Inhibition of long noncoding RNA MALAT1 suppresses high glucose-induced apoptosis and inflammation in human umbilical vein endothelial cells by suppressing the NF-κB signaling pathway

2020 ◽  
Vol 98 (6) ◽  
pp. 669-675
Author(s):  
Yu-Ping Gong ◽  
Ya-Wei Zhang ◽  
Xiao-Qing Su ◽  
Hai-Bo Gao
Keyword(s):  
Endothelial Cells ◽  
High Glucose ◽  
Long Noncoding Rna ◽  
Noncoding Rna ◽  
Caspase 3 ◽  
Caspase 9 ◽  
Expression Levels ◽  
Tnf Α ◽  
Induced Apoptosis ◽  
Apoptosis And Inflammation

The study investigated the expression of long noncoding RNA (lncRNA) MALAT1 in high glucose (HG)-induced human vascular endothelial cells (HUVECs) and the role of MALAT1 in the apoptosis of HG-induced HUVECs. The HUVECs were cultured and induced with 25 mmol/L HG. After that, the HUVECs were transfected with MALAT1 siRNA. The expression levels of MALAT1 were detected with qPCR, whereas the expression levels of Bax, Bcl-2, cleaved-caspase-3, cleaved-caspase-9, p-65, and p-p65 were detected using Western blot. The roles of MALAT1 in cell activities, including apoptosis, were evaluated using the CCK-8 assay, TUNEL staining, and flow cytometry. The expression levels of inflammatory factors (TNF-α and IL-6) were measured using ELISA. The expression levels of MALAT1, TNF-α, and IL-6 in HUVECs were increased in the HG environment; however, when MALAT1 was silenced in the HUVECs, cell proliferation increased significantly, the expression levels of TNF-α, IL-6, Bax, cleaved-caspase-3, and cleaved-caspase-9 decreased, and the rate of apoptosis also decreased. Silencing MALAT1 inhibited the expression of p-p65 in HG-induced HUVECs. In conclusion, our study demonstrated that MALAT1 is upregulated in HG-induced HUVECs, and inhibition of MALAT1 inhibits HG-induced apoptosis and inflammation in HUVECs by suppression of the NF-κB signaling pathway.

scholarly journals Pearl extract protects HaCaT cells from UV radiation-induced apoptosis through mitochondrial pathway regulation

2020 ◽  
Author(s):  
Zhixiong Chen ◽  
jing wang ◽  
Anquan Yang ◽  
Lihua Zhang ◽  
Yaojia Lu ◽  
...  
Keyword(s):  
Protein Expression ◽  
Caspase 3 ◽  
Hacat Cells ◽  
Caspase 9 ◽  
Expression Levels ◽  
Protein Levels ◽  
Bax Protein ◽  
Reactive Oxygen Species Content ◽  
Tnf Α ◽  
Induced Apoptosis

Abstract Background: Previous studies have demonstrated that pearl extract (PE) promotes wound healing and skin whitening. However, it remains unclear whether PE can inhibit ultraviolet (UV)-photodamage in HaCaT cells. In this study, an in vitro photoaging cell model was established to observe the effect of PE on UV-induced damage and the apoptosis of HaCaT cells. The aim of this study was to provide a reference for the future development of natural sunscreens.Results: PE concentrations of 0.1 and 1 μg/mL were considered the most effective and safe concentrations. Compared to that in the control group, superoxide dismutase and glutathione peroxidase activity in the photoaging group was significantly reduced, whereas malondialdehyde and reactive oxygen species content, along with tumour necrosis factor-alpha (TNF-α) and interleukin (IL)-10 mRNA and protein levels, were markedly increased. In contrast, Bcl-2 protein expression was significantly decreased, whereas caspase-3, caspase-9, and Bax protein expression levels were significantly increased. Compared to that in the photoaging group, HaCaT cell proliferation was significantly increased in the PE group. Both PE concentrations significantly increased superoxide dismutase and glutathione peroxidase activity in cells, reduced malondialdehyde and reactive oxygen species content, decreased TNF-α and IL-10 mRNA expression in cells, and reduced TNF-α and IL-10 protein levels in the supernatant. Additionally, Bcl-2 protein expression levels were significantly increased, whereas caspase-3, caspase-9, and Bax protein expression levels were significantly reduced by PE treatment.Conclusions: PE can inhibit UV-induced apoptosis by inhibiting mitochondria-mediated apoptosis and regulating TNF-α and IL-10 expression.

