scholarly journals Moderate-intensity exercise alleviates pyroptosis by promoting autophagy in osteoarthritis via the P2X7/AMPK/mTOR axis

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Zihao Li ◽  
Ziyu Huang ◽  
He Zhang ◽  
Jinghan Lu ◽  
Yicheng Tian ◽  
...  
Keyword(s):  
Knee Joint ◽  
Signaling Pathway ◽  
Exercise Group ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Moderate Intensity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mtor Signaling Pathway ◽  
Moderate Intensity Exercise ◽  
Chondrocyte Death

AbstractInstability and excessive use of the knee joint can cause osteoarthritis (OA). Reasonable exercise can enhance the stability of the knee joint and prevent and relieve the occurrence and development of OA. As a key switch for inflammation, P2X purinoceptor 7 (P2X7) has attracted much attention in studies of OA. Exercise can regulate P2X7 expression and activation. However, the role of P2X7 in exercise-based prevention and treatment of OA is unknown. We previously showed that moderate-intensity exercise can significantly alleviate OA symptoms. Accordingly, in this study, we evaluated the effects of exercise on P2X7 expression and activation in chondrocytes. Micro-computed tomography, hematoxylin, and eosin staining, Toluidine Blue O staining, immunohistochemistry, and terminal deoxynucleotidyl transferase dUTP nick-end labeling experiments showed that P2X7 expression was lower in the moderate-intensity exercise group than in the inflammation and low- and high-intensity exercise groups. Additionally, chondrocyte death, cartilage destruction, and the degree and severity of pyroptosis were significantly reduced, whereas autophagy levels were significantly increased in the moderate-intensity exercise group. Cell Counting Kit-8 assay, lactate dehydrogenase release, flow cytometry, enzyme-linked immunosorbent assay, cell fluorescence, western blot, reverse transcription-quantitative polymerase chain reaction, and transmission electron microscopy experiments showed that moderate activation of P2X7 promoted autophagy through the AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway and promoted autolysosome targeting for degradation of the inflammasome component NLRP3, thereby inhibiting pyroptosis. Additionally, the use of AMPK and mTOR activators and inhibitors indicated that the AMPK-mTOR signaling pathway, as the downstream of P2X7, played a key role in delaying the occurrence and development of OA. We propose that moderate-intensity exercise promoted chondrocyte autophagy through the P2X7/AMPK/mTOR signal axis to alleviate pyroptosis. Our findings provide novel insights into the positive and preventative effects of exercise on OA.

Kaempferol reversals retinal ischemia/reperfusion (IR) injury through activating of PI3K/Akt/mTOR signaling pathway

2021 ◽  
Author(s):  
Da Sun ◽  
Fusheng Shang ◽  
Dagui Chen ◽  
Wenwen Wang ◽  
lili lin
Keyword(s):  
Oxidative Damage ◽  
Signaling Pathway ◽  
Inflammatory Cytokines ◽  
Ischemia Reperfusion ◽  
Retinal Ischemia ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Mtor Signaling Pathway ◽  
Autophagy Gene ◽  
Retinal Vascular Occlusion

Abstract Purpose Retinal ischemia/reperfusion (IR) injury is associated with many ocular diseases, including acute glaucoma, diabetic retinopathy, and retinal vascular occlusion. However, currently there are no effective medications to prevent the development ofretinal IR injury.Kaempferol is a kind of plant extract which has showed an excellent ability to inhibit the inflammation.. Materials and Methods In this study, both in vitro and in vivo retinaloxidative damage models were established.Cell viability was assessed by Cell Counting Kit-8 assay. Apoptosis was examined using flow cytometry analysis.Atherosclerotic lesion analysis was performed using hematoxylin-eosin staining,The expressions of Inflammatory cytokines were detected by quantitative real-time PCR and ELISA respectively.The effect of expression of apoptosis、utophagy and the PI3K/Akt/mTOR signaling pathway related pathway was evaluated by Western blot. Results We found kaempferol was able to protect the viability of ARPE-19 cells against oxidative damage by reducing its apoptosis. In addition, it also kept structurally complete epithelium, stroma and endothelium of cornea after oxidative damage. Moreover, it also able to reduce the expression of inflammatory cytokines and increased the expression of anti-inflammatory cytokines.Kaempferol was able to enhanced the expression of anti-apoptotic genes BCL-2, and reduced the expression of autophagy gene Beclin 1 and increased the expression of anti-autophagy gene LC-3,was also able to enhance the expression PI3K and the phosphorylation ofAkt andmTOR. Conclusion Kaempferolreversals retinal ischemia/reperfusion (IR) injury through activating of PI3K/Akt/mTOR signaling pathway.

