, as a decreased ß-cell mass will produce an insufficient amount of insulin. For a pharmacological intervention it is crucial to identify the proteins determining ß-cell mass. Here, we identified the ternary complex factor (TCF) Elk-1 as a regulator of the size of pancreatic islets. Elk-1 mediates, together with a dimer of the serum-response factor (SRF), serum response element-regulated gene transcription. Elk-1 is activated in glucose-treated pancreatic ß-cells but the biological functions of this protein in ß-cells are so far unknown. Elk-1 and homologous TCF proteins are expressed in islets and insulinoma cells. Gene targeting experiments revealed that the TCF proteins show redundant activities. To solve the problem of functional redundancy of these homologous proteins, we generated conditional transgenic mice expressing a dominant-negative mutant of Elk-1 in pancreatic ß-cells. The mutant competes with the wild-type TCFs for DNA and SRF-binding. Expression of the Elk-1 mutant in pancreatic ß-cells resulted in the generation of significantly smaller islets and increased caspase-3 activity, indicating that apoptosis was responsible for the reduction of the pancreatic islet size. Glucose tolerance tests revealed that transgenic mice expressing the dominant-negative mutant of Elk-1 in pancreatic ß-cells displayed impaired glucose tolerance. Thus, we show here for the first time that TCF controls important functions of pancreatic ß-cells in vivo. Elk-1 may be considered as a new therapeutic target for the treatment of diabetes.