scholarly journals H2S alleviates Parkinson-like phenotypes by modulating lncRNA-CasC7/miR-30c/BNIP3L signaling pathway

2021 ◽  
Author(s):  
Ling Long ◽  
Xiaodong Cai ◽  
Jinchi Liao ◽  
Xiaomeng Ma ◽  
Yingying Liu ◽  
...  
Keyword(s):  
Cell Culture ◽  
Signaling Pathway ◽  
Neuronal Apoptosis ◽  
Cell Model ◽  
Motor Deficits ◽  
Motor Disorder ◽  
Regulatory Relationship ◽  
Interacting Protein ◽  
Protein Levels ◽  
Culture Model

IntroductionThe pathogenesis of Parkinson’s disease (PD) is closely related to mitophagy, a process regulated by miRNAs and long non-coding RNAs (lncRNAs). In this study, we investigated the role of the lncRNA-CasC7/miR-30c/BNIP3L (BCL2 Interacting Protein 3 Like) signaling pathway in PD.Material and methodsA 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD was generated and treated by hydrogen sulfide (H2S) inhalation. The SH-SY5Y cell model of PD was generated by MPP+ and the treatment was NaHS. The open field, rotarod and tail suspension tests were performed to assess motor deficits. TUNEL and immunofluorescence assays were used to evaluate neuronal apoptosis in mice and in SH-SY5Y cell culture. Real-time PCR was performed to measure the expression level of the lncRNA-CasC7, miR-30c and BNIP3L, and western blotting was used to assess the protein levels of CBS, BNIP3L and PINK1. Luciferase assays were conducted to examine the regulatory relationship between miR-30c and lncRNA-CasC7/BNIP3L.ResultsH2S inhalation alleviated the motor disorder and neuronal apoptosis in PD mice, and NaHS treatment inhibited apoptosis in the SH-SY5Y cell culture model of PD. The sulfide compounds also ameliorated the dysregulated expression of CasC7, miR-30c, BNIP3L, and PINK1 in the PD models. Furthermore, miR-30c significantly inhibited the expression of lncRNA-CasC7 and BNIP3L, as assessed with the luciferase assays.ConclusionsOur findings suggest that the lncRNA-CasC7/miR-30c/BNIP3L mitophagy signaling pathway is involved in the pathogenesis of PD.

scholarly journals Mutant DMPK 3′-UTR transcripts disrupt C2C12 myogenic differentiation by compromising MyoD

2002 ◽  
Vol 159 (3) ◽  
pp. 419-429 ◽  
Author(s):  
Jeffrey D. Amack ◽  
Shannon R. Reagan ◽  
Mani S. Mahadevan
Keyword(s):  
Cell Culture ◽  
Myogenic Differentiation ◽  
Myoblast Differentiation ◽  
C2c12 Myoblast ◽  
Cell Culture Model ◽  
Rna Molecules ◽  
Protein Levels ◽  
Culture Model ◽  
Dystrophia Myotonica Protein Kinase ◽  
E Boxes

Myotonic dystrophy (DM) is caused by two similar noncoding repeat expansion mutations (DM1 and DM2). It is thought that both mutations produce pathogenic RNA molecules that accumulate in nuclear foci. The DM1 mutation is a CTG expansion in the 3′ untranslated region (3′-UTR) of dystrophia myotonica protein kinase (DMPK). In a cell culture model, mutant transcripts containing a (CUG)200 DMPK 3′-UTR disrupt C2C12 myoblast differentiation; a phenotype similar to what is observed in myoblast cultures derived from DM1 patient muscle. Here, we have used our cell culture model to investigate how the mutant 3′-UTR RNA disrupts differentiation. We show that MyoD protein levels are compromised in cells that express mutant DMPK 3′-UTR transcripts. MyoD, a transcription factor required for the differentiation of myoblasts during muscle regeneration, activates differentiation-specific genes by binding E-boxes. MyoD levels are significantly reduced in myoblasts expressing the mutant 3′-UTR RNA within the first 6 h under differentiation conditions. This reduction correlates with blunted E-box–mediated gene expression at time points that are critical for initiating differentiation. Importantly, restoring MyoD levels rescues the differentiation defect. We conclude that mutant DMPK 3′-UTR transcripts disrupt myoblast differentiation by reducing MyoD levels below a threshold required to activate the differentiation program.

scholarly journals Comparison of antioxidant activity between cyanidin-3-O-glucoside (C3G) liposome and cyanidin-3-O-glucoside (C3G) in 2D and 3D cell cultures

2018 ◽  
Author(s):  
Tisong Liang ◽  
Rongfa Guan ◽  
Guozhou Cao ◽  
Haitao Shen ◽  
Zhenfeng Liu ◽  
...  
Keyword(s):  
Cell Culture ◽  
Antioxidant Activity ◽  
Cell Cultures ◽  
Cell Model ◽  
3D Cell Culture ◽  
Culture Model ◽  
Mda Content ◽  
2D And 3D ◽  
Cells Cultured

