scholarly journals miR-223 Inhibits the Polarization and Recruitment of Macrophages via NLRP3/IL-1β Pathway to Meliorate Neuropathic Pain

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Junsong Zhu ◽  
Jinmei Yang ◽  
Jianguo Xu
Keyword(s):  
Neuropathic Pain ◽  
Underlying Mechanism ◽  
Inflammatory Factor ◽  
Mice Model ◽  
Protein Levels ◽  
Caspase 1 ◽  
Luciferase Activity Assay ◽  
Macrophage Chemotaxis ◽  
Group Flow

Background. miRNA is an essential factor in neuropathic pain. However, the underlying mechanism of miRNA in neuropathic pain remains unclear. Objective. To explore the potential role of miR-223 in neuropathic pain in a mice model of chronic sciatic nerve injury. Methods. Mice were divided into the sham group, CCI group, CCI + Lenti-vector group, and CCI + Lenti-miR-223 group. Flow cytometry was used to detect the neuronal apoptosis and the proportion of M1/M2 macrophages in each group. Western blot was used to detect the protein expression levels of ASC, caspase-1, IL-1β, and IL-18 in each group. Luciferase activity assay detects the binding of miR-223 and NLRP3. Macrophage chemotaxis experiments verified the anti-inflammatory effect of miR-223 in vitro. Results. The overexpression of miR-233 significantly reduced the neuropathic pain caused by CCI and reduced the apoptosis and inflammatory factor expression. miR-223 inhibits the expression of NLRP3 by directly binding to the 3′-untranslated region. Overexpression of miR-223 reduces the protein levels of NLRP3, ASC, caspase-1, IL-1β, and IL-18 in the spinal cord of CCI mice, increases the proportion of M2-type macrophages, and reduces the proportion of M1-type macrophages. Conclusion. miR-223 may facilitate the development of neuropathic pain in CCI mice by inhibiting NLRP3-mediated neuroinflammation.

scholarly journals Callicarpa Nudiflora Extract Exerts Anti-Inflammatory Effect on H. Pylori-Associated Gastritis Through Repression of ROS/NLRP3/Caspase-1/IL-1β Signaling Axis

2021 ◽  
Author(s):  
Lili Li ◽  
Xiaohui Zhu ◽  
Xingxing Chai ◽  
Xiaoyu Chen ◽  
Xiaohua Su ◽  
...  
Keyword(s):  
Cell Damage ◽  
Underlying Mechanism ◽  
Inflammatory Factors ◽  
Inflammasome Activation ◽  
Ros Production ◽  
Gastric Diseases ◽  
Caspase 1 ◽  
Signaling Axis ◽  
H Pylori

Abstract Helicobacter pylori ( H. pylori ) is a major pathogenic factor for the development of gastric diseases including chronic gastritis and gastric cancer. Callicarpa nudiflora (CN), an air-dried leaves extract of Callicarpa nudiflora Hook. & Arn., has been found to exhibit a broad-spectrum antibacterial effect. In our study, we extracted the active ingredient from air-dried leaves of Callicarpa nudiflora, detected the effect of CN against H. pylori -infected GES-1 cells in vitro , and elucidated the underlying mechanism. GES-1 cells were cocultured with HPSS1 at MOI = 100:1 and treated with different concentrations of CN. Results indicated that CN not only significantly decreased cellular lactate dehydrogenase leakage, but also markedly attenuated H. pylori -induced cell apoptosis and ROS production in GSE-1 cells, therefore protecting gastric epithelial cells against injuries caused by H. pylori . CN also inhibited the secretions of inflammatory factors, such as tumor necrosis factor-α (TNF-α), IL-1β, IL-6 and IL-8. Furthermore, CN remarkably decreased the expression levels of NLRP3, PYCARD, active Caspase-1. In conclusion, CN exhibited highly efficient protective effect against H. pylori -induced gastritis and cell damage; Mechanismly, CN suppressed H. pylori -triggered inflammatory response and pyroptosis through depressing ROS production and NLRP3 inflammasome activation via ROS/NLRP3/IL-1β signaling axis.

scholarly journals hnRNP K Supports High-Amplitude D Site-Binding Protein mRNA (Dbp mRNA) Oscillation To Sustain Circadian Rhythms

2020 ◽  
Vol 40 (6) ◽  
Author(s):  
Paul Kwangho Kwon ◽  
Kyung-Ha Lee ◽  
Ji-hyung Kim ◽  
Sookil Tae ◽  
Seokjin Ham ◽  
...  
Keyword(s):  
Clock Genes ◽  
Binding Protein ◽  
Underlying Mechanism ◽  
High Amplitude ◽  
Specific Protein ◽  
Proximal Promoter ◽  
Hnrnp K ◽  
Protein Levels ◽  
Site Binding

