scholarly journals Conditioned Medium of Adipose-Derived Mesenchymal Stem Cells as a Promising Candidate to Protect High Glucose-Induced Injury in Cultured C28I2 Chondrocytes

2021 ◽  
Author(s):  
Sedighe Safari ◽  
Akram Eidi ◽  
Mehrnaz Mehrabani ◽  
Mohammad Javad Fatemi ◽  
Ali Mohammad Sharifi
Keyword(s):  
Lipid Peroxidation ◽  
Conditioned Medium ◽  
High Glucose ◽  
Caspase 3 ◽  
Thiobarbituric Acid ◽  
Heme Oxygenase 1 ◽  
Diabetic Patients ◽  
Mitochondrial Apoptosis ◽  
Paracrine Effects ◽  
Protein Levels

Purpose: The aim of this study was to evaluate the protective effect of conditioned medium derived from human adipose MSCs (CM-hADSCs) on C28I2 chondrocytes against oxidative stress and mitochondrial apoptosis induced by high glucose (HG). Methods: C28I2 cells were pre-treated with CM-hADSCs for 24 hours followed by HG exposure (75 mM) for 48 hours. MTT assay was used to assess the cell viability. Reactive oxygen species (ROS) and lipid peroxidation were determined by 2,7-dichlorofluorescein diacetate (DCFH-DA) and thiobarbituric acid reactive substances (TBARS) assays, respectively. Expressions of glutathione peroxidase 3 (GPX 3), heme oxygenase-1 (HO-1), and NAD(P)H quinone dehydrogenase 1 (NQO1) were analyzed by RT-PCR. Finally, western blot analysis was used to measure Bax, Bcl-2, cleaved caspase-3, and Nrf-2 expression at protein levels. Results: CM-hADSCs pretreatment mitigated the cytotoxic effect of HG on C28I2 viability. Treatment also markedly reduced the levels of ROS, lipid peroxidation, and augmented the expression of HO-1, NQO1, and GPx3 genes in HG-exposed group. CM-ADSCs enhanced Nrf-2 protein expression and reduced mitochondrial apoptosis through reducing Bax/Bcl-2 ratio and Caspase-3 activation. Conclusion: MSCs, probably through its paracrine effects, declined the deleterious effect of HG on chondrocytes. Hence, therapies based on MSCs secretomes appear to be a promising therapeutic approaches to prevent joint complications in diabetic patients.

scholarly journals Inhibition of DNA Repair Protein Ku70 in High-Glucose Environment Aggravates the Neurotoxicity Induced by Bupivacaine in SH-SY5Y Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Zhong-Hua Ji ◽  
Yu Zhou ◽  
Hui Wang ◽  
Shuang Li ◽  
Gen-qiang Liang ◽  
...  
Keyword(s):  
Dna Damage ◽  
High Glucose ◽  
Caspase 3 ◽  
Diabetic Patients ◽  
Human Neuroblastoma ◽  
Protein Levels ◽  
Dna Oxidative Damage ◽  
Repair Protein ◽  
Dna Repair Protein ◽  
Species Production

Bupivacaine, a common local anesthetic, causes serious nerve injury, especially in diabetic patients, as high glucose has been reported to enhance bupivacaine-induced neurotoxicity. However, the key regulator for synergism remains unknown. To our surprise, the expression of repair protein Ku70 is suppressed, while the high-glucose environment induces DNA oxidative damage in neurons. Here, we aim to investigate whether the inhibition of Ku70 by high-glucose conditions aggrandized bupivacaine-induced DNA damage. Consistent with previous results, bupivacaine induced reactive oxygen species production and upregulated Ku70 and cleaved caspase-3 expressions at both transcript and protein levels and ultimately caused nucleic acid damage and apoptosis in human neuroblastoma (SH-SY5Y) cells. High-glucose treatment inhibited the expression of Ku70 and enhanced bupivacaine-induced neurotoxicity. In contrast, the overexpression of Ku70 mitigated DNA damage and apoptosis triggered by bupivacaine and high glucose. In conclusion, our data indicated that local anesthetics may aggravate nerve toxicity in a high-glucose environment.