scholarly journals Critical Roles of the PI3K-Akt-mTOR Signaling Pathway in Apoptosis and Autophagy of Astrocytes Induced by Methamphetamine

2019 ◽  
Vol 17 (1) ◽  
pp. 96-104 ◽  
Author(s):  
Han-Qing Liu ◽  
Ya-Wen An ◽  
A-Zhen Hu ◽  
Ming-Hua Li ◽  
Jue-Lian Wu ◽  
...  
Keyword(s):  
Western Blot ◽  
Signaling Pathway ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Critical Role ◽  
Mammalian Target Of Rapamycin ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Mtor Signaling Pathway ◽  
Induced Apoptosis

AbstractThis study aimed to reveal potential roles of the phosphatidylinositol 3 kinase (PI3K)-protein kinase B (Akt)-mammalian target of rapamycin (mTOR) signaling pathway in apoptosis and autophagy of astrocytes induced by methamphetamine (METH). A Cell Counting Kit-8 (CCK-8) was used to determine the reduction in proliferation of U-118 MG cells induced by METH. Hoechst 33258 and flow cytometry were used to observe the astrocytes. Western blot analysis was performed to evaluate protein expression and phosphorylation levels. METH inhibited the proliferation of U-118 MG cells and induced apoptosis and autophagy. Western blot analysis showed that the ratio of LC3-II/I was increased, whereas the expression of Bcl-2 was decreased. The phosphorylation cascade of kinases in the PI3K-Akt-mTOR signaling pathway was significantly inhibited by METH exposure, as were proteins downstream of mTORC1, such as p70s6k, rps6, 4EBP1 and eIF4E. METH inhibited proliferation of U-118 MG cells and induced apoptosis and autophagy. The PI3K-Akt-mTOR signaling pathway likely plays a critical role in these effects.

scholarly journals Metformin-Induced MicroRNA-34a-3p Downregulation Alleviates Senescence in Human Dental Pulp Stem Cells by Targeting CAB39 through the AMPK/mTOR Signaling Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Shuo Zhang ◽  
Rong Zhang ◽  
Pengyan Qiao ◽  
Xiaocao Ma ◽  
Rongjian Lu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Signaling Pathway ◽  
Calcium Binding ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Mtor Signaling Pathway ◽  
Dental Tissues

Dental pulp stem cells (DPSCs) are ideal seed cells for the regeneration of dental tissues. However, DPSC senescence restricts its clinical applications. Metformin (Met), a common prescription drug for type 2 diabetes, is thought to influence the aging process. This study is aimed at determining the effects of metformin on DPSC senescence. Young and aging DPSCs were isolated from freshly extracted human teeth. Flow cytometry confirmed that DPSCs expressed characteristic surface antigen markers of mesenchymal stem cells (MSCs). Cell Counting Kit-8 (CCK-8) assay showed that a concentration of 100 μM metformin produced the highest increase in the proliferation of DPSCs. Metformin inhibited senescence in DPSCs as evidenced by senescence-associated β-galactosidase (SA-β-gal) staining and the expression levels of senescence-associated proteins. Additionally, metformin significantly suppressed microRNA-34a-3p (miR-34a-3p) expression, elevated calcium-binding protein 39 (CAB39) expression, and activated the AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. Dual-luciferase reporter assay confirmed that CAB39 is a direct target for miR-34a-3p. Furthermore, transfection of miR-34a-3p mimics promoted the senescence of DPSCs, while metformin treatment or Lenti-CAB39 transfection inhibited cellular senescence. In conclusion, these results indicated that metformin could alleviate the senescence of DPSCs by downregulating miR-34a-3p and upregulating CAB39 through the AMPK/mTOR signaling pathway. This study elucidates on the inhibitory effect of metformin on DPSC senescence and its potential as a therapeutic target for senescence treatment.