ABSTRACT Circadian gene expression is defined by the gene-specific phase and amplitude of daily oscillations in mRNA and protein levels. D site-binding protein mRNA (Dbp mRNA) shows high-amplitude oscillation; however, the underlying mechanism remains elusive. Here, we demonstrate that heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a key regulator that activates Dbp transcription via the poly(C) motif within its proximal promoter. Biochemical analyses identified hnRNP K as a specific protein that directly associates with the poly(C) motif in vitro. Interestingly, we further confirmed the rhythmic binding of endogenous hnRNP K within the Dbp promoter through chromatin immunoprecipitation as well as the cycling expression of hnRNP K. Finally, knockdown of hnRNP K decreased mRNA oscillation in both Dbp and Dbp-dependent clock genes. Taken together, our results show rhythmic protein expression of hnRNP K and provide new insights into its function as a transcriptional amplifier of Dbp.

scholarly journals Modulation of FoxO1 Expression by miR-21 to Promote Growth of Pancreatic Ductal Adenocarcinoma

2015 ◽  
Vol 35 (1) ◽  
pp. 184-190 ◽  
Author(s):  
Weifeng Song ◽  
Qi Li ◽  
Lei Wang ◽  
Liwei Wang
Keyword(s):  
Pancreatic Ductal Adenocarcinoma ◽  
Underlying Mechanism ◽  
Ductal Adenocarcinoma ◽  
Luciferase Reporter ◽  
Reporter Assay ◽  
Primary Tumors ◽  
Protein Levels ◽  
Infusion System

Background: Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal primary tumors in humans, with undetermined tumorigenesis. Although previous work by us, and by others, has clearly demonstrated an involvement of miR-21 in the growth of PDAC, the underlying mechanism has not been clarified. Methods: Here we analyzed the regulation of FoxO1 by miR-21 in vitro and in vivo, using luciferase-reporter assay and pancreatic intraductal infusion of antisense of miR-21, respectively. Results: We found that overexpression of miR-21 in PDAC cells decreased FoxO1 protein levels, whereas inhibition of miR-21 increased FoxO1 levels. Further, miR-21 bound to FoxO1 mRNA to prevent its translation through its 3'UTR. Moreover, administration of antisense of miR-21 through an intraductal infusion system significantly decreased miR-21 levels and increased FoxO1 levels in implanted PDAC, resulting in a significant decrease in PDAC growth. Conclusion: Taken together, our data highlight miR-21/FoxO1 axis as a novel therapeutic target for inhibiting the growth of PDAC.

scholarly journals Imipramine Influences Body Distribution of Supplemental Zinc Which May Enhance Antidepressant Action

Nutrients ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 2529
Author(s):  
Anna Rafało-Ulińska ◽  
Ewa Poleszak ◽  
Aleksandra Szopa ◽  
Anna Serefko ◽  
Magdalena Rogowska ◽  
...  
Keyword(s):  
Protein Level ◽  
Forced Swim Test ◽  
Underlying Mechanism ◽  
Intestinal Cell ◽  
Control Group ◽  
Protein Levels ◽  
Body Distribution ◽  
Antidepressant Efficacy ◽  
The Brain

Zinc (Zn) was found to enhance the antidepressant efficacy of imipramine (IMI) in human depression and animal tests/models of depression. However, the underlying mechanism for this effect remains unknown. We measured the effect of intragastric (p.o.) combined administration of IMI (60 mg/kg) and Zn (40 mg Zn/kg) in the forced swim test (FST) in mice. The effect of Zn + IMI on serum, brain, and intestinal Zn concentrations; Zn transporter (ZnT, ZIP) protein levels in the intestine and ZnT in the brain; including BDNF (brain-derived neurotrophic factor) and CREB (cAMP response element-binding protein) protein levels in the brain were evaluated. Finally, the effect of IMI on Zn permeability was measured in vitro in colon epithelial Caco-2 cells. The co-administration of IMI and Zn induced antidepressant-like activity in the FST in mice compared to controls and Zn or IMI given alone. This effect correlated with increased BDNF and the ratio of pCREB/CREB protein levels in the prefrontal cortex (PFC) compared to the control group. Zn + IMI co-treatment increased Zn concentrations in the serum and brain compared to the control group. However, in serum, co-administration of IMI and Zn decreased Zn concentration compared to Zn alone treatment. Also, there was a reduction in the Zn-induced enhancement of ZnT1 protein level in the small intestine. Zn + IMI also induced an increase in the ZnT4 protein level in the PFC compared to the control group and normalized the Zn-induced decrease in the ZnT1 protein level in the hippocampus (Hp). The in vitro studies revealed enhanced Zn permeability (observed as the increased transfer of Zn through the intestinal cell membrane) after IMI treatment. Our data indicate that IMI enhances Zn transfer through the intestinal tract and influences the redistribution of Zn between the blood and brain. These mechanisms might explain the enhanced antidepressant efficacy of combined IMI/Zn treatment observed in the FST in mice.