scholarly journals (Pro)renin receptor regulates autophagy and apoptosis in podocytes exposed to high glucose

2015 ◽  
Vol 309 (3) ◽  
pp. E302-E310 ◽  
Author(s):  
Caixia Li ◽  
Helmy M. Siragy
Keyword(s):  
Signaling Pathway ◽  
High Glucose ◽  
Caspase 3 ◽  
Mtor Signaling ◽  
Autophagic Flux ◽  
Bafilomycin A1 ◽  
Mtor Signaling Pathway ◽  
Autophagosome Formation ◽  
Podocyte Injury ◽  
Protein Levels

High glucose reduces autophagy and enhances apoptosis of podocytes. Previously, we reported that high glucose induced podocyte injury through upregulation of the (pro)renin receptor (PRR). We hypothesized that increasing PRR reduces autophagy and increases apoptosis of mouse podocytes exposed to high glucose via activation of the PI3K/Akt/mTOR signaling pathway. Mouse podocytes were cultured in normal (5 mmol/l) or high (25 mmol/l) d-glucose for 48 h. High glucose significantly increased mRNA and protein levels of PRR, phosphorylation of PI3K/Akt/mTOR, and p62. In contrast, high glucose decreased activation of UNC-51-like kinase-1 (ULK1) by phosphorylating Ser757 and protein levels of microtubule-associated protein-1 light chain 3B (LC3B)-II and Lamp-2. Bafilomycin A1 increased LC3BII and p62 accumulation in high-glucose-treated cells. High glucose reduced the autophagic flux. Confocal microscopy studies showed significant reduction in the protein level of LC3B in response to high glucose. Cyto-ID autophagy staining showed a significant decrease in autophagosome formation with high glucose. In the absence of PRR, activation of Akt with sc-79 or mTOR with MHY-1485 increased p62 accumulation. Caspase-3/7 activity and apoptosis monitored by TUNEL assay were significantly increased in podocytes treated with high glucose. PRR siRNA significantly reversed the effects of high glucose. Based on these data, we conclude that high glucose decreases autophagy and increases apoptosis in mouse podocytes through the PRR/PI3K/Akt/mTOR signaling pathway.

scholarly journals The Role of Heme Oxygenase 1 in the Protective Effect of Caloric Restriction against Diabetic Cardiomyopathy

2019 ◽  
Vol 20 (10) ◽  
pp. 2427 ◽  
Author(s):  
Maayan Waldman ◽  
Vadim Nudelman ◽  
Asher Shainberg ◽  
Romy Zemel ◽  
Ran Kornwoski ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Beneficial Effect ◽  
Heme Oxygenase ◽  
Diabetic Cardiomyopathy ◽  
High Glucose ◽  
Heme Oxygenase 1 ◽  
Ros Production ◽  
Diabetic Mice ◽  
Protein Levels