scholarly journals Rapamycin Antagonizes BCRP-Mediated Drug Resistance Through the PI3K/Akt/mTOR Signaling Pathway in mPRα-Positive Breast Cancer

2021 ◽  
Vol 11 ◽  
Author(s):  
Jing Zhang ◽  
Jing Hu ◽  
Weiwei Li ◽  
Chunyan Zhang ◽  
Peng Su ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Signaling Pathway ◽  
Therapeutic Option ◽  
Poor Response ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Mtor Signaling Pathway ◽  
Potential Biomarker ◽  
Response To Chemotherapy

PurposeOverexpression of breast cancer (BCa) resistance protein (BCRP) is detected in approximately 30% of BCa cases. BCRP indicates a poor response to chemotherapy, and it has become a classic target to overcome drug-resistant tumor cells. In this study, we aimed to explore the mechanism of BCRP overexpression and a strategy to reverse this overexpression in invasive BCa.MethodsBCRP expression in BCa tissues was determined by immunohistochemistry. GSE25066 was downloaded from the NCBI GEO database. Western blot was used to determine the expression of key molecules in vitro. Cell counting kit-8 assays were used to assess the drug response of BCa cells.ResultsOur results suggested that BCRP is an independent risk factor for BCa. We further established that upon 17α-PG binding, membrane progesterone receptor α (mPRα) promoted BCRP expression via the PI3K/Akt/mTOR signaling pathway. mPRα physically interacted with p-Akt1 S473. Moreover, rapamycin, an inhibitor of mTOR complex 1 (mTORC1), downregulated BCRP expression and enhanced the effects of particular drugs, including doxorubicin and paclitaxel.ConclusionBCRP is a potential biomarker of poor prognosis in BCa. BCRP expression is regulated by 17α-PG in mPRα-positive BCa cells through the PI3K/Akt/mTOR signaling pathway. Rapamycin might enhance the therapeutic effect of chemotherapy agents in mPRα-positive MDA-MB-453/BCRP cells and might be a therapeutic option for mPRα-positive invasive BCa with BCRP overexpression.

scholarly journals LncRNA Hoxb3os protects podocytes from high glucose-induced cell injury through autophagy dependent on the Akt-mTOR signaling pathway

2021 ◽  
Author(s):  
Juan Jin ◽  
Jianguang Gong ◽  
Li Zhao ◽  
Yiwen Li ◽  
Qiang He
Keyword(s):  
Diabetic Nephropathy ◽  
Signaling Pathway ◽  
High Glucose ◽  
Cell Injury ◽  
Damage Model ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Tunel Staining ◽  
Mtor Signaling Pathway ◽  
Potential Biomarker

Background: Diabetic nephropathy (DN) is in the first place of the causes that lead to end-stage renal disease in the world. Thus, it is urgent to develop a novel diagnostic or therapeutic strategy that could stop the progression of diabetic nephropathy. Methods: RNA-sequencing was conducted in high glucose (HG)-treated MPC5 cells (podocytes). Cell morphology was examined under a light microscope. Upon high-glucose challenge, the effects of lncRNA Hoxb3os overexpression on MPC5 cells apoptosis, viability, autophagy and Akt-mTOR signaling were evaluated using flow cytometry, Cell Counting Kit-8, qRT-PCR, and Western blotting. TUNEL staining and ELISA were performed to confirm the establishment of DN model in db/db mice. Results: High-glucose exposure dramatically altered lncRNA expression profile in MPC5 cells (fold change>2), including 305 upregulated lncRNAs and 451 downregulated lncRNAs. LncRNA Hoxb3os expression was significantly reduced in the HG-induced podocyte damage model, as well as in the renal tissues from db/db mice with spontaneous DN. Overexpression of Hoxb3os significantly reduced the apoptosis rate and increased the viability of MPC5 cells under HG conditions. Further study revealed that exogenous Hoxb3os increased autophagy level in HG-exposed MPC5 cells via abrogating Akt-mTOR signaling pathway and that the process was possibly implicated in the upregulation of SIRT1. Conclusion: LncRNA Hoxb3os protected podocytes from HG-induced damage by regulating Akt-mTOR pathway and cell autophagy. Thus, lncRNA Hoxb3os appears as a potential biomarker in the diagnosis and treatment of DN in the future.

