MiR-30b-5p Influences Chronic Exercise Arthritic Injury by Targeting Hoxa1

2021 ◽  
Author(s):  
Maoxun Li ◽  
Fei Gai ◽  
Hongyu Chen
Keyword(s):  
Caspase 3 ◽  
Cartilage Lesion ◽  
Type Ii Collagen ◽  
Cartilage Degradation ◽  
Chondrocyte Apoptosis ◽  
Chronic Exercise ◽  
Tnf Α ◽  
And Migration ◽  
Induced Apoptosis ◽  
Safranin O

AbstractWe identified the role of miR-30b-5p in chronic exercise arthritic injury. Rats with chronic exercise arthritic injury received treatment with miR-30b-5p antagomiR. H&E and Safranin O-fast green staining were performed. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected. The binding relationship between homeobox A1 (Hoxa1) and miR-30b-5p was revealed. After manipulating the expressions of miR-30b-5p and/or Hoxa1 in chondrocytes, the viability, apoptosis and migration of chondrocytes were assessed. The levels of molecules were determined by qRT-PCR or Western blot. MiR-30b-5p antagomiR ameliorated articular cartilage lesion and destruction, reduced Mankin’s score and the levels of TNF-α, IL-1β, miR-30b-5p, matrix metallopeptidase 13 (MMP-13), and cleaved caspase-3, and increased relative thickness and the levels of Hoxa1, Aggrecan and type II collagen (COLII) in model rats. MiR-30b-5p up-regulation decreased Hoxa1 level, viability, migration and induced apoptosis, whereas miR-30b-5p down-regulation produced the opposite effects. MiR-30b-5p up-regulation increased the levels of MMP-13 and cleaved caspase-3, but decreased those of Aggrecan and COLII in chondrocytes. However, the action of miR-30b-5p up-regulation on chondrocytes was reversed by Hoxa1 overexpression. In conclusion, miR-30b-5p is involved in cartilage degradation in rats with chronic exercise arthritic injury and regulates chondrocyte apoptosis and migration by targeting Hoxa1.

scholarly journals miR-27b-3p Targeting BDNF Inhibits the TrkB/CREB Signaling Pathway and Improves IL-1 β-Induced Chondrocyte Inflammation

2021 ◽  
Author(s):  
cailian ruan ◽  
Rui jun CONG ◽  
Miao Wang ◽  
LI JUN WANG ◽  
YONG YU ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Signaling Pathway ◽  
Cartilage Degradation ◽  
Camp Response Element Binding ◽  
Detection Methods ◽  
Chondrocyte Apoptosis ◽  
Cell Transfection ◽  
Expression Levels ◽  
Tnf Α ◽  
Safranin O

Abstract Background: Explore the mechanism of "miR-27b-3p targeting BDNF inhibits the TrkB/CREB signaling pathway and improves IL-1 β-induced chondrocyte inflammation".Methods: The animal and cell models of arthritis were constructed, and various biochemical detection methods were used to detect the changes of apoptosis, inflammation and oxidative stress.Results: The results showed that the expression of miR-27b-3p was downregulated in IL-1β-treated chondrocytes and cartilage tissues isolated from a KOA rat model. miR-27b-3p overexpression notably reduced IL-1β-induced chondrocyte apoptosis and the expression levels of caspase-3 and caspase-9. In addition, cell transfection with miR-27b-3p mimics increased the mRNA and protein expression levels of inducible nitric oxide synthase and cyclooxygenase-2. The levels of nitric oxide, prostaglandin E2 (PGE2), TNF-α and IL-6 were also reduced following cell transfection with miR-27b-3p mimics. Furthermore, bioinformatics analysis predicted that miR-27b-3p could directly target brain-derived neurotrophic factor (BDNF). Additionally, the present study suggested that the tropomyosin receptor kinase B (TrkB)/cAMP response‑element binding protein (CREB) signaling axis could be a downstream pathway of the miR-27b-3p/BDNF axis. The percentage of apoptotic cells and the expression levels of nitric oxide, PGE2, TNF-α and IL-6 were enhanced in chondrocytes co-treated with BDNF + IL-1β. However, these effects were restored following transfection of chondrocytes with miR-27b-3p mimics. Staining of cartilage tissues with safranin O showed that miR-27b-3p overexpression Significantly attenuated KOA-induced cartilage degradation.Conclusions:miR-27b-3p targeting BDNF inhibits the TrkB/CREB signaling pathway and improves IL-1 β-induced chondrocyte inflammation.