scholarly journals The fibroblast-derived protein PI16 controls neuropathic pain

2020 ◽  
Vol 117 (10) ◽  
pp. 5463-5471 ◽  
Author(s):  
Pooja Singhmar ◽  
Ronnie The Phong Trinh ◽  
Jiacheng Ma ◽  
XiaoJiao Huo ◽  
Bo Peng ◽  
...  
Keyword(s):  
Neuropathic Pain ◽  
Dorsal Root ◽  
Clinical Problem ◽  
Endothelial Barrier ◽  
Leukocyte Infiltration ◽  
Spared Nerve Injury ◽  
Protein Levels ◽  
Major Clinical Problem

Chronic pain is a major clinical problem of which the mechanisms are incompletely understood. Here, we describe the concept that PI16, a protein of unknown function mainly produced by fibroblasts, controls neuropathic pain. The spared nerve injury (SNI) model of neuropathic pain increases PI16 protein levels in fibroblasts in dorsal root ganglia (DRG) meninges and in the epi/perineurium of the sciatic nerve. We did not detect PI16 expression in neurons or glia in spinal cord, DRG, and nerve. Mice deficient in PI16 are protected against neuropathic pain. In vitro, PI16 promotes transendothelial leukocyte migration. In vivo, Pi16−/− mice show reduced endothelial barrier permeability, lower leukocyte infiltration and reduced activation of the endothelial barrier regulator MLCK, and reduced phosphorylation of its substrate MLC2 in response to SNI. In summary, our findings support a model in which PI16 promotes neuropathic pain by mediating a cross-talk between fibroblasts and the endothelial barrier leading to barrier opening, cellular influx, and increased pain. Its key role in neuropathic pain and its limited cellular and tissue distribution makes PI16 an attractive target for pain management.

scholarly journals MicroRNA-214-3p Inhibits the Stem-Like Properties of Lung Squamous Cell Cancer by Targeting YAP1

2020 ◽  
Author(s):  
Tingting Lu ◽  
Ying Yang ◽  
Ziming Li ◽  
Shun Lu
Keyword(s):  
Squamous Cell ◽  
Squamous Cell Cancer ◽  
Cell Cancer ◽  
Underlying Mechanism ◽  
Cell Counting ◽  
Hippo Signaling ◽  
Kaplan Meier ◽  
Luciferase Activity Assay

Abstract Background: Emerging evidence reveals that microRNAs (miRNAs) play a crucial role in tumor progression, but the underlying mechanism of microRNAs in lung squamous cell cancer (LSCC) remains unclear. Method : Western-blotting and quantitative real-time PCR (q-PCR) were carried out to detect mRNA and protein expression. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK-8), colony-forming assay or sphere-forming assay, respectively. Results: MiR-214-3p was markedly de-regulated in LSCC tissues and was inversely related to the level of Yes-associated protein1 (YAP1), which is the core transcription regulator of the Hippo signaling pathway. Kaplan-Meier survival curves illustrated that patients with high miR-214-3p expression demonstrated more favorable clinical outcomes. MiR-214-3p overexpression (OE) repressed proliferation and cancer stem-like cells (CSCs) properties in vitro and in vivo xenograft mouse model. Mechanistically, luciferase activity assay revealed that miR-214-3p directly targets YAP1 by specifically binding on the 3’ UTR of YAP1. Conclusion: MiR-214-3p plays a pivotal role in CSCs properties by targeting YAP1, which provides a potential treatment strategy for LSCC patients.

scholarly journals Exosomes from LPS-Stimulated hDPSCs Activated the Angiogenic Potential of HUVECs In Vitro

2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Xiangyu Huang ◽  
Wei Qiu ◽  
Yuhua Pan ◽  
Jianjia Li ◽  
Zhao Chen ◽  
...  
Keyword(s):  
Pathway Analysis ◽  
Target Genes ◽  
Underlying Mechanism ◽  
Tube Formation ◽  
Endothelial Cell Proliferation ◽  
Inflammatory Factor ◽  
Angiogenic Potential ◽  
Vascular Regeneration ◽  
Microrna Sequencing