scholarly journals Winyanghuayin Decoction and Xiaoqinglong Decoction Treatment Airway Inflammation of Cold Asthma Through PI3K-mTOR Signal Pathway

2020 ◽  
Author(s):  
xing chang ◽  
Zhang Lu ◽  
Zhang Qingxiang ◽  
tian Zhang ◽  
qingyan meng ◽  
...  
Keyword(s):  
Airway Inflammation ◽  
Signaling Pathway ◽  
Lung Tissue ◽  
Protein Level ◽  
Lavage Fluid ◽  
Mtor Signaling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mtor Signaling Pathway ◽  
Better Regulation ◽  
Ice Water

Abstract Background: Airway inflammation plays a pivotal role in cold asthma. Winyanghuayin decoction and Xiaoqinglong decoction can significantly improve airway inflammation. The aim of the present study was to explore the therapeutic effect and mechanism of Winyanghuayin decoction and Xiaoqinglong decoction on OVA-induced cold asthma in rat and compare the effects of Winyanghuayin decoction and Xiaoqinglong decoction for treatment of cold asthma ratsMethods: The cold asthma rat model was induced by OVA and ice water swim stress, treated with Winyanghuayin decoction and Xiaoqinglong decoction respectively. The pulmonary function of asthmatic rats was detected by MIR Spirolab; The pathological changes of lung tissue were observed by HE staining; The autophagy in the lung of rats were observed by transmission electron microscopy; The eosinophils, lymphocytes, macrophages and total cells in bronchoalveolar lavage fluid (BALF) were counted under light microscope. The expression of IL-10\IL-13\TNF-α and TGF-β1 in serum were measured by enzyme-linked immunosorbent assay (ELISA); The protein level of LC3-II\I, ATG, PI3K and mTOR expression in lung tissues were detected by Western blot; Real-time polymerase chain reaction (RT-PCR) detected the expression of autophagy gene ATG3\ATG5\ATG7 and ATG12.Results: Winyanghuayin decoction and Xiaoqinglong decoction can alleviate airway inflammation and improve airway hyper-responsiveness, significantly increased the level of autophagy in lung tissue and decreased the protein level of PI3K and mTOR in lung tissue of cold asthma rats. Winyanghuayin decoction have better regulation ability on autophagy and PI3K-mTOR signaling pathway.Conclusion: Winyanghuayin decoction and Xiaoqinglong decoction may alleviate the airway inflammation symptoms by enhancing autophagy in lung tissue dependent on the PI3K-mTOR signaling pathway. Winyanghuayin decoction was more effective than Xiaoqinglong decoction for treatment of cold asthma.

scholarly journals Krüppel-like factor 8 promotes aerobic glycolysis in prostate cancer cells by regulating AKT/mTOR signaling pathway

2020 ◽  
Vol 19 (10) ◽  
pp. 2091-2096
Author(s):  
Cunming Zhang ◽  
Song Chen ◽  
Lide Song ◽  
Haibo Ye ◽  
Junwei Wang
Keyword(s):  
Prostate Cancer ◽  
Cell Viability ◽  
Signaling Pathway ◽  
Therapeutic Target ◽  
Aerobic Glycolysis ◽  
Lactic Acid Production ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Mtor Signaling Pathway

