scholarly journals PTPN2 improves implant osseointegration in T2DM via inducing the dephosphorylation of ERK

2019 ◽  
Vol 244 (16) ◽  
pp. 1493-1503 ◽  
Author(s):  
Ya-Nan Wang ◽  
Tingting Jia ◽  
Jiajia Zhang ◽  
Jing Lan ◽  
Dongjiao Zhang ◽  
...  
Keyword(s):  
Osteogenic Differentiation ◽  
High Glucose ◽  
Histological Evaluation ◽  
Glucose Medium ◽  
Alizarin Red ◽  
Positive Effects ◽  
Pull Out ◽  
High Glucose Medium ◽  
Novel Target

Type 2 diabetes mellitus (T2DM) is considered to compromise implant osseointegration. Protein tyrosine phosphatase non-receptor type 2 (PTPN2) regulates glucose metabolism, systemic inflammation, and bone regeneration. This study aimed to investigate the role of PTPN2 in implant osseointegration in T2DM and explore the potential mechanisms. Streptozotocin-induced diabetic rats received implant surgery, with or without local overexpression of PTPN2 for three months, and implant osseointegration was examined by histological evaluation, micro-CT analysis, pull-out test, and scanning electron microscope. Rat bone marrow stem cells (RBMSCs) were isolated and exposed to high glucose, and osteogenic differentiation was evaluated by alizarin red staining, ALP assay, and Western blot analysis. Overexpression of PTPN2 could improve impaired implant osseointegration in T2DM rats and promote osteogenic differentiation of RBMSCs in high glucose. In addition, p-ERK level in RBMSCs was increased in high glucose and decreased after PTPN2 overexpression. These results suggest that PTPN2 promotes implant osseointegration in T2DM rats and enhances osteogenesis of RBMSCs in high glucose medium via inducing the dephosphorylation of ERK. PTPN2 may be a novel target for the therapy of impaired implant osseointegration in T2DM patients. Impact statement Using both in vivo and in vitro approaches, we made important findings that PTPN2 promoted implant osseointegration in T2DM rats and enhanced osteogenesis of RBMSCs in high glucose medium. The positive effects of PTPN2 on osteogenesis are related to the dephosphorylation of ERK and the inhibition of MAPK/ERK pathway. PTPN2 may be a novel target for the therapy of impaired implant osseointegration in T2DM patients.

scholarly journals Glucose regulates tissue-specific chondro-osteogenic differentiation of human cartilage endplate stem cells via O-GlcNAcylation of Sox9 and Runx2

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Chao Sun ◽  
Weiren Lan ◽  
Bin Li ◽  
Rui Zuo ◽  
Hui Xing ◽  
...  
Keyword(s):  
Western Blot ◽  
Osteogenic Differentiation ◽  
High Glucose ◽  
Immunohistochemical Staining ◽  
Chondrogenic Differentiation ◽  
Glucose Medium ◽  
Cartilage Endplate ◽  
High Glucose Medium ◽  
Differentiation Medium ◽  
Low Glucose