Background. Exosomes from human dental pulp stem cells (hDPSCs) were indicated to play a positive role in vascular regeneration processes. But the angiogenic capabilities of exosomes from inflammatory hDPSCs and the underlying mechanism remain unknown. In this study, the inflammatory factor lipopolysaccharide (LPS) was used to stimulate hDPSCs, and exosomes were extracted from these hDPSCs. The proangiogenic potential of exosomes was examined, and the underlying mechanism was studied. Method. Exosomes were isolated from hDPSCs with or without LPS stimulation (N-EXO and LPS-EXO) and cocultured with human umbilical vein endothelial cells (HUVECs). The proangiogenic potential of exosomes was evaluated by endothelial cell proliferation, migration, and tube formation abilities in vitro. To investigate the proangiogenic mechanism of LPS-EXO, microRNA sequencing was performed to explore the microRNA profile of N-EXO and LPS-EXO. Gene Ontology (GO) analysis was used to study the functions of the predicted target genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was used to estimate the signaling pathways associated with the inflammation-induced angiogenesis process. Result. Compared to the uptake of N-EXO, uptake of LPS-EXO activated the angiogenic potential of HUVECs by promoting the proliferation, migration, and tube formation abilities in vitro. The mRNA expression levels of vascular endothelial growth factor (VEGF) and kinase-insert domain-containing receptor (KDR) in the LPS-EXO group were significantly higher than those in the N-EXO group. MicroRNA sequencing showed that 10 microRNAs were significantly changed in LPS-EXO. Pathway analysis showed that the genes targeted by differentially expressed microRNAs were involved in multiple angiogenesis-related pathways. Conclusion. This study revealed that exosomes derived from inflammatory hDPSCs possessed better proangiogenic potential in vitro. This is the first time to explore the role of exosomal microRNA from hDPSCs in inflammation-induced angiogenesis. This finding sheds new light on the effect of inflammation-stimulated hDPSCs on tissue regeneration.

scholarly journals Rehin Mitigates Chronic Kidney Injury by Decreasing the Release of Perforin and IFN-γ from Immune Cells Via Inhibiting the STING/TBK1/IRF3 Pathway Activation

2022 ◽  
Author(s):  
Jingyu Wang ◽  
Xin Huang ◽  
Yong Liao ◽  
Xintian Cai ◽  
Jing Xu ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Immune Cells ◽  
Medicinal Preparation ◽  
Kidney Injury ◽  
Normal Subjects ◽  
Inflammatory Factor ◽  
Mice Model ◽  
Qrt Pcr ◽  
Ifn Γ

Abstract Shenkang suppository (SKS), a Chinese medicinal preparation rich in various natural ingredients, has not been reported in any studies related to fibrosis. Our experiments validated the anti-fibrotic and anti-inflammatory effects of Rhein (Rh), which is a major component of SKS, and explored its potential immune mechanisms. Tissue and serum specimens from chronic kidney disease (CKD) patients and normal subjects were collected in 30 cases each, and the expression differences of perforin and IFN-γ were analyzed by ELISA. Further, the CKD mice model constructed with folic acid (FA) was used to validate these differences by WB and qRT-PCR to explore the potential nephroprotective mechanism of Rh. Besides, in vitro experiments were conducted to identify the release sources of perforin and IFN-γ. ELISA showed that perforin and IFN-γ were upregulated in CKD patients, and this phenomenon was also corroborated in CKD mice. WB and qRT-PCR data showed that Rh reversed perforin and IFN-γ upregulation, inflammatory factor recruitment, and extracellular matrix (ECM) protein upregulation. Results from in vitro experiments demonstrate that the upregulation of perforin and IFN-γ originates from the stress response of CD4+ T lymphocytes (CD4+ cells), CD8+ T lymphocytes (CD8+ cells) and natural killer cells (NK cells), which can be suppressed by Rh. More importantly, the activated STING/TBK1/IRF3 pathway in CKD was also inhibited by Rh. Our data suggest that Rh possesses anti-fibrotic and nephroprotective effects, which mechanistically are associated with decreased release of perforin and IFN-γ from immune cells, which may be achieved by suppressing the STING/TBK1/IRF3 pathway.