ABSTRACTThe 2D cell culture is the predominant in vitro model for numerous studies. However, 2D cell cultures may not accurately reflect the functions of three-dimensional (3D) tissues, which have extensive cell–cell and cell–matrix interactions; thus, using 2D cell cultures may lead to inaccurate experimental results. Therefore, to obtain adequate and detailed information about the antioxidant activity of cyanidin-3-O-glucoside (C3G) and C3G liposomes in the 2D and 3D cell culture models, we used in this study H2O2to construct the cell damage model and assess the antioxidant activity of C3G and C3G liposomes on Caco-2 cells cultured in the 3D model. We also measured the cell viability, cell morphology, and activity of glutathione (GSH), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) content of Caco-2 cells treated with H2O2, C3G, and C3G liposomes. Results showed that cells cultured in the 3D culture model formed a 3D structure and tight spheroids and showed increased cell activity and IC50. The C3G and C3G liposomes can enhance the activity of GSH, SOD, and T-AOC but decrease the MDA content. At the same time, the effect was more obvious in the 3D cell culture model than in the cells cultured in the 2D model. This study revealed that the results obtained from the 2D cell model may be inaccurate compared with the results obtained from the 3D cell model. A realistic mechanism study of antioxidant activity of C3G and C3G liposomes in the 3D cell model, which acts as an intermediate stage bridging the in vitro 2D and in vivo models, was observed.

scholarly journals The Blood Component Iron Causes Neuronal Apoptosis Following Intracerebral Hemorrhage via the PERK Pathway

2020 ◽  
Vol 11 ◽  
Author(s):  
Muyao Wu ◽  
Rong Gao ◽  
Baoqi Dang ◽  
Gang Chen
Keyword(s):  
Intracerebral Hemorrhage ◽  
Signaling Pathway ◽  
Neuronal Apoptosis ◽  
Tunel Staining ◽  
Blood Component ◽  
Blood Components ◽  
Sprague Dawley ◽  
Neuroprotective Effects ◽  
Protein Levels ◽  
Neurological Score

PERK signaling pathway plays an important role in neuronal apoptosis after Intracerebral hemorrhage (ICH). ICH can cause the release of blood components into the brain. However, which component in the blood plays a major role still unclear. This study was designed to investigate the activation of the PERK pathway in different blood components after ICH and explore which components have major relationships with neuronal apoptosis. Eighty-five Sprague–Dawley rats were used to establish an ICH model. Western blot (WB) and immunofluorescence (IF) were used to evaluate the expression of the PERK pathway. TUNEL staining, FJC staining and neurological score were used to evaluate neuronal apoptosis and necrosis after ICH. The results showed that protein levels of p-PERK and p-eIF2α were upregulated following ICH with the injection of Fe3+ and Fe2+ after 48 h. Then, deferoxamine (DFX) was used to study the roles of Fe3+ in ICH through the PERK signaling pathway. The results showed that injection of DFX reversed increasing protein levels and prevented neuronal apoptosis. Thus, iron plays an important role in ICH through the PERK signaling pathway. Furthermore, the reduction of iron demonstrates neuroprotective effects in ICH. This suggests that targeting intervention of the iron and PERK pathway could be an effective treatment strategy to improve ICH prognosis.

scholarly journals Secreted Wnt6 mediates diabetes-associated centrosome amplification via its receptor FZD4

2020 ◽  
Vol 318 (1) ◽  
pp. C48-C62 ◽  
Author(s):  
Qin Ju He ◽  
Pu Wang ◽  
Qin Qin Liu ◽  
Qi Gui Wu ◽  
Yuan Fei Li ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Cell Culture ◽  
Signaling Pathway ◽  
Molecular Mechanisms ◽  
Nuclear Translocation ◽  
Integration Site ◽  
Cellular Protein ◽  
Centrosome Amplification ◽  
Protein Levels

We recently published that type 2 diabetes promotes cell centrosome amplification via upregulation of Rho-associated protein kinase 1 (ROCK1) and 14-3-3 protein-σ (14-3-3σ). This study further investigates the molecular mechanisms underlying diabetes-associated centrosome amplification. We found that treatment of cells with high glucose, insulin, and palmitic acid levels increased the intracellular and extracellular protein levels of Wingless-type MMTV integration site family member 6 (Wnt6) as well as the cellular level of β-catenin. The treatment also activated β-catenin and promoted its nuclear translocation. Treatment of cells with siRNA species for Wnt6, Frizzled-4 (FZD4), or β-catenin as well as introduction of antibodies against Wnt6 or FZD4 to the cell culture medium could all attenuate the treatment-triggered centrosome amplification. Moreover, we showed that secreted Wnt6-FZD4-β-catenin was the signaling pathway that was upstream of ROCK1 and 14-3-3σ. We found that advanced glycation end products (AGEs) were also able to increase the cellular and extracellular levels of Wnt6, the cellular protein level of β-catenin, and centrosome amplification. Treatment of the cells with siRNA species for Wnt6 or FZD4 as well as introduction of antibodies against Wnt6 or FZD4 to the cell culture could all inhibit the AGEs-elicited centrosome amplification. In colon tissues from a diabetic mouse model, the protein levels of Wnt6 and 14-3-3σ were increased. In conclusion, our results showed that the pathophysiological factors in type 2 diabetes, including AGEs, were able to induce centrosome amplification. It is suggested that secreted Wnt6 binds to FZD4 to activate the canonical Wnt6 signaling pathway, which is upstream of ROCK1 and 14-3-3σ, and that this is the cell signaling pathway underlying diabetes-associated centrosome amplification.