Type 2 diabetes mellitus (DM2) leads to cardiomyopathy characterized by cardiomyocyte hypertrophy, followed by mitochondrial dysfunction and interstitial fibrosis, all of which are exacerbated by angiotensin II (AT). SIRT1 and its transcriptional coactivator target PGC-1α (peroxisome proliferator-activated receptor-γ coactivator), and heme oxygenase-1 (HO-1) modulates mitochondrial biogenesis and antioxidant protection. We have previously shown the beneficial effect of caloric restriction (CR) on diabetic cardiomyopathy through intracellular signaling pathways involving the SIRT1–PGC-1α axis. In the current study, we examined the role of HO-1 in diabetic cardiomyopathy in mice subjected to CR. Methods: Cardiomyopathy was induced in obese diabetic (db/db) mice by AT infusion. Mice were either fed ad libitum or subjected to CR. In an in vitro study, the reactive oxygen species (ROS) level was determined in cardiomyocytes exposed to different glucose levels (7.5–33 mM). We examined the effects of Sn(tin)-mesoporphyrin (SnMP), which is an inhibitor of HO activity, the HO-1 inducer cobalt protoporphyrin (CoPP), and the SIRT1 inhibitor (EX-527) on diabetic cardiomyopathy. Results: Diabetic mice had low levels of HO-1 and elevated levels of the oxidative marker malondialdehyde (MDA). CR attenuated left ventricular hypertrophy (LVH), increased HO-1 levels, and decreased MDA levels. SnMP abolished the protective effects of CR and caused pronounced LVH and cardiac metabolic dysfunction represented by suppressed levels of adiponectin, SIRT1, PPARγ, PGC-1α, and increased MDA. High glucose (33 mM) increased ROS in cultured cardiomyocytes, while SnMP reduced SIRT1, PGC-1α levels, and HO activity. Similarly, SIRT1 inhibition led to a reduction in PGC-1α and HO-1 levels. CoPP increased HO-1 protein levels and activity, SIRT1, and PGC-1α levels, and decreased ROS production, suggesting a positive feedback between SIRT1 and HO-1. Conclusion: These results establish a link between SIRT1, PGC-1α, and HO-1 signaling that leads to the attenuation of ROS production and diabetic cardiomyopathy. CoPP mimicked the beneficial effect of CR, while SnMP increased oxidative stress, aggravating cardiac hypertrophy. The data suggest that increasing HO-1 levels constitutes a novel therapeutic approach to protect the diabetic heart. Brief Summary: CR attenuates cardiomyopathy, and increases HO-1, SIRT activity, and PGC-1α protein levels in diabetic mice. High glucose reduces adiponectin, SIRT1, PGC1-1α, and HO-1 levels in cardiomyocytes, resulting in oxidative stress. The pharmacological activation of HO-1 activity mimics the effect of CR, while SnMP increased oxidative stress and cardiac hypertrophy. These data suggest the critical role of HO-1 in protecting the diabetic heart.

scholarly journals Coenzyme Q10 Attenuates High Glucose-Induced Endothelial Progenitor Cell Dysfunction through AMP-Activated Protein Kinase Pathways

2016 ◽  
Vol 2016 ◽  
pp. 1-14 ◽  
Author(s):  
Hsiao-Ya Tsai ◽  
Chih-Pei Lin ◽  
Po-Hsun Huang ◽  
Szu-Yuan Li ◽  
Jia-Shiong Chen ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Coenzyme Q10 ◽  
High Glucose ◽  
Endothelial Progenitor Cell ◽  
Progenitor Cell ◽  
Heme Oxygenase 1 ◽  
Diabetic Patients ◽  
No Production ◽  
Endothelial Progenitor ◽  
Amp Activated Protein Kinase

Coenzyme Q10 (CoQ10), an antiapoptosis enzyme, is stored in the mitochondria of cells. We investigated whether CoQ10 can attenuate high glucose-induced endothelial progenitor cell (EPC) apoptosis and clarified its mechanism. EPCs were incubated with normal glucose (5 mM) or high glucose (25 mM) enviroment for 3 days, followed by treatment with CoQ10 (10 μM) for 24 hr. Cell proliferation, nitric oxide (NO) production, and JC-1 assay were examined. The specific signal pathways of AMP-activated protein kinase (AMPK), eNOS/Akt, and heme oxygenase-1 (HO-1) were also assessed. High glucose reduced EPC functional activities, including proliferation and migration. Additionally, Akt/eNOS activity and NO production were downregulated in high glucose-stimulated EPCs. Administration of CoQ10 ameliorated high glucose-induced EPC apoptosis, including downregulation of caspase 3, upregulation of Bcl-2, and increase in mitochondrial membrane potential. Furthermore, treatment with CoQ10 reduced reactive oxygen species, enhanced eNOS/Akt activity, and increased HO-1 expression in high glucose-treated EPCs. These effects were negated by administration of AMPK inhibitor. Transplantation of CoQ10-treated EPCs under high glucose conditions into ischemic hindlimbs improved blood flow recovery. CoQ10 reduced high glucose-induced EPC apoptosis and dysfunction through upregulation of eNOS, HO-1 through the AMPK pathway. Our findings provide a potential treatment strategy targeting dysfunctional EPC in diabetic patients.