scholarly journals cIAP1 promotes proliferation and migration and prevents apoptosis in gallbladder cancer in vitro

2019 ◽  
Vol 39 (4) ◽  
Author(s):  
Wei Su ◽  
Xiaojie Jiang ◽  
Mingyuan Chen ◽  
Maotuan Huang ◽  
Nanhong Tang ◽  
...  
Keyword(s):  
Gallbladder Cancer ◽  
Protein C ◽  
Caspase 3 ◽  
Nuclear Factor Κb ◽  
Inhibitory Protein ◽  
Tnf Α ◽  
And Migration ◽  
Induced Apoptosis ◽  
Proliferation And Migration

Abstract Gallbladder cancer (GBC) is a demanding fatal disease with no ideal treatment for inoperable patients. Recent reports have determined TNF-α associated lymphatic metastasis in GBC, while its resistance to TNF-α-killing remains largely unexplored. In this assay, we first found cellular inhibitor of apoptosis (cIAP1) overexpressed in GBC tissues and the roles in promoting the proliferation and migration of GBC in vitro as its homology cIAP2 does. Then how GBC cell survives TNF-α toxicity and TNF-α-induced apoptosis first prevail as follows. The reduction in cIAP1 does not give rise to apoptosis even with the stimulation of TNF-α. Importantly, the loss of cIAP1 enhanced TNF-α/cycloheximide-induced apoptosis in higher activation statuses of Caspase-8, Caspase-3 without the induction of Complex Ⅱ. In response to TNF-α, the reduction in cIAP1 caused the suppression in nuclear factor-κB (NF-κB) pathway and inhibition of transcription of cell death regulator cellular FLICE-like Inhibitory Protein (c-FLIP) instead. To conclude, cIAP1 is an oncological protein abundant in GBC tissues, which enhances proliferation and immigration and blocks TNF-α from apoptosis through NF-κB pathway in vitro.

Paeonol Ameliorates Inflammation and Cartilage of Chondrocytes in Osteoarthritis by Upregulating SIRT1

2020 ◽  
Author(s):  
Peng Shang ◽  
Ying Liu ◽  
Junqing Jia
Keyword(s):  
Extracellular Matrix ◽  
Caspase 3 ◽  
Type Ii Collagen ◽  
Blue Staining ◽  
Chondrocyte Apoptosis ◽  
Matrix Degradation ◽  
Extracellular Matrix Degradation ◽  
Tnf Α ◽  
And Cartilage

Abstract Objective: To explore the possible role of paeonol on chondrocyte inflammation and cartilage protection in osteoarthritis (OA).Methods: Primary chondrocytes were isolated from rat stifle joints, and were identified through toluidine blue staining and immunofluorescence staining of type II collagen. The chondrocytes were transfected with sh-SIRT1 or/and paeonol (0, 20, 50, 100, 200, 1000 mg/L) before OA modeling induced by IL-1β. ELISA determined the expressions of TNF-α, IL-17 and IL-6, and apoptotic rate was examined by flow cytometry. qRT-PCR and Western blot quantified the expressions of MMP-1, MMP-3, MMP-13, TIMP-1, cleaved-caspase-3, Bax, Bcl-2, and the proteins related to NF-κB pathway.Results: Increases in TNF-α, IL-17, IL-6, MMP-1, MMP-3 and MMP-13 and decrease in TIMP-1 were found in IL-1β stimulated chondrocytes. The apoptotic rate as well as the expressions of cleaved-caspase-3 and Bax was up-regulated, and Bcl-2 expression was suppressed in response to IL-1β treatment. NF-κB pathway was activated in IL-1β-stimulated chondrocytes. Paeonol enhanced SIRT1 expression to inactivate NF-κB pathway, thus ameliorating the secretion of inflammatory cytokines, extracellular matrix degradation and chondrocyte apoptosis.Conclusion: Paeonol inhibits IL-1β induced inflammation and extracellular matrix degradation in chondrocytes through up-regulating SIRT1 and suppressing NF-κB pathway.

