scholarly journals miRNA-544a Regulates the Inflammation of Spinal Cord Injury by Inhibiting the Expression of NEUROD4

2018 ◽  
Vol 51 (4) ◽  
pp. 1921-1931 ◽  
Author(s):  
Lei Yang ◽  
Dawei Ge ◽  
Xi Chen ◽  
Chunzhi Jiang ◽  
Shengnai Zheng
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Grip Strength ◽  
Target Gene ◽  
Interleukin 1 ◽  
Luciferase Reporter ◽  
Interleukin 17 ◽  
Qrt Pcr ◽  
Gene Assay ◽  
Cord Injury

Background/Aims: To explore the potential role of miR-544a in spinal cord injury and the possible mechanism involved. Methods: We established a mouse model with spinal cord injury to examine the changes in grip force recovery of the forelimb or the posterior limb of the mouse. Microarray was performed to achieve differentiated miRNAs in the mice. The expressions of miR-544a, MCP-1, IL36B and IL17B after spinal cord injury were detected by qRT-PCR. Subsequently, miR-544a was overexpressed to observe changes in inflammation and grip strength after spinal cord injury. Target gene of miR-544a was then predicted using bioinformatics technology. Finally, dual luciferase reporter gene assay was used to verify the binding of miR-544a to its target gene. Results: Using mice models with spinal cord injury, we found that the strength of their four limbs began to recover 7 days after injury. The results of microarray and qRT-PCR confirmed that mir-544a level in mice with spinal cord injury decreased with increase of injury time, while the levels of inflammatory genes MCP-1 (monocyte chemoattractant protein-1), IL1 (interleukin-1) and TNF-α (tumor necrosis factor alpha) IL36B (interleukin-36 beta) and IL17B (interleukin-17 beta) were significantly increased. However, overexpression of miR-544a in the mice significantly reduced the level of inflammation and restored their grip strength in their four limbs. Finally, we found that miR-544a can bind to the NEUROD4 (Neurogenic differentiation 4) 3’UTR (Untranslated Region) region through bioinformatics website prediction, which was further confirmed by dual luciferase reporter assay. NEUROD4 level was significantly reduced following the overexpression of miR-544a. Conclusion: The expression of miR-544a was significantly decreased after spinal cord injury. High expression of miR-544a could alleviate the inflammation caused by spinal cord injury and promote the recovery of spinal cord via the inhibition of NEUROD4.

scholarly journals mir-21a-5p promote the progress of inflammation after Traumatic Spinal Cord Injury via up-regulating neurotoxic reactive astrocyte (A1) polarization by inhibiting CNTF/STAT3/Nkrf pathway

2020 ◽  
Author(s):  
Yining Zhang ◽  
Tingting Meng ◽  
Jianan Chen ◽  
Ying Zhang ◽  
Jianning Kang ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Reactive Astrocytes ◽  
Luciferase Reporter ◽  
Chip Assay ◽  
Qrt Pcr ◽  
Pulldown Assay ◽  
Cord Injury

Abstract Background Reactive astrocytes play an important role in Traumatic Spinal Cord Injury (TSCI). Interestingly, naive astrocytes can easily transform into neurotoxic reactive astrocytes(A1s) when inflammatory stimulation occurs. Previous researches have reported that miR-21a-5p is involved in the regulation of various stages of Spinal Cord Injury (SCI). However, it is not clear whether miR-21a-5p affected the polarization of reactive astrocytes. The purpose of our study was to detect the effects and mechanism of miR-21a-5p in the induction of neurotoxic reactive astrocytes (A1s) formation. Methods Gene chip assay and qRT-PCR were used to detect the expression of Cntfr α in TSCI models or sham operation. Bioinformatics analysis was used to speculate the potential targeting of miR-21a-5p, which was further confirmed by qRT-PCR, western blotting, a dual-luciferase reporter assay, and RNA pulldown assay. In vivo, the TSCI model was performed by a 68099Ⅱ precision percussion device, and the A1s phenotype was identified by immunofluorescence staining. In vitro, A1s were induced by IL-1 α, TNF-α, and C1q. A1s and neuroprotective reactive astrocytes (A2s) markers were confirmed by qRT-PCR, western blotting, and immunofluorescence. ChIP assay was used to explore the targeting gene of STAT3, the downstream of Cntfr α. Results The expression of miR-21a-5p was significantly increased while Cntfr α was decreased since naive astrocytes transformed into A1s after 3 days post-TSCI. In addition, the mRNA and protein of Cntfr α were decreased while miR-21a-5p was overexpressed. The binding site between miR-21a-5p and Cntfr α was further confirmed by the dual-luciferase reporter and RNA pulldown assay. We also discovered that A1s markers were decreased while markers of A2s were increased with the pretreatment of CNTF. Chromatin immunoprecipitation (ChIP) assay was used to prove that CNTF inhibited A1s induction by activating the expression of Nkrf via the CNTF/STAT3 pathway. Downregulation of miR-21a-5p enhanced the inhibitory effect of CNTF in A1s in vitro. In vivo, the expression of A1s markers significantly decreased with the treatment of antagomir-21, while Cntfr α siRNA treatment was just the opposite. Conclusion We observed that increased miR-21a-5p down-regulated Cntfr α in A1s induced by TSCI, promoting the inflammatory process. In addition, we also identified the effect and potential mechanism of CNTF, a specific ligand of CNTFR α, on inhibiting naive astrocytes transformed into A1s for the first time. Collectively, our studies demonstrated that targeting miR-21a-5p is a prospective therapy for curing TSCI.