scholarly journals Dehydroepiandrosterone prevents lipid peroxidation and cell growth inhibition induced by high glucose concentration in cultured rat mesangial cells

2000 ◽  
Vol 166 (2) ◽  
pp. 401-406 ◽  
Author(s):  
E Brignardello ◽  
M Gallo ◽  
M Aragno ◽  
R Manti ◽  
E Tamagno ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Cell Growth ◽  
High Glucose ◽  
Glucose Concentration ◽  
Mesangial Cells ◽  
Primary Cultures ◽  
Thiobarbituric Acid ◽  
Secretory Product ◽  
High Glucose Concentration ◽  
Physiological Concentration

The oxidative stress induced by high glucose concentration contributes to tissue damage associated with diabetes, including renal injury. Dehydroepiandrosterone (DHEA), the major secretory product of the human adrenal gland, has been shown to possess a multi-targeted antioxidant activity which is also effective against lipid peroxidation induced by high glucose. In this study we evaluated the effect of DHEA on the growth impairment which high glucose concentration induces in cultured rat mesangial cells. Primary cultures of rat mesangial cells were grown for 10 days in media containing either normal (i.e. 5.6 mmol/l) or high (i.e. 30 mmol/l) concentrations of glucose, without or with DHEA at different concentrations. The impairment of cell growth induced by high glucose was reversed by 100 nmol/l and 500 nmol/l DHEA, which had no effect on mesangial cells cultured in media containing glucose at the normal physiological concentration (5.6 mmol/l). In high-glucose cultured mesangial cells, DHEA also attenuated the lipid peroxidation, as measured by thiobarbituric acid reactive substances (TBARS) generation and 4-hydroxynonenal (HNE) concentration, and preserved the cellular content of reduced glutathione as well as the membrane Na+/K+ ATPase activity. The data further support the protective effect of DHEA against oxidative damage induced by high glucose concentrations, and bring into focus its possible effectiveness in preventing chronic complications of diabetes.

scholarly journals Paracrine effects of mesenchymal stem cells in cisplatin-induced renal injury require heme oxygenase-1

2011 ◽  
Vol 300 (1) ◽  
pp. F254-F262 ◽  
Author(s):  
Abolfazl Zarjou ◽  
Junghyun Kim ◽  
Amie M. Traylor ◽  
Paul W. Sanders ◽  
József Balla ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Growth Factor ◽  
Conditioned Medium ◽  
Heme Oxygenase ◽  
Morphological Changes ◽  
Heme Oxygenase 1 ◽  
Paracrine Effects ◽  
Angiogenic Potential ◽  
Promising Therapeutic Approach

Multipotent mesenchymal stem cells (MSC) have become a popular and promising therapeutic approach in many clinical conditions. MSC are beneficial in animal models of acute kidney injury (AKI), by mediating differentiation-independent paracrine properties, and have prompted ongoing clinical trials to evaluate the safety and efficacy of MSC. Heme oxygenase-1 (HO-1) is induced in response to stress including AKI and has important anti-apoptotic, anti-inflammatory, and proangiogenic properties in these settings. We therefore examined whether HO-1 plays a role in the beneficial effects of MSC in AKI. We isolated MSC from bone marrow of age-matched HO-1+/+ and HO-1−/− mice. Our studies indicate that while differentiation of MSC into osteo- and adipocytic lineages did not differ between cells isolated from HO-1+/+ and HO-1−/− mice, MSC from HO-1−/− mice had significantly lower angiogenic potential. Moreover, HO-1−/− MSC demonstrated reduced expression and secretion of several important growth and proangiogenic factors (stromal cell-derived factor-1, vascular endothelial growth factor-A, and hepatocyte growth factor) compared with MSC derived from HO-1+/+ mice. In addition, conditioned medium of HO-1+/+ MSC rescued functional and morphological changes associated with cisplatin-induced AKI, while the HO-1−/−-conditioned medium was ineffectual. Our studies indicate that HO-1 plays an important role in MSC-mediated protection. The results expand understanding of the renoprotective effects of MSC and may provide novel strategies to better utilize MSC in various disease models.