scholarly journals Apoptosis of Growth Plate Chondrocytes Occurs through a Mitochondrial Pathway

2007 ◽  
Vol 77 (1) ◽  
pp. 129-134 ◽  
Author(s):  
Cristina C. Teixeira ◽  
Aida P. Padron Costas ◽  
Yelena Nemelivsky
Keyword(s):  
Cytochrome C ◽  
Growth Plate ◽  
Caspase 3 ◽  
Mitochondrial Pathway ◽  
Chondrocyte Apoptosis ◽  
Cytochrome C Release ◽  
Growth Plate Chondrocytes ◽  
Chondrocyte Viability ◽  
Induced Apoptosis ◽  
Free Media

Abstract Objective: To determine the role of mitochondria in chondrocyte apoptosis induced by inorganic phosphate (Pi). Materials and Methods: Chondrocytes isolated from the growth plates of chick embryo tibia were treated with Pi in serum-free media; chondrocyte viability, mitochondrial membrane potential, cytochrome c release from mitochondria, caspase 3 activity, endonuclease activity, and DNA fragmentation were investigated. Results: Exposure to Pi for 24 hours induced apoptosis in growth plate chondrocytes through a pathway that involved loss of mitochondrial function, release of cytochrome c into the cytoplasm, increases in caspase 3 and endonuclease activities, and fragmentation of DNA. Conclusions: This study suggests that mitochondria are important players in Pi-induced apoptosis.

scholarly journals Blocking Caspase-3-Mediated Proteolysis of IKKβ Suppresses TNF-α-Induced Apoptosis

2001 ◽  
Vol 8 (5) ◽  
pp. 1005-1016 ◽  
Author(s):  
Guilin Tang ◽  
Jun Yang ◽  
Yuzuru Minemoto ◽  
Anning Lin
Keyword(s):  
Caspase 3 ◽  
Tnf Α ◽  
Induced Apoptosis

scholarly journals MiR-27b-3p Targeting BDNF Inhibits TrkB / CREB Signaling Pathway and Improves IL-1 β Induced Chondrocytic Inflammation

2021 ◽  
Author(s):  
Cailian Ruan ◽  
Rui jun Cong ◽  
Miao Wang ◽  
Lijun Wang ◽  
Yong Yu ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Apoptotic Cell ◽  
Cartilage Degradation ◽  
Chondrocyte Apoptosis ◽  
Caspase 9 ◽  
Treatment Target ◽  
Cell Percentage ◽  
Safranin O ◽  
Creb Pathway

