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Shuyun Liu, Lanlan Zhang, Jingqiu Cheng, Yanrong Lu, Jingping Liu
Inflammatory response is a major cause of grafts dysfunction in islet transplantation. Hepatocyte growth factor (HGF) had shown anti-inflammatory activity in multiple diseases. In this study, we aim to deliver HGF by self-assembling peptide/heparin (SAP/Hep) hybrid gel to protect β-cell from inflammatory injury. The morphological and slow release properties of SAPs were analyzed. Rat INS-1 β-cell line was treated with tumor necrosis factor α in vitro and transplanted into rat kidney capsule in vivo, and the viability, apoptosis, function, and inflammation of β-cells were evaluated...
2016: International Journal of Nanomedicine
Adam Neal, Austin Rountree, Kelly Kernan, Brian Van Yserloo, Huiliang Zhang, Benjamin J Reed, William Osborne, Wang Wang, Ian R Sweet
A real time method to measure intracellular H2O2 would be very impactful in characterizing rapid changes that occur in physiologic and pathophysiologic states. Current methods do not provide the sensitivity, specificity and spatiotemporal resolution needed for such experiments on intact cells. We developed the use of HyPer, a genetic indicator for H2O2 that can be expressed in the cytosol (cyto-HyPer) or the mitochondria (mito-HyPer) of live cells. INS-1 cells or islets were permeabilized and the cytosolic HyPer signal was a linear function of extracellular H2O2, allowing fluorescent cyto-HyPer signals to be converted to H2O2 concentrations...
October 11, 2016: Biochemical Journal
Rikard G Fred, Syrina Mehrabi, Christopher M Adams, Nils Welsh
OBJECTIVES: Insulin expression is highly controlled on the posttranscriptional level. The RNA binding proteins (RBPs) responsible for this result are still largely unknown. METHODS AND RESULTS: To identify RBPs that bind to insulin mRNA we performed mass spectrometry analysis on proteins that bound synthetic oligonucloetides mimicing the 5'- and the 3'-untranslated regions (UTRs) of rat and human insulin mRNA in vitro. We observed that the RBPs heterogeneous nuclear ribonucleoprotein (hnRNP) U, polypyrimidine tract binding protein (PTB), hnRNP L and T-cell restricted intracellular antigen 1-related protein (TIA-1-related protein; TIAR) bind to insulin mRNA sequences, and that the in vitro binding affinity of these RBPs changed when INS-1 cells were exposed to glucose, 3-isobutyl-1-methylxanthine (IBMX) or nitric oxide...
September 2016: Heliyon
Mahmoud El Azzouny, Melissa J Longacre, Israr-Ul H Ansari, Robert T Kennedy, Charles F Burant, Michael J MacDonald
OBJECTIVE: Glucose-stimulated insulin secretion in pancreatic beta cells requires metabolic signals including the generation of glucose-derived short chain acyl-CoAs in the cytosol from mitochondrially-derived metabolites. One concept of insulin secretion is that ATP citrate lyase generates short chain acyl-CoAs in the cytosol from mitochondrially-derived citrate. Of these, malonyl-CoA, is believed to be an important signal in insulin secretion. Malonyl-CoA is also a precursor for lipids...
October 2016: Molecular Metabolism
A Bowen, K Kos, J Whatmore, S Richardson, H J Welters
The Wnt signalling pathway in beta-cells has been linked to the development of type 2 diabetes. Investigating the impact of a non-canonical Wnt ligand, Wnt4, on beta-cell function we found that in INS-1 cells, Wnt4 was able to completely block Wnt3a stimulated cell growth and insulin secretion. However, despite high levels of Wnt4 protein being detected in INS-1 cells, reducing the expression of Wnt4 had no impact on cell growth or Wnt3a signalling. As such, the role of the endogenously expressed Wnt4 in beta-cells is unclear, but the data showing that Wnt4 can act as a negative regulator of canonical Wnt signalling in beta-cells suggests that this pathway could be a potential target for modulating beta-cell function...