Abstract Background The degenerative disc disease (DDD) is a major cause of low back pain. The physiological low-glucose microenvironment of the cartilage endplate (CEP) is disrupted in DDD. Glucose influences protein O-GlcNAcylation via the hexosamine biosynthetic pathway (HBP), which is the key to stem cell fate. Thiamet-G is an inhibitor of O-GlcNAcase for accumulating O-GlcNAcylated proteins while 6-diazo-5-oxo-l-norleucine (DON) inhibits HBP. Mechanisms of DDD are incompletely understood but include CEP degeneration and calcification. We aimed to identify the molecular mechanisms of glucose in CEP calcification in DDD. Methods We assessed normal and degenerated CEP tissues from patients, and the effects of chondrogenesis and osteogenesis of the CEP were determined by western blot and immunohistochemical staining. Cartilage endplate stem cells (CESCs) were induced with low-, normal-, and high-glucose medium for 21 days, and chondrogenic and osteogenic differentiations were measured by Q-PCR, western blot, and immunohistochemical staining. CESCs were induced with low-glucose and high-glucose medium with or without Thiamet-G or DON for 21 days, and chondrogenic and osteogenic differentiations were measured by Q-PCR, western blot, and immunohistochemical staining. Sox9 and Runx2 O-GlcNAcylation were measured by immunofluorescence. The effects of O-GlcNAcylation on the downstream genes of Sox9 and Runx2 were determined by Q-PCR and western blot. Results Degenerated CEPs from DDD patients lost chondrogenesis, acquired osteogenesis, and had higher protein O-GlcNAcylation level compared to normal CEPs from LVF patients. CESC chondrogenic differentiation gradually decreased while osteogenic differentiation gradually increased from low- to high-glucose differentiation medium. Furthermore, Thiamet-G promoted CESC osteogenic differentiation and inhibited chondrogenic differentiation in low-glucose differentiation medium; however, DON acted opposite role in high-glucose differentiation medium. Interestingly, we found that Sox9 and Runx2 were O-GlcNAcylated in differentiated CESCs. Finally, O-GlcNAcylation of Sox9 and Runx2 decreased chondrogenesis and increased osteogenesis in CESCs. Conclusions Our findings demonstrate the effect of glucose concentration on regulating the chondrogenic and osteogenic differentiation potential of CESCs and provide insight into the mechanism of how glucose concentration regulates Sox9 and Runx2 O-GlcNAcylation to affect the differentiation of CESCs, which may represent a target for CEP degeneration therapy.

scholarly journals Alpha-Lipoic Acid Alleviates High-Glucose Suppressed Osteogenic Differentiation of MC3T3-E1 Cells via Antioxidant Effect and PI3K/Akt Signaling Pathway

2017 ◽  
Vol 42 (5) ◽  
pp. 1897-1906 ◽  
Author(s):  
Kai Dong ◽  
Pengjie Hao ◽  
Sheng Xu ◽  
Shutai Liu ◽  
Wenjuan Zhou ◽  
...  
Keyword(s):  
Oxidative Damage ◽  
Osteogenic Differentiation ◽  
Lipoic Acid ◽  
High Glucose ◽  
Osteoblastic Differentiation ◽  
Antioxidant Effect ◽  
Akt Pathway ◽  
Alpha Lipoic Acid ◽  
Western Blots ◽  
Alizarin Red

Background/Aims: Patients with diabetes mellitus have a higher risk of dental implant failure. One major cause is high-glucose induced oxidative stress. Alpha-lipoic acid (ALA), a naturally occurring compound and dietary supplement, has been established as a potent antioxidant that is a strong scavenger of free radicals. However, few studies have yet investigated the effect of ALA on osteogenic differentiation of osteoblasts cultured with high glucose medium. The aim of this study is to investigate the effects of ALA on the osteoblastic differentiation in MC3T3-E1 cells under high glucose condition. Methods: MC3T3-E1 cells were divided into 4 groups including normal glucose (5.5 mM) group (control), high glucose (25.5 mM) group, high glucose + 0.1 mM ALA group, and high glucose + 0.2 mM ALA group. The proliferation, osteogenic differentiation and mineralization of cells were evaluated by MTT assay, alkaline phosphatase (ALP) activity assay, alizarin red staining and real time-polymerase chain reaction. High-glucose induced oxidative damage was also assessed by the production of reactive oxygen species (ROS) and superoxide dismutase (SOD). Western blots were performed to examine the role of PI3K/Akt pathway. Results: The proliferation, osteogenic differentiation and mineralization of MC3T3-E1 cells were significantly decreased by the ROS induced by high-glucose. All observed oxidative damage and osteogenic dysfunction induced were inhibited by ALA. Moreover, the PI3K/Akt pathway was activated by ALA. Conclusions: We demonstrate that ALA may attenuate high-glucose mediated MC3T3-E1 cells dysfunction through antioxidant effect and modulation of PI3K/Akt pathway.