Knee Osteoarthritis (KOA) is a chronic disease characterized by progressive disability and joint pain. Meniscus chondrocytes apoptosis is the main cause of reduced chondrocyte number and self-repair function. The purpose of this study was to investigate the role of miR-27b-3p in KOA.In this study, we found that the expression of miR-27b-3p was downregulated in cultured IL-1β treated chondrocyte and cartilage tissues in KOA. KOA overexpression evidently reduced IL-1β induced chondrocyte apoptosis and caspase-3 and caspase-9 expression.The upregulated iNOS and COX-2 mRNA and proteins expression was also inhibited by miR-27b-3p mimics. The expression of nitric oxide, PGE2, TNF-α and IL-6 was also inhibited by miR-27b-3p mimics. The target gene of miR-27b-3p was confirmed to be BDNF. TrkB/CREB pathway was proved to be the downstream pathway of miR-27b-3p/BDNF axis.The apoptotic cell percentage and nitric oxide, PGE2, TNF-α and IL-6 expression was induced by BDNF+IL-1β. This induction was inhibited by miR-27b-3p mimics. The cartilage tissues stained with safranin O results showed miR-27b-3p greatly decreased KOA induced cartilage degradation. The expression of BDNF、TrkB and p-CREB was inhibited by len-miR-27b-3p. MiR-27b-3p also reduced the expression of TNF-α、IL-6 and Bax, and increased Bcl-2 expression. These results indicated miR-27b-3p could applied to inhibit the development of KOA and miR-27b-3p/BDNF/TrkB/CREB pathway could serve as novel treatment target to handle KOA.

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 hnRNP K plays a protective role in TNF-α-induced apoptosis in podocytes

2018 ◽  
Vol 38 (3) ◽  
Author(s):  
Shili Zhao ◽  
Junxia Feng ◽  
Qi Wang ◽  
Lu Tian ◽  
Yunfang Zhang ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Glycogen Synthase ◽  
Kidney Diseases ◽  
Cell Cycle Distribution ◽  
Caspase 8 ◽  
G1 Arrest ◽  
Inhibitory Protein ◽  
Hnrnp K ◽  
Tnf Α ◽  
Induced Apoptosis

Apoptosis of podocytes contributes to proteinuria in many chronic kidney diseases. The cytokine, tumor necrosis factor-α (TNF-α) is thought to be involved in podocyte apoptosis, but the underlying mechanism is not understood. In our study, we established a model of TNF-α-induced apoptosis by isolating primary podocytes from mice. After exposing cells to TNF-α, we determined the expression levels of heterogeneous nuclear ribonucleoprotein K (hnRNP K) and cellular FLICE-inhibitory protein (c-FLIP) and the phosphorylation levels of glycogen synthase kinase β (GSK3β) and extracellular signal-regulated kinase (ERK). We then knocked down or overexpressed the levels of hnRNP K and observed its effects on the expressions of c-FLIP, caspase-8, caspase-3, and the phosphorylation of GSK3β and ERK. In addition, we examined the percentage of cells undergoing apoptosis and studied cell cycle distribution. We found that TNF-α induced apoptosis in podocytes and that the expressions of hnRNP K and c-FLIP were significantly decreased, whereas the phosphorylations of GSK3β and ERK were significantly increased. Both gene knockdown and overexpression of hnRPN K resulted in varied expressions/phosphorylations of c-FLIP, GSK3β, and ERK. Moreover, decreased hnRPN K expression contributed to increased levels of caspase-8 and capase-3, as well as an increase in cell apoptosis and G0/G1 arrest. In conclusion, down-regulated expression of hnRNP K by TNF-α resulted in a decrease in the expression of c-FLIP as well as increases in phosphorylated GSK3β, ERK, caspase-8, and caspase-3, and then critically contributed to the podocyte apoptosis.

NF-κB-mediated IAP expression induces resistance of intestinal epithelial cells to apoptosis after polyamine depletion

2004 ◽  
Vol 286 (5) ◽  
pp. C1009-C1018 ◽  
Author(s):  
Tongtong Zou ◽  
Jaladanki N. Rao ◽  
Xin Guo ◽  
Lan Liu ◽  
Huifang M. Zhang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Caspase 3 ◽  
Intestinal Epithelial Cells ◽  
Specific Inhibitor ◽  
Inhibitory Effect ◽  
Intestinal Epithelial ◽  
Tnf Α ◽  
Induced Apoptosis ◽  
Necrosis Factor ◽  
Active Caspase