A Sensitive Method for the Measurement of Glycosylated Plasma Proteins Using Affinity Chromatography

1984 ◽  
Vol 21 (1) ◽  
pp. 16-21 ◽  
Author(s):  
Barry J Gould ◽  
Pauline M Hall ◽  
John G H Cook
Keyword(s):  
Plasma Proteins ◽  
Phenylboronic Acid ◽  
Colorimetric Method ◽  
Thiobarbituric Acid ◽  
Diabetic Patients ◽  
Good Precision ◽  
Protein Levels ◽  
Routine Measurement ◽  
Test Kit ◽  
Acid Gel

We describe a simple, sensitive affinity technique for the routine measurement of glycosylated plasma proteins in clinical laboratories. The commercially available phenylboronic acid gel used for the chromatography has recently been marketed as a kit for this purpose (Glycogel Test Kit, Pierce Chemical Co). The manufacturers of this kit recommend loading 200 μl neat plasma to each 1 ml gel column. This high loading is to enable the direct measurement of protein in the bound and unbound fractions at 280 nm. This loading is consistent with 10–15 mg protein being added per ml gel. Our results show that protein levels greater than 2 mg per ml gel overload the column. Therefore we used a modification of the more sensitive Bradford procedure to measure protein. The method discriminates between normals (6·29 ± 1·87%) and diabetic patients (12·62 ± 3·36%) and has good precision (CV 4–6%). The results obtained correlate with the colorimetric method using thiobarbituric acid ( r = 0·70) and with glycosylated haemoglobin ( r = 0·82).

α1 Anti-Trypsin (AAT) Mitigates Hematopoietic Injury and Enhances Bone Marrow Recovery After Total Body Irradiation (TBI)

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4142-4142
Author(s):  
Ekapun Karoopongse ◽  
Sabina Janciauskiene ◽  
Charles A. Dinarello ◽  
H. Joachim Deeg ◽  
A. Mario Q. Marcondes
Keyword(s):  
Bone Marrow ◽  
Dna Damage ◽  
Total Body Irradiation ◽  
Caspase 3 ◽  
Bone Marrow Cells ◽  
Heme Oxygenase 1 ◽  
Cytokine Storm ◽  
Protein Levels ◽  
Total Body ◽  
Marrow Cells