Polyamines are required for activation of c-Jun NH2-terminal kinase and apoptosis in response to TNF-α in IEC-6 cells

2003 ◽  
Vol 285 (5) ◽  
pp. G980-G991 ◽  
Author(s):  
Sujoy Bhattacharya ◽  
Ramesh M. Ray ◽  
Mary Jane Viar ◽  
Leonard R. Johnson
Keyword(s):  
Cytochrome C ◽  
Dna Fragmentation ◽  
Caspase 3 ◽  
Specific Inhibitor ◽  
Cell Types ◽  
Caspase 9 ◽  
Cytochrome C Release ◽  
Tnf Α ◽  
Induced Apoptosis ◽  
Jnk Activation

Intracellular polyamine homeostasis is important for the regulation of cell proliferation and apoptosis and is necessary for the balanced growth of cells and tissues. Polyamines have been shown to play a role in the regulation of apoptosis in many cell types, including IEC-6 cells, but the mechanism is not clear. In this study, we analyzed the mechanism by which polyamines regulate the process of apoptosis in response to tumor necrosis factor-α (TNF-α). TNF-α or cycloheximide (CHX) alone did not induce apoptosis in IEC-6 cells. Significant apoptosis was observed when CHX was given along with TNF-α, as indicated by a significant increase in the detachment of cells, caspase-3 activity, and DNA fragmentation. Polyamine depletion by treatment with α-difluoromethylornithine significantly reduced the level of apoptosis, as judged by DNA fragmentation and the caspase-3 activity of attached cells. Apoptosis in IEC-6 cells was accompanied by the activation of upstream caspases-6, -8, and -9 and NH2-terminal c-Jun kinase (JNK). Inhibition of JNK activation prevented caspase-9 activation. Polyamine depletion prevented the activation of JNK and of caspases-6, -8, -9, and -3. SP-600125, a specific inhibitor of JNK activation, prevented cytochrome c release from mitochondria, JNK activation, DNA fragmentation, and caspase-9 activation in response to TNF-α/CHX. In conclusion, we have shown that polyamine depletion delays and decreases TNF-α-induced apoptosis in IEC-6 cells and that apoptosis is accompanied by the release of cytochrome c, the activation of JNK, and of upstream caspases as well as caspase-3. Polyamine depletion prevented JNK activation, which may confer protection against apoptosis by modulation of upstream caspase-9 activation.

scholarly journals Pearl extract protects HaCaT cells from UV radiation-induced apoptosis through mitochondrial pathway regulation

2019 ◽  
Author(s):  
Zhixiong Chen ◽  
Jing Wang ◽  
Anquan Yang ◽  
Lihua Zhang ◽  
Yaojia Lu ◽  
...  
Keyword(s):  
Protein Expression ◽  
Caspase 3 ◽  
Hacat Cells ◽  
Oxygen Species ◽  
Caspase 9 ◽  
Expression Levels ◽  
Protein Levels ◽  
Bax Protein ◽  
Reactive Oxygen Species Content ◽  
Induced Apoptosis

Abstract Background: Previous studies demonstrated that pearl extract (PE) promotes wound healing and skin whitening. However, whether PE can inhibit ultraviolet (UV) photodamage in HaCaT cells remains unclear. In this study, an in vitro photoaging cell model was established to observe the effect of PE on UV-induced damage and apoptosis of HaCaT cells. The aim was to provide a reference for future development of natural sunscreen agents. Results: PE concentrations of 0.1 and 1 μg/mL were considered as the most effective and safe concentrations. Compared to the control group, superoxide dismutase and glutathione peroxidase activities in the photoaging group were significantly reduced, while malondialdehyde and reactive oxygen species content, along with tumor necrosis factor-alpha (TNF-a) and interleukin (IL)-10 mRNA and protein levels were markedly increased. In contrast, Bcl-2 protein expression was significantly decreased, while caspase-3, caspase-9, and Bax protein expression levels were significantly increased. Compared to the photoaging group, HaCaT cell proliferation was significantly increased in the PE group. Both PE concentrations significantly increased superoxide dismutase and glutathione peroxidase activities in cells, reduced malondialdehyde and reactive oxygen species content, decreased TNF-a and IL-10 mRNA expression in cells, and reduced TNF-a and IL-10 protein levels in the supernatant. Additionally, Bcl-2 protein expression levels were significantly increased, while caspase-3, caspase-9, and Bax protein expression levels were significantly reduced by PE treatment. Conclusions: PE can inhibit UV-induced apoptosis by inhibiting mitochondria-mediated apoptosis and regulating TNF-a and IL-10 expression.