Purpose: To investigate the effects of Krüppel-like factor 8 (KLF8) in prostate cancer (PCa) cell viability and glycolysis, and explore its role as a regulatory factor.Methods: Immunoblot assays were conducted to assess the expression of KLF8 and proteins in AKT/mTOR pathway in PCa cell lines PC-3 and DU145. Cell Counting Kit-8 assays were performed to assess the effect of KLF8 on PCa cell viability. The glycolysis capacity of PCa cells was determined by measuring the levels of glucose intake, lactic acid production, and cellular ATP levels.Results: Depletion of KLF8 decreased the survival of PCa cells in vitro (p < 0.05). KLF8 depletion also inhibited aerobic glucose metabolism in PCa cells (p < 0.05). Further studies confirmed that KLF8 contributed to the growth and glycolysis of PCa cells via the regulation of AKT/mTOR pathway.Conclusion: KLF8 regulates glycolysis in PCa cells by regulating AKT/mTOR signaling pathway and is thus a promising therapeutic target for PCa treatment. Keywords: Krüppel-like factor 8 (KLF8), Prostate cancer (PCa), Aerobic glucose, AKT/mTOR signaling pathway, Therapeutic target

scholarly journals Tenacissoside H Induces Autophagy and Radiosensitivity of Hepatocellular Carcinoma Cells by PI3K/Akt/mTOR Signaling Pathway

Dose-Response ◽  
2021 ◽  
Vol 19 (2) ◽  
pp. 155932582110110
Author(s):  
Jiatian Lin ◽  
Jiyin Ruan ◽  
Hao Zhu ◽  
Zaizhong Chen ◽  
Junhui Chen ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Signaling Pathway ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Dependent Manner ◽  
Mtor Signaling Pathway ◽  
Concentration Dependent Manner ◽  
Protein Levels ◽  
Underlying Mechanisms ◽  
Hcc Cells

Tenacissoside H (TEH), which has anti-inflammatory and anti-tumor effects, is a major active ingredient extracted from the stem of Marsdenia tenacissima. However, the effect of TEH on hepatocellular carcinoma (HCC) as well as the underlying mechanisms are still indistinct. Presently, HCC cells (including Huh-7 and HepG2) were dealt with different concentrations of TEH. The proliferation and apoptosis of HCC cells were determined via Cell Counting Kit-8 (CCK8) assay and flow cytometry. In addition, Western blot was conducted to evaluate the expressions of autophagy—and apoptosis-related proteins. Tissue immunofluorescence was carried out to evaluate LC3B expression in the tumor tissues. The data showed that TEH suppressed the growth of HCC cells in a concentration-dependent manner. Besides, TEH enhanced radiosensitivity and promoted the apoptosis of HCC cells. Moreover, the mRNA and protein levels of autophagy-related genes (LC3-II/LC2-I, ATG5, Beclin-1) were significantly promoted by TEH. Mechanistically, TEH attenuated the activation of PI3K/Akt/mTOR signaling pathway. However, inhibition of PI3 K pathway abolished the anti-tumor effects of TEH in HCC cells. Collectively, this study suggested that TEH increases the radiosensitivity of HCC cells via inducing autophagy and apoptosis through downregulating PI3K/Akt/mTOR signaling pathway.

scholarly journals Acute Interval and Continuous Moderate Intensity Exercise Enhanced Circadian Thermogenic Activity through Browning related Genes in Obese Adolescent Female

2021 ◽  
Vol 17 (5) ◽  
pp. 566-581
Author(s):  
Sugiharto Sugiharto ◽  
Desiana Merawati ◽  
Adi Pranoto ◽  
Purwo Sri Rejeki ◽  
Moch. Nasmay Lupita ◽  
...  
Keyword(s):  
Exercise Group ◽  
Pharmacological Therapy ◽  
Moderate Intensity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Peroxisome Proliferator ◽  
Moderate Intensity Exercise ◽  
Oxidation Activity ◽  
Obese Adolescent ◽  
Significant Difference