Abstract Abstract 4142 Background: Injury to healthy (non-target) tissues still is a major limitation of radiation therapy, at least in part related to release of cytokines, including TNFα and IL-1β, which amplify tissue damage. Cytokine release is particularly prominent in patients who receive total body irradiation (TBI) before hematopoietic cell transplantation (HCT), as interactions of allogeneic donor cells with patient tissue contribute to the resulting “cytokine storm” and the development of graft versus-host-disease (GVHD). In murine models, administration of alpha-1 anti-trypsin (AAT) in the peri-transplant period decreased GVHD incidence and mortality. AAT, a member of the serine protease inhibitor (serpin) family, is a major protective protein in the circulation and has been used successfully in the clinic for other indications. Methods: We were interested in determining the potential benefits of AAT in preventing toxicity related to the transplant conditioning regimen, specifically TBI. AAT inhibits proteinase-3 (PR3) which, among other targets, cleaves IL-32 thereby leading to activation of TNFa and enhancing the cytokine storm. Since others have also suggested that AAT, via enhanced expression of heme oxygenase-1 (HMOX-1), leads to activation of Nrf2, thereby enhancing transcription of anti-inflammatory cytokines such as IL10 and the IL-1 receptor antagonist (IL1Ra), we determined the overall shift in the cytokine milieu and cell death. Thus, we irradiated male C3H/HeN mice (n = 5 mice per group; 6–8 weeks old) with sub-lethal doses (500, 600 and 700 cGy) of TBI from a 137Cs source. AAT (300 μg/animal) was administered intra-peritoneally 1 hour before and every 48 hours after TBI for a total of 6 doses. The effect of TBI/AAT on hematopoiesis was analyzed by growing granulocyte-macrophage colony forming units (GM-CFU) from bone marrow cells, on days 3, 7 and 14 after TBI. Results: Results were compared to those with cells from albumin-treated controls. Marrows (days 3 and 7) from AAT-treated mice generated higher GM-CFU counts than those from controls (mean = 25 vs. 5 colonies per 25 × 103 cells plated after 500cGy, and 15 vs. 3 colonies per 25 × 103 cells after 600 cGy). Peripheral blood and unsorted bone marrow from AAT-treated mice showed up-regulation of HMOX-1 and Nrf2 (30 and 50 log2 increase, respectively), and enhanced transcription of IL-10 and IL-1Ra (50 and 10 log2, respectively) compared to albumin treated donors. PR3 and TNFα, in contrast, were down-regulated (5 log2 and 7 log2 decrease in comparison to albumin treated controls). The cytokine mitigating effects of AAT were accompanied by attenuation of ATM-p53-dependent DNA damage responses in bone marrow cells (determined on days 3, 7, and 14), showing a 3-fold decrease in p53 protein levels, and a 6-fold decrease in phospho-ATM in comparison to albumin treated controls (peak day 7). In addition, protein levels of caspase 3 and caspase 9 were decreased (2-fold and 5-fold, respectively; peak on day 7) in unsorted marrow cells and spleen lysates of AAT treated mice in comparison to albumin treated controls. Bone marrow histopathology revealed normo-cellularity and a decrease in cleaved caspase 3 staining in AAT-treated mice compared to albumin treated animals. Summary and conclusions: AAT treatment significantly mitigated the hematopoietic toxicity induced by sub-lethal TBI. The mechanism involves cytokine suppression, associated with attenuation of ATM-p53 mediated DNA damage response. Taken together, these data suggest that AAT treatment pre-TBI provides protection against radiation injury. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Protective Effects of Fucoxanthin on High Glucose- and 4-Hydroxynonenal (4-HNE)-Induced Injury in Human Retinal Pigment Epithelial Cells

Antioxidants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1176
Author(s):  
Yi-Fen Chiang ◽  
Hsin-Yuan Chen ◽  
Yen-Jui Chang ◽  
Yin-Hwa Shih ◽  
Tzong-Ming Shieh ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Epithelial Cells ◽  
Protective Effect ◽  
High Glucose ◽  
Retinal Pigment ◽  
Antioxidant Properties ◽  
The Body ◽  
Biologically Active ◽  
Diabetic Patients ◽  
Blood Retinal Barrier