Apoptosis plays a crucial role in maintenance of intestinal epithelial integrity and is highly regulated by numerous factors, including cellular polyamines. We recently showed that polyamines regulate nuclear factor (NF)-κB activity in normal intestinal epithelial (IEC-6) cells and that polyamine depletion activates NF-κB and promotes resistance to apoptosis. The current study went further to determine whether the inhibitors of apoptosis (IAP) family of proteins, c-IAP2 and XIAP, are downstream targets of activated NF-κB and play a role in antiapoptotic activity of polyamine depletion in IEC-6 cells. Depletion of cellular polyamines by α-difluoromethylornithine not only activated NF-κB activity but also increased expression of c-IAP2 and XIAP. Specific inhibition of NF-κB by the recombinant adenoviral vector containing IκBα superrepressor (Ad Iκ BSR) prevented the induction of c-IAP2 and XIAP in polyamine-deficient cells. Decreased levels of c-IAP2 and XIAP proteins by inactivation of NF-κB through Ad Iκ BSR infection or treatment with the specific inhibitor Smac also overcame the resistance of polyamine-depleted cells to apoptosis induced by the combination of tumor necrosis factor (TNF)-α and cycloheximide (CHX). Although polyamine depletion did not alter levels of procaspase-3 protein, it inhibited formation of the active caspase-3. Decreased levels of c-IAP2 and XIAP by Smac prevented the inhibitory effect of polyamine depletion on the cleavage of procaspase-3 to the active caspase-3. These results indicate that polyamine depletion increases expression of c-IAP2 and XIAP by activating NF-κB in intestinal epithelial cells. Increased c-IAP2 and XIAP after polyamine depletion induce the resistance to TNF-α/CHX-induced apoptosis, at least partially, through inhibition of the caspase-3 activity.

scholarly journals Cleavage of Cdc6 by caspase-3 promotes ATM/ATR kinase–mediated apoptosis of HeLa cells

2006 ◽  
Vol 174 (1) ◽  
pp. 77-88 ◽  
Author(s):  
Hyungshin Yim ◽  
In Sun Hwang ◽  
Joon-Seok Choi ◽  
Kwang-Hoon Chun ◽  
Ying Hua Jin ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Replication Initiation ◽  
Dominant Negative ◽  
Ataxia Telangiectasia Mutated ◽  
Activation Kinetics ◽  
Kinase Activation ◽  
Tnf Α ◽  
Induced Apoptosis ◽  
Atr Kinase ◽  
Interfering Rna

We show that caspase-3 cleaves Cdc6 at D290/S and D442/G sites, producing p32-tCdc6 (truncated Cdc6) and p49-tCdc6, respectively, during etoposide- or tumor necrosis factor (TNF)-α–induced apoptosis. The expression of these tCdc6 proteins, p32- and p49-tCdc6, promotes etoposide-induced apoptosis. The expression of tCdc6 perturbs the loading of Mcm2 but not Orc2 onto chromatin and activates ataxia telangiectasia mutated (ATM) and ATM and Rad-3 related (ATR) kinase activities with kinetics similar to that of the phosphorylation of Chk1/2. The activation kinetics are consistent with elevated cellular levels of p53 and mitochondrial levels of Bax. The tCdc6-induced effects are all suppressed to control levels by expressing a Cdc6 mutant that cannot be cleaved by caspase-3 (Cdc6-UM). Cdc6-UM expression attenuates the TNF-α–induced activation of ATM and caspase-3 activities. When ATM or ATR is down-expressed by using the small interfering RNA technique, the TNF-α– or tCdc6-induced activation of caspase-3 activities is suppressed in the cells. These results suggest that tCdc6 proteins act as dominant-negative inhibitors of replication initiation and that they disrupt chromatin structure and/or induce DNA damage, leading to the activation of ATM/ATR kinase activation and p53–Bax-mediated apoptosis.

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