October 28, 2016: Biochemical and Biophysical Research Communications
Hongxi Sun, Yu Li, Bei Sun, Ningning Hou, Juhong Yang, Miaoyan Zheng, Jie Xu, Jingyu Wang, Yi Zhang, Xianwei Zeng, Chunyan Shan, Bai Chang, Liming Chen, Baocheng Chang
BACKROUND: Type 2 diabetes has become a global epidemic disease. Atorvastatin has become a cornerstone in the prevention and treatment of atherosclerosis. However, increasing evidence showed that statins can dose-dependently increase the risk of diabetes mellitus. The mechanism is not clear. OBJECTIVE: The Ras complex pathway (Ras/Raf/extracellular signal-regulated kinase [ERK]/cAMP response element-binding protein [CREB]) is the major pathway that regulates the gene transcription...
September 2016: Medicine (Baltimore)
Y T Liu, S R Li, Z Wang, J Z Xiao
Objective: To profile the gene expression changes associated with endoplasmic reticulum stress in INS-1-3 cells induced by thapsigargin (TG) and tunicamycin (TM). Methods: Normal cultured INS-1-3 cells were used as a control. TG and TM were used to induce endoplasmic reticulum stress in INS-1-3 cells. Digital gene expression profiling technique was used to detect differentially expressed gene. The changes of gene expression were detected by expression pattern clustering analysis, gene ontology (GO) function and pathway enrichment analysis...
September 13, 2016: Zhonghua Yi Xue za Zhi [Chinese medical journal]
Xin Liu, Yun-Pan Li, Zhen-Min Zhong, Hui-Qi Tan, Hao-Peng Lin, Shao-Jun Chen, Yu-Cai Fu, Wen-Can Xu, Chi-Ju Wei
The objective of this study was to formulate a novel gene delivery system based on the erythrocyte ghost (EG) integrated with fusogenic viral glycoprotein vesicular stomatitis virus glycoprotein G (VSV-G). VSV-G proteins were harvested as condition medium of Ad293 cells carrying a VSV-G transgene and then incorporated into EG. Plasmid DNA was condensed by various transfection reagents. A luciferase expression construct (pGL3-control) and a DsRed expression cassette (pCMV-DsRed) were used to evaluate the delivery efficiency of DNA/EG/VSV-G complexes...
September 24, 2016: Applied Biochemistry and Biotechnology
Jun Sung Moon, Udayakumar Karunakaran, Suma Elumalai, In-Kyu Lee, Hyoung Woo Lee, Yong-Woon Kim, Kyu Chang Won
AIM/HYPOTHESIS: Cluster determinant 36 (CD36), a fatty acid transporter, was reported to have a pivotal role in glucotoxicity-induced beta cell dysfunction. However, little is known about how glucotoxicity influences CD36 expression, and it is unknown whether this action can be counteracted by metformin. In the present study, we showed that metformin counteracts glucotoxicity by alleviating oxidative and endoplasmic reticulum (ER) stress-induced CD36 expression. METHODS: We used primary rat islets as well as INS-1 cells for 72h to 24h with 30mM glucose, respectively...
September 9, 2016: Journal of Diabetes and its Complications
Pontus Dunér, Israa Mohammad Al-Amily, Arvind Soni, Olof Asplund, Fateme Safi, Petter Storm, Leif Groop, Stefan Amisten, Albert Salehi
CONTEXT: Adhesion G protein-coupled receptor G1 (ADGRG1) is the most abundant GPCR in human pancreatic islets, but its role in islet function is unclear. OBJECTIVE: Investigate how ADGRG1 expression and activation by its ligand, collagen III, impacts β-cell function in normal and type-2 diabetic islets. DESIGN: Genes associated with the ADGRG1 in human islets was probed by RNA-sequencing of human pancreatic islet isolated from cadaveric donors, followed by functional studies on β-cell proliferation, apoptosis and insulin secretion in human and mouse islets and in INS-1 cells...