Di-2-ethylhexyl phthalate (DEHP) induces apoptosis of mouse HT22 hippocampal neuronal cells via oxidative stress

2020 ◽  
Vol 36 (11) ◽  
pp. 844-851
Author(s):  
Wei Tu ◽  
Weifeng Li ◽  
Xingen Zhu ◽  
Linlin Xu
Keyword(s):  
Oxidative Stress ◽  
Cell Viability ◽  
Neuronal Cell ◽  
Underlying Mechanism ◽  
Treated Group ◽  
Dependent Manner ◽  
Ht22 Cells ◽  
Protein Levels ◽  
Ethylhexyl Phthalate

Di-2-ethylhexyl phthalate (DEHP) has been widely used as a plasticizer in industry and can affect memory; however, the underlying mechanism remains unclear. In the present study, mouse HT22 cells, an immortalized hippocampal neuronal cell line, was utilized as an in vitro model. We showed that DEHP dramatically inhibited cell viability and increased lactate dehydrogenase (LDH) release from the cells in a dose-dependent manner, suggesting that DEHP could cause cytotoxicity of mouse HT22 cells. The protein levels of cleaved Caspase-8, cleaved Caspase-3, and Bax markedly increased in the DEHP-treated cells, whereas there was a significant decrease in the Bcl-2 protein level, implying that DEHP could induce apoptosis of mouse HT22 cells. DEHP exposure significantly increased the content of malondialdehyde, whereas it markedly decreased the level of glutathione and the activities of glutathione peroxidase and superoxide dismutase, suggesting that DEHP induced oxidative stress of the cells. Compared with the DEHP-treated group, the inhibition of cell viability and the release of LDH were rescued in the N-acetyl-l-cysteine plus DEHP group. Furthermore, inhibition of oxidative stress could rescue the induction of apoptosis by DEHP. Collectively, our results indicated that DEHP could induce apoptosis of mouse HT22 cells via oxidative stress.

scholarly journals Fn14 Exacerbates Acute Lung Injury by Activating the NLRP3 Inflammasome in Mice

2021 ◽  
Author(s):  
Xin-Xin Guan ◽  
Hui-Hui Yang ◽  
Wen-Jing Zhong ◽  
Jia-Xi Duan ◽  
Chen-Yu Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Acute Lung Injury ◽  
Survival Rate ◽  
Lung Injury ◽  
Nlrp3 Inflammasome ◽  
Inflammasome Activation ◽  
Inflammatory Factor ◽  
Caspase 1 ◽  
Primary Mouse

Abstract Background: Uncontrolled inflammation is an important factor in the occurrence and development of acute lung injury (ALI). Fibroblast growth factor-inducible 14 (Fn14) takes part in the pathological process of a variety of inflammatory diseases. However, the role of Fn14 in ALI has not yet been elucidated. Methods: C57BL/6J mice were used in this study. ALI model was induced by intratracheal injection of lipopolysaccharide (LPS, 5 mg/kg). The effects of Fn14 receptor blocker ATA (20 mg/kg) on lung injury, inflammatory cell infiltration, inflammatory factor secretion, and oxidative stress in mice were observed. The activation of NLRP3 inflammasome was detected by qPCR, Western blot, and ELISA. Prophylactic or therapeutic ATA was administered to observe its effect on the survival rate of ALI mice. In vitro, primary mouse peritoneal macrophages were used to activate the NLRP3 inflammasome by LPS or LPS+ATP. Fn14 was activated by recombinant TWEAK, or knockdown by lentivirus, and the effects on NLRP3 inflammasome activation was detected.Results: We found that ATA significantly downregulated the expression of Fn14 in the lungs and improved the survival rate of mice receiving a lethal dose of LPS. ATA also attenuated lung tissue damage by decreasing the infiltration of macrophages and neutrophils, reducing inflammation, and suppressing oxidative stress. Interestingly, we found that ATA strongly inhibited the activation of NLRP3 inflammasome in the lungs of ALI mice. Furthermore, in vitro, exogenous TWEAK, a natural ligand of Fn14, enhanced the levels of NLRP3 and Caspase-1 p10 and the maturation and secretion of IL-1β in the primary murine macrophages, eventually leading to the activation of NLRP3 inflammasome. In addition, the expression of Fn14, NLRP3, and Caspase-1 p10 and the production of IL-1β were effectively blocked by Fn14 shRNA in macrophages. In mechanism, the activation of Fn14 promoted the production of reactive oxygen species in activated macrophages. Conclusion：Our study first reports that the activation of Fn14 aggravates ALI by amplifying the activation of NLRP3 inflammasome. Therefore, blocking Fn14 may be a potential way to treat ALI.

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