The incidence of diabetes mellitus is increasing due to the eating and living habits of modern people. As the disease progresses, the long-term effects of diabetes can cause microvascular disease, causing dysfunction in different parts of the body, which, in turn, leads to different complications, such as diabetic neuropathy, diabetic nephropathy, and diabetic retinopathy (DR). DR is the main cause of vision loss and blindness in diabetic patients. Persistent hyperglycemia may cause damage to the retina, induce the accumulation of inflammatory factors, and destroy the blood–retinal barrier function. Fucoxanthin (Fx) is a marine carotenoid extracted from seaweed. It accounts for more than 10% of the total carotenoids in nature. Fx is mainly found in brown algae and has strong antioxidant properties, due to its unique biologically active structure. This carotenoid also has the effects of reducing lipid peroxidation, reducing DNA damage, and preventing cardiovascular diseases as well as anti-inflammatory and anti-tumor effects. However, there is no relevant research on the protective effect of Fx in DR. Therefore, in this study, we explore the protective effect of Fx on the retina. Human retinal epithelial cells (ARPE-19) are used to investigate the protective effect of Fx on high glucose stress- (glucose 75 mM) and high lipid peroxidation stress (4-hydroxynonenal, 4-HNE (30 μM))-induced DR. The cell viability test shows that Fx recovered the cell damage, and Western blotting shows that Fx reduced the inflammation response and maintained the integrity of the blood–retinal barrier by reducing its apoptosis and cell adhesion factor protein expression. Using an antioxidant enzyme assay kit, we find that the protective effect of Fx may be related to the strong antioxidant properties of Fx, which increases catalase and reduces oxidative stress to produce a protective effect on the retina.

scholarly journals Danzhi Jiangtang Capsule Mediates NIT-1 Insulinoma Cell Proliferation and Apoptosis by GLP-1/Akt Signaling Pathway

2019 ◽  
Vol 2019 ◽  
pp. 1-7 ◽  
Author(s):  
Yuan-Jie Wu ◽  
Yuan-Bo Wu ◽  
Zhao-Hui Fang ◽  
Mei-Qiao Chen ◽  
Yu-Feng Wang ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
High Glucose ◽  
Caspase 3 ◽  
Cellular Proliferation ◽  
High Dose ◽  
Glucose Load ◽  
Akt Signaling Pathway ◽  
Protein Levels ◽  
Proliferation And Apoptosis ◽  
Cellular Apoptosis

Objective. This study aimed to investigate the effects of Danzhi Jiangtang Capsule (DJC) on the proliferation and apoptosis functions of NIT-1 pancreatic β-cells exposed to high-glucose load through GLP-1 activated Akt/ FoxO1 signaling pathway. Methods. Cellular apoptosis of NIT-1 pancreatic β-cells was induced by culturing in medium with 33.3mmol/L high glucose (HG). Then low-dose DJC (HG +LD), high-dose DJC (HG +HD), high-dose DJC+ GLP-1 inhibition (HG +HD +GI), and high-dose DJC+AKT inhibition (HG +HD+AI) were added, respectively. Cellular proliferation was accessed by cell counting kit (CCK-8) and cellular apoptosis was measured by Annexin V-FITC/PI staining. The protein levels of phosphorylated phosphatidylinositol-3-kinase (p-PI3K), phosphorylated AKT (p-AKT), phosphorylated Forkhead box protein O1 (p-FoxO1), and cleaved caspase-3 were detected by Western blotting. The mRNA expression of pancreatic duodenal homeobox-1 (PDX-1), CyclinD1, Bcl-2, and insulin was tested by Q-PCR. Results. Comparing to HG group, (HG+HD) group showed a significantly increased cellular proliferation. The apoptosis of NIT-1 cells also was obviously reduced, with downregulated cleaved caspase-3 protein level and upregulated PDX-1, CyclinD1, and Bcl-2 mRNA levels (P<0.05). Additionally, (HG+HD) group manifested increased insulin mRNA expression; the protein levels of p-PI3K and p-AKT were markedly increased and p-FoxO1 was decreased. All of the above therapeutic effects by DJC intervention had been reversed by GLP-1 inhibition in (HG+HD+GI) group or AKT inhibition in (HG+HD+AI) group. Conclusion. DJC was able to attenuate the toxicity of high-glucose load in NIT-1 pancreatic β-cells, ascribed to the improvement of cellular proliferation and apoptosis by GLP-1/Akt signaling pathway. This study could supply a new mechanism of DJC effects on type 2 diabetes mellitus (T2DM) treatment.

Sign in / Sign up

Export Citation Format

Share Document