September 16, 2016: Journal of Clinical Endocrinology and Metabolism
Jia You, Zai Wang, Shiqing Xu, Wenjian Zhang, Qing Fang, Honglin Liu, Liang Peng, Tingting Deng, Jinning Lou
Advanced glycation end products (AGEs) are believed to be involved in diverse complications of diabetes mellitus. Overexposure to AGEs of pancreatic β-cells leads to decreased insulin secretion and cell apoptosis. Here, to understand the cytotoxicity of AGEs to pancreatic β-cells, we used INS-1-3 cells as a β-cell model to address this question, which was a subclone of INS-1 cells and exhibited high level of insulin expression and high sensitivity to glucose stimulation. Exposed to large dose of AGEs, even though more insulin was synthesized, its secretion was significantly reduced from INS-1-3 cells...
2016: Journal of Diabetes Research
Yan Yang, Jiangong Ren, Yuzhen Tong, Xuejian Hu, Qingguo Lv, Nanwei Tong
Lipoapoptosis plays an important role in the pathogenesis of type 2 diabetes. Peroxisome proliferator-activated receptor delta (PPARdelta), a vital regulator of glucose and lipid metabolism, may reduce fatty acid-induced pancreatic β cell lipotoxicity in diabetes. However, the detailed molecular mechanisms underlying this process are not fully understood. In this study, we investigated the effect of activation of PPARdelta on palmitate-induced β cell apoptosis, and we explored the potential mechanism of the antiapoptotic effect...
September 8, 2016: Lipids
Jing-Ying Pan, Shijin Yuan, Tao Yu, Cong-Lin Su, Xiao-Long Liu, Jun He, He Li
Huntingtin associated protein 1 (Hap1) was originally identified as a protein that binds to the Huntington disease protein, huntingtin. Growing evidence has shown that Hap1 participates in intracellular trafficking via its association with various microtubule- dependent transporters and organelles. Recent studies also revealed that Hap1 is involved in exocytosis such as insulin release from pancreatic beta cells.However, the mechanism underlying the action of Hap1 on insulin release remains to be investigated...
September 13, 2016: Journal of Biological Chemistry
Yanan Hao, Chunyan Liu, Fei Yin, Yonglan Zhang, Jianhui Liu
Our previous work showed that geniposide affected glucose-stimulated insulin secretion (GSIS) via regulating glucose uptake and metabolism in pancreatic β cells; however, the molecular mechanisms remain largely unknown. Substantial evidence suggests that activation of 5'-AMP-activated protein kinase (AMPK) plays a central role in GSIS. Here, we aim to determine the role of AMPK on geniposide-regulated GSIS in rat pancreatic INS-1 cells. The results demonstrated that 6-[4-(2-piperidin-1-yletoxy)-phenyl]-3-pyridin-4-yl-pyrazolo[1,5-α] pyrimidine (Compound C; an AMPK inhibitor) significantly attenuated the effects of geniposide on glucose uptake, energy metabolism, and insulin secretion in INS-1 cells...
September 7, 2016: Journal of Natural Medicines
Na Wang, De-Yu Guo, Xiong Tian, Hao-Peng Lin, Yun-Pan Li, Shao-Jun Chen, Yu-Cai Fu, Wen-Can Xu, Chi-Ju Wei
OBJECTIVES: We previously found niacin receptor GPR109A was expressed in murine islet beta-cells, and signaling through GPR109A inhibited glucose stimulated insulin secretion (GSIS). However, the expression of GPR109A in human islets and its functional relevance is still not known. METHODS: The expression of GPR109A was examined by antibody staining and in situ hybridization on pancreatic paraffin sections. GPR109A was cloned and expressed in INS-1 islet beta-cells...