Nutrient Utilization by Bovine Articular Chondrocytes: A Combined Experimental and Theoretical Approach

2005 ◽  
Vol 127 (5) ◽  
pp. 758-766 ◽  
Author(s):  
Bram G. Sengers ◽  
Hannah K. Heywood ◽  
David A. Lee ◽  
Cees W. J. Oomens ◽  
Dan L. Bader
Keyword(s):  
Oxygen Uptake ◽  
High Glucose ◽  
Articular Chondrocytes ◽  
Lactate Production ◽  
Glucose Medium ◽  
Numerical Predictions ◽  
High Glucose Medium ◽  
Biosensor System ◽  
Wide Range ◽  
Bovine Articular Chondrocytes

A combined experimental-numerical approach was adopted to characterize glucose and oxygen uptake and lactate production by bovine articular chondrocytes in a model system. For a wide range of cell concentrations, cells in agarose were supplemented with either low or high glucose medium. During an initial culture phase of 48h, oxygen was monitored noninvasively using a biosensor system. Glucose and lactate were determined by medium sampling. In order to quantify glucose and oxygen uptake, a finite element approach was adopted to describe diffusion and uptake in the experimental model. Numerical predictions of lactate, based on simple relations for cell metabolism, were found to agree well for low glucose, but not for high glucose medium. Oxygen did not play a role in either case. Given the close association between chondrocyte energy metabolism and matrix synthesis, a quantifiable prediction of utilization can present a valuable contribution in the optimization of tissue engineering conditions.

Effect of Jinmaitong (筋脉通) serum on the proliferation of rat Schwann cells cultured in high glucose medium

2008 ◽  
Vol 14 (4) ◽  
pp. 293-297 ◽  
Author(s):  
Ling Qu ◽  
Xiao-chun Liang ◽  
Hong Zhang ◽  
Qun-li Wu ◽  
Lian-qing Sun ◽  
...  
Keyword(s):  
Schwann Cells ◽  
High Glucose ◽  
Glucose Medium ◽  
High Glucose Medium ◽  
Cells Cultured

Mitochondrial stress causes increased succination of proteins in adipocytes in response to glucotoxicity

2012 ◽  
Vol 445 (2) ◽  
pp. 247-254 ◽  
Author(s):  
Norma Frizzell ◽  
Sonia A. Thomas ◽  
James A. Carson ◽  
John W. Baynes
Keyword(s):  
High Glucose ◽  
Addition Reaction ◽  
Tricarboxylic Acid Cycle ◽  
Michael Addition Reaction ◽  
3T3 Cells ◽  
Mitochondrial Stress ◽  
High Glucose Medium ◽  
Transport Chain ◽  
Type 2 Diabetic

2SC [S-(2-succino)-cysteine] is a chemical modification formed by a Michael addition reaction of fumarate with cysteine residues in proteins. Formation of 2SC, termed ‘succination’ of proteins, increases in adipocytes grown in high-glucose medium and in adipose tissues of Type 2 diabetic mice. However, the metabolic mechanisms leading to increased fumarate and succination of protein in the adipocyte are unknown. Treatment of 3T3 cells with high glucose (30 mM compared with 5 mM) caused a significant increase in cellular ATP/ADP, NADH/NAD+ and Δψm (mitochondrial membrane potential). There was also a significant increase in the cellular fumarate concentration and succination of proteins, which may be attributed to the increase in NADH/NAD+ and subsequent inhibition of tricarboxylic acid cycle NAD+-dependent dehydrogenases. Chemical uncouplers, which dissipated Δψm and reduced the NADH/NAD+ ratio, also decreased the fumarate concentration and protein succination. High glucose plus metformin, an inhibitor of complex I in the electron transport chain, caused an increase in fumarate and succination of protein. Thus excess fuel supply (glucotoxicity) appears to create a pseudohypoxic environment (high NADH/NAD+ without hypoxia), which drives the increase in succination of protein. We propose that increased succination of proteins is an early marker of glucotoxicity and mitochondrial stress in adipose tissue in diabetes.

scholarly journals Anti-Adipogenic Activity of Berberine in 3T3-L1 Pre-Adipocytes Cultured in High-Glucose Medium

2016 ◽  
Vol 63 (10) ◽  
pp. 474-477
Author(s):  
Hiroshi Shinmoto ◽  
Yoshihito Yamaguchi ◽  
Manami Ito ◽  
Rena Hoshizaki ◽  
Yasunori Naganawa
Keyword(s):  
High Glucose ◽  
Glucose Medium ◽  
High Glucose Medium

scholarly journals CircRNA-DAPK1 promotes vascular cell pyroptosis in diabetes by regulating the circ-DAPK1/miR-4454/TXNIP axis.