scholarly journals Bilobalide Suppresses Adipogenesis in 3T3-L1 Adipocytes via the AMPK Signaling Pathway

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3503 ◽  
Author(s):  
Su Bu ◽  
Chun Ying Yuan ◽  
Quan Xue ◽  
Ying Chen ◽  
Fuliang Cao
Keyword(s):  
Lipid Metabolism ◽  
Signaling Pathway ◽  
Ginkgo Biloba ◽  
Cell Model ◽  
Ginkgo Biloba Extract ◽  
Hormone Sensitive Lipase ◽  
Protective Effects ◽  
Ampk Signaling ◽  
Protein Levels ◽  
Compound C

Bilobalide, the only sesquiterpene compound from Ginkgo biloba leaf, exhibits various beneficial pharmaceutical activities, such as antioxidant, anti-inflammation, and protective effects for the central nervous system. Several bioactive components extracted from Ginkgo biloba extract reportedly have the potential to attenuate lipid metabolism. However, the effect of bilobalide on lipid metabolism remains unclear. In this study, we used 3T3-L1 cells as the cell model to investigate the effect of bilobalide on adipogenesis. The results showed that bilobalide inhibited 3T3-L1 preadipocyte differentiation and intracellular lipid accumulation. Quantitative real-time PCR and western blotting results indicated that several specific adipogenic transcription factors and a few important adipogenesis-related genes were significantly down regulated on both mRNA and protein levels in bilobalide treatment groups. By contrast, the expression of some lipolytic genes, such as adipose triglyceride lipase, hormone-sensitive lipase (HSL), and carnitine palmitoyltransferase-1α, were all up-regulated by bilobalide treatment, and the phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase 1, and HSL were stimulated. Furthermore, bilobalide treatment partially restored AMPK activity following its blockade by compound C (dorsomorphin). These results suggested that bilobalide inhibited adipogenesis and promoted lipolysis in 3T3-L1 cells by activating the AMPK signaling pathway.

1120: Citrate and Vitamin E Blunt the SWL-Induced Free Radical Surge in an In-Vitro MDCK Cell Culture Model

2004 ◽  
Vol 171 (4S) ◽  
pp. 295-295
Author(s):  
Fernando C. Delvecchio ◽  
Ricardo M. Brizuela ◽  
Karen J. Byer ◽  
W. Patrick Springhart ◽  
Saeed R. Khan ◽  
...  
Keyword(s):  
Cell Culture ◽  
Free Radical ◽  
Vitamin E ◽  
Mdck Cell ◽  
Cell Culture Model ◽  
Culture Model

Platelet-Rich and Platelet-Poor Plasma Might Play Supportive Roles in Cancer Cell Culture: A Replacement for Fetal Bovine Serum?

2021 ◽  
Vol 21 ◽  
Author(s):  
Mehdi Talebi ◽  
Mousa Vatanmakanian ◽  
Ali Mirzaei ◽  
Yaghoub Barfar ◽  
Maryam Hemmatzadeh ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Growth ◽  
Cancer Cell ◽  
Human Cancer ◽  
High Price ◽  
Dependent Manner ◽  
Platelet Poor Plasma ◽  
Protein Levels ◽  
Healthy Donors ◽  
Regulatory Functions

Background: Platelet-rich (PRP) and Platelet-poor plasma (PPP) are widely used in research and clinical platforms mainly due to their capacities to enhance cell growth. Although short half-life (5 days) and the high price of platelet products pose challenges regarding their usage, they maintain the growth regulatory functions for weeks. Thus, we aimed to assess the supplementary values of these products in human CCRF-CEM cancer cells. Mechanistically, we also checked if the PRP/PPP treatment enhances YKL-40 expression as a known protein regulating cell growth. Methods: The PRP/PPP was prepared from healthy donors using manual stepwise centrifugation and phase separation. The viability of the cells treated with gradient PRP/PPP concentrations (2, 5, 10, and 15%) was measured by the MTT assay. The YKL-40 mRNA and protein levels were assessed using qRT-PCR and western blotting. The data were compared to FBS-treated cells. Result: Our findings revealed that the cells treated by PRP/PPP not only were morphologically comparable to those treated by FBS but also, they showed greater viability at the concentrations of 10 and 15%. Moreover, it was shown that PRP/PPP induce cell culture support, at least in part, via inducing YKL-40 expression at both mRNA and protein levels in a time- and dose-dependent manner. Conclusion: Collectively, by showing cell culture support comparable to FBS, the PRP/PPP might be used as good candidates to supplement the cancer cell culture and overcome concerns regarding the use of FBS as a non-human source in human cancer research.

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