October 1, 2016: General and Comparative Endocrinology
Jonatan Sjölander, Elin Byman, Klaudia Kulak, Sara C Nilsson, Enming Zhang, Ulrika Krus, Gunilla T Westermark, Petter Storm, Ben C King, Erik Renström, Anna M Blom
C4b-binding protein (C4BP) is a polymer of 7 identical alpha chains and one unique beta chain synthesized in liver and pancreas. We showed previously that C4BP enhances islet amyloid polypeptide (IAPP) fibril formation in vitro. Now we report that polymeric C4BP strongly inhibited lysis of human erythrocytes incubated with monomeric IAPP while no lysis was observed after incubation with preformed IAPP fibrils. In contrast, incubation with the monomeric α-chain of C4BP was less effective. These data indicate that polymeric C4BP with multiple binding sites for IAPP enhances polymerization and neutralization of lytic activity of IAPP...
August 26, 2016: Journal of Biological Chemistry
Michal Jurášek, Petr Džubák, Silvie Rimpelová, David Sedlák, Petr Konečný, Ivo Frydrych, Soňa Gurská, Marián Hajdúch, Kateřina Bogdanová, Milan Kolář, Tomáš Müller, Eva Kmoníčková, Tomáš Ruml, Juraj Harmatha, Pavel B Drašar
Sesquiterpene lactone trilobolide is a sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor, thus depleting the Ins(1,4,5)P3-sensitive intracellular calcium stores. Here, we describe a synthesis of a series of 6 trilobolide-steroids conjugates (estradiol, pregnene, dehydroepiandrosterone, and testosterone). We found that the newly synthesized Tb-based compounds possess different remarkable biological activities. Cancer cell cytotoxicity and preferential selectivity is represented in our study by a Tb-pregnene derivative...
August 17, 2016: Steroids
Lizbeth de la Cruz, Erika I Puente, Arturo Reyes-Vaca, Isabel Arenas, Julieta Garduño, Jorge Bravo-Martínez, David E Garcia
Phosphatidylinositol-4,5-bisphosphate (PIP2) is a membrane phosphoinositide that regulates the activity of many ion channels. Influx of calcium primarily through voltage-gated calcium (CaV) channels promotes insulin secretion in pancreatic β-cells. However, whether CaV channels are regulated by PIP2, as is the case for some non-insulin-secreting cells, is unknown. The purpose of this study was to investigate whether CaV channels are regulated by PIP2 depletion in pancreatic β-cells through activation of a muscarinic pathway induced by oxotremorine methiodide (Oxo-M)...
October 1, 2016: American Journal of Physiology. Cell Physiology
Liron Danielpur, Yang-Sung Sohn, Ola Karmi, Chen Fogel, Adar Zinger, Abdulsalam Abu-Libdeh, Tal Israeli, Yael Riahi, Orit Pappo, Ruth Birk, David H Zangen, Ron Mittler, Zvi-Ioav Cabantchik, Erol Cerasi, Rachel Nechushtai, Gil Leibowitz
CONTEXT: Type 2 Wolfram syndrome (T2-WFS) is a neuronal and β-cell degenerative disorder caused by mutations in the CISD2 gene. The mechanisms underlying β-cell dysfunction in T2-WFS are not known, and treatments that effectively improve diabetes in this context are lacking. OBJECTIVE: Unraveling the mechanisms of β-cell dysfunction in T2-WFS and the effects of treatment with GLP-1 receptor agonist (GLP-1-RA). DESIGN AND SETTING: A case report and in vitro mechanistic studies...
October 2016: Journal of Clinical Endocrinology and Metabolism
Gerhardt J Boukes, Maryna van de Venter
Pancreatic cancer is only the 12th most common cancer, but the fourth leading cause of cancer-related deaths in the world. This is due to late prognosis, poor response to chemotherapy and early metastases. Natural prodrugs may play an important role in the treatment of pancreatic cancer. The main aim of this study was to determine the cytotoxicity of five natural prodrugs, namely harpagoside, hyperoside, hypoxoside, oleuropein and polydatin, by investigating apoptosis and autophagy as possible mechanism/s of action...
July 23, 2016: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
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