2021 ◽  
Author(s):  
Tianchi Chen ◽  
Xiangtao Zheng ◽  
Yangyan He ◽  
Chenyang Qiu ◽  
Xiaohui Wang ◽  
...  
Keyword(s):  
High Glucose ◽  
Luciferase Reporter ◽  
Glucose Medium ◽  
Vascular Cell ◽  
Normal Medium ◽  
High Glucose Medium ◽  
Signaling Axis

Abstract Background Circular RNAs have been demonstrated to play an important role in the development of vascular diseases. However, little is known about the role of circ-021774, also named circ-DAPK1, in vascular cell pyroptosis. Methods Circ-DAPK1 was selected from circular RNA sequencing data of HUVECs treated with high glucose medium and normal medium. RT-qPCR was used to determine the expression of circ-DAPK1 in vivo and in vitro. Dual luciferase reporter assay, fluorescence in situ hybridization (FISH) and RNA immunoprecipitation (RIP) were performed to prove the interaction of circ-DAPK1, miRNA-4454 and thioredoxin-interactingprotein (TXNIP). Adeno-associated virus (AAV) was injected intravenously to establish mouse models. PI staining, western-blot and transmission electron microscopy (TEM) analyses were performed to identify the role of circ-DAPK1 in promoting pyroptosis. Results We found that circ-DAPK1 was highly expressed in high glucose medium cultured HUVECs and db/db mice. In vitro and in vivo experiments demonstrated that circ-DAPK1 knockdown decreased the number of PI+ cells, the expression of ASC, NLRP3, GSDMD-N, cleaved caspase-1, IL-18 and IL-1β. In a mechanistic study, the circ-DAPK1/miRNA-4454/TXNIP signaling axis was demonstrated to promote vascular cell pyroptosis in diabetes. Conclusions Circ-DAPK1 functions as a promoter of vascular cell pyroptosis in diabetes via the circ-DAPK1/miRNA-4454/TXNIP signaling axis.

scholarly journals High Glucose Enhances the Odonto/Osteogenic Differentiation of Stem Cells from Apical Papilla via NF-KappaB Signaling Pathway

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Yanping Wang ◽  
Yanqiu Wang ◽  
Yadie Lu ◽  
Jinhua Yu
Keyword(s):  
Stem Cells ◽  
Western Blot ◽  
Osteogenic Differentiation ◽  
Real Time ◽  
Signaling Pathway ◽  
Mtt Assay ◽  
High Glucose ◽  
Rt Pcr ◽  
Alizarin Red ◽  
Apical Papilla

Objective. The transport and metabolism of glucose are important during mammalian development. High glucose can mediate the biological characteristics of mesenchymal stem cells (MSCs). However, the role of high glucose in the odonto/osteogenic differentiation of stem cells from apical papilla (SCAPs) is unclear. Materials and Methods. SCAPs were isolated and identified in vitro. Then, SCAPs were cultured in normal α-MEM and high glucose α-MEM separately. MTT assay was applied to observe the proliferation of SCAPs. ALP activity, alizarin red staining, real-time RT-PCR, and western blot were used to detect the odonto/osteogenic capacity of SCAPs as well as the participation of NF-κB pathway. Results. SCAPs in 25mmol/L glucose group expressed the maximum proteins of RUNX2 and ALP as compared with those in 5, 10, and 15 mmol/L groups. MTT assay showed that 25 mmol/L glucose suppressed the proliferation of SCAPs. ALP assay, alizarin red staining, real-time RT-PCR, and western blot showed 25 mmol/L high glucose can obviously enhance the odonto/osteogenic capacity of SCAPs. Moreover, the NF-κB pathway was activated in 25mmol/L glucose-treated SCAPs and the odonto/osteogenic differentiation was inhibited following the inhibition of NF-κB signaling pathway. Conclusions. High glucose can enhance the odonto/osteogenic capacity of SCAPs via NF-κB pathway.

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