Thermogenesis is associated with oxidation activity in muscle and fat tissue, the target of non-pharmacological therapy in preventing the increase in obesity. This research was designed to reveal the circadian profile of thermogenic gene expression after the acute interval and continuous moderate-intensity exercise. The subjects were 22 randomly selected obese adolescent females who met the predetermined inclusion criteria. The study subjects were then divided into three groups: control group (CG), acute interval moderate-intensity exercise group (AIMIE), and acute continuous moderate-intensity exercise group (ACMIE). Acute interval and continuous exercise were performed by running on a treadmill for 40-45 minutes, while moderate-intensity was defined as 60%-70% of the maximum heart rate (HRmax). The blood samples were collected initially (pre-exercise), followed by 10 minutes, 6 hours, and 24 hours post-acute interval and continuous moderate-intensity exercise treatment. Measurement of peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α) and fibronectin type III domain 5 (FNDC-5) expressions in protein level were confirmed by enzyme-linked immunosorbent assay (ELISA) method. Data were analyzed using one way-ANOVA and two way-ANOVA with a significant level of 5%. The findings suggest a substantial increase in the expression of PGC-1α and FNDC-5 after exercise compared to before the workout. A significant difference in PGC-1α and FNDC-5 expressions between the control group compared to AIMIE and ACMIE (p ≤ 0.05) has been observed. However, there is no significant difference in PGC-1α and FNDC-5 expressions after exercise between AIMIE and ACMIE (p ≥ 0.05). In conclusion, acute interval and continuous moderate-intensity exercise increase the expression of thermogenesis-related genes. Hence, acute interval and continuous moderate-intensity exercise might be potential non-pharmacological therapy to prevent, reduce, and control the increasing prevalence of obesity.

Effect of PI3K/AKT/mTOR Signaling Pathway on Regulating and Controlling the Anti-Invasion and Metastasis of Hepatoma Cells by Bufalin

2021 ◽  
Vol 16 ◽  
Author(s):  
Xia Sheng ◽  
Pengfei Zhu ◽  
Yi Zhao ◽  
Jinwei Zhang ◽  
Haijia Li ◽  
...  
Keyword(s):  
Protein Expression ◽  
Signaling Pathway ◽  
Inhibitory Effect ◽  
Mtor Signaling ◽  
Cell Counting ◽  
Migration And Invasion ◽  
Mtor Signaling Pathway ◽  
Invasion And Metastasis ◽  
Blotting Technique ◽  
Adhesion And Invasion

Background: Autophagy plays a "double-edged sword" in the process of tumorigenesis, development and metastasis. Objective: In this study, we explored the effect of PI3K/AKT/mTOR autophagy related signaling pathway on regulating and controlling the invasion and metastasis of liver cancer cells by Bufalin. Methods: The cell counting , migration , adhesion and invasion assay were used to evaluate the effect of Bufalin on the cell proliferation, invasion and metastasis. The protein expression of PI3K/AKT/mTOR signaling pathway were detected by Western Blotting technique. Results: After inhibiting autophagy of HCC-LM3 cells, the inhibitory effect of Bufalin on adhesion, migration and invasion of HCC-LM3 cells was significantly enhanced. The synergistic inhibition was strongest when different autophagy inhibitors were combined with 3MA and CQ. After inhibiting autophagy, Bufalin significantly inhibited the protein expression of P-AKT, Cyclin D1, MMP-2, MMP-9 and VEGF in HCC-LM3 cells. The protein expression of PTEN and E-Cadherin in HCC-LM3 cells was significantly increased. Conclusion: The present study shows that the anti-tumor effect of Bufalin mainly inhibit the proliferation, extracellular matrix degradation and angiogenesis of HCC by influencing autophagy. These findings confirm the capability of Bufalin in inhibiting metastasis of HCC and in parallel to current patents could be applied as a novel therapeutic strategy in the prevention of metastasis of HCC.

Sign in / Sign up

Export Citation Format

Share Document