scholarly journals High Glucose–Induced Apoptosis in Human Endothelial Cells Is Mediated by Sequential Activations of c-Jun NH 2 -Terminal Kinase and Caspase-3

Circulation ◽  
2000 ◽  
Vol 101 (22) ◽  
pp. 2618-2624 ◽  
Author(s):  
Feng M. Ho ◽  
Shing H. Liu ◽  
Chiau S. Liau ◽  
Por J. Huang ◽  
Shoei Y. Lin-Shiau
Keyword(s):  
Endothelial Cells ◽  
High Glucose ◽  
Caspase 3 ◽  
Human Endothelial Cells ◽  
Induced Apoptosis

scholarly journals Long noncoding RNA KCNQ1OT1 induces pyroptosis in diabetic corneal endothelial keratopathy

2020 ◽  
Vol 318 (2) ◽  
pp. C346-C359 ◽  
Author(s):  
Yanyan Zhang ◽  
Zhen Song ◽  
Xuran Li ◽  
Shuo Xu ◽  
Sujun Zhou ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
High Glucose ◽  
Long Noncoding Rna ◽  
Noncoding Rna ◽  
Rna Binding ◽  
Cultured Cells ◽  
Corneal Edema ◽  
Corneal Endothelial Cells ◽  
Caspase 1

Diabetic corneal endothelial keratopathy is an intractable ocular complication characterized by corneal edema and endothelial decompensation, which seriously threaten vision. It has been suggested that diabetes is associated with pyroptosis, a type of programmed cell death via the activation of inflammation. Long noncoding RNA KCNQ1OT1 is commonly associated with various pathophysiological mechanisms of diabetic complications, including diabetic cardiomyopathy and diabetic retinopathy. However, whether KCNQ1OT1 is capable of regulating pyroptosis and participates in the pathogenesis of diabetic corneal endothelial keratopathy remains unknown. The aim of this study was to investigate the mechanisms of KCNQ1OT1 in diabetic corneal endothelial keratopathy. Here, we reveal that KCNQ1OT1 and pyroptosis can be triggered in diabetic human and rat corneal endothelium, along with the high glucose-treated corneal endothelial cells. However, miR-214 expression was substantially decreased in vivo and in experiments with cultured cells. LDH assay was also used to verify the existence of pyroptosis in high glucose-treated cells. Bioinformatics prediction and luciferase assays showed that KCNQ1OT1 may function as a competing endogenous RNA binding miR-214 to regulate the expression of caspase-1. To further analyze the KCNQ1OT1-mediated mechanism, miR-214 mimic and inhibitor were introduced into the high glucose-treated corneal endothelial cells. The results showed that upregulation of miR-214 attenuated pyroptosis; conversely, knockdown of miR-214 promoted it. In addition, KCNQ1OT1 knockdown by a small interfering RNA decreased pyroptosis factors expressions but enhanced miR-214 expression in corneal endothelial cells. To understand the signaling mechanisms underlying the prepyroptotic properties of KCNQ1OT1, si-KCNQ1OT1 was cotransfected with or without miR-214 inhibitor. The results showed that pyroptosis was repressed after silencing KCNQ1OT1 but was reversed by cotransfection with miR-214 inhibitor, suggesting that KCNQ1OT1 mediated pyroptosis induced by high glucose via targeting miR-214. Therefore, the KCNQ1OT1/miR-214/caspase-1 signaling pathway represents a new mechanism of diabetic corneal endothelial keratopathy progression, and KCNQ1OT1 could potentially be a novel therapeutic target.

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