scholarly journals Hsa-miR-105-1 Regulates Cisplatin-Resistance in Ovarian Carcinoma Cells by Targeting ANXA9

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Xinxin Kou ◽  
Hui Ding ◽  
Lei Li ◽  
Hongtu Chao
Keyword(s):  
Ovarian Carcinoma ◽  
Cell Lines ◽  
Target Gene ◽  
Molecular Mechanisms ◽  
Cisplatin Resistance ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Qrt Pcr ◽  
Gene Assay ◽  
Increased Sensitivity

Purpose. Cisplatin is one of the most effective drugs for treating ovarian carcinoma (OC), which is among the most lethal types of carcinoma. However, the chemoresistance to cisplatin that develops over time leads to a poor clinical outcome for many OC patients. Therefore, it is necessary to clearly understand the molecular mechanisms of chemoresistance. In this study, we examined how Hsa-miR-105-1 functions in cisplatin-resistant OC cells. Methods. The levels of Hsa-miR-105-1 expression in cisplatin-sensitive and resistant OC cell lines were detected by qRT-PCR. The target gene of Hsa-miR-105-1 was predicted by using the TargetScan and Starbase databases and verified by the double luciferase reporter gene assay. The target gene of Hsa-miR-105-1 was identified as ANXA9, and ANXA9 expression was evaluated by qRT-PCR, western blotting, and immunofluorescence. To validate the function of Hsa-miR-105-1 in OC cells, we silenced or overexpressed Hsa-miR-105-1 in cisplatin-sensitive or resistant OC cell lines, respectively. Furthermore, the expression levels of several apoptosis-related proteins, including P53, P21, E2F1, Bcl-2, Bax, and caspase-3, were examined by western blot analysis. Results. The levels of Hsa-miR-105-1 expression were abnormally downregulated in cisplatin-resistant OC cells, while ANXA9 expression was significantly upregulated in those cells. Treatment with an Hsa-miR-105-1 inhibitor promoted the expression of ANXA9 mRNA and protein, enhanced the resistance to cisplatin, and attenuated the cell apoptosis induced by cisplatin in cisplatin-sensitive OC cells. Moreover, treatment with Hsa-miR-105-1 mimics inhibited ANXA9 expression, which further increased the levels of P53, P21, and Bax expression and decreased the levels of E2F1 and Bcl-2 expression, finally resulting in an increased sensitivity to cisplatin in cisplatin-resistant OC cells. Conclusion. We found that a downregulation of Hsa-miR-105-1 expression enhanced cisplatin-resistance, while an upregulation of Hsa-miR-105-1 restored the sensitivity of OC cells to cisplatin. The Hsa-miR-105-1/ANXA9 axis plays an important role in the cisplatin-resistance of OC cells.

scholarly journals Inhibiting HMGB1-RAGE axis prevents pro-inflammatory macrophages/microglia polarization and affords neuroprotection after spinal cord injury

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Hong Fan ◽  
Hai-Bin Tang ◽  
Zhe Chen ◽  
Hu-Qing Wang ◽  
Lei Zhang ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Western Blot ◽  
Functional Recovery ◽  
Sterile Inflammation ◽  
Qrt Pcr ◽  
Cord Injury ◽  
Microglia Polarization ◽  
Anti Inflammatory ◽  
Inflammatory Macrophages

Abstract Background Spinal cord injury (SCI) favors a persistent pro-inflammatory macrophages/microglia-mediated response with only a transient appearance of anti-inflammatory phenotype of immune cells. However, the mechanisms controlling this special sterile inflammation after SCI are still not fully elucidated. It is known that damage-associated molecular patterns (DAMPs) released from necrotic cells after injury can trigger severe inflammation. High mobility group box 1(HMGB1), a ubiquitously expressed DNA binding protein, is an identified DAMP, and our previous study demonstrated that reactive astrocytes could undergo necroptosis and release HMGB1 after SCI in mice. The present study aimed to explore the effects and the possible mechanism of HMGB1on macrophages/microglia polarization, as well as the neuroprotective effects by HMGB1 inhibition after SCI. Methods In this study, the expression and the concentration of HMGB1 was determined by qRT-PCR, ELISA, and immunohistochemistry. Glycyrrhizin was applied to inhibit HMGB1, while FPS-ZM1 to suppress receptor for advanced glycation end products (RAGE). The polarization of macrophages/microglia in vitro and in vivo was detected by qRT-PCR, immunostaining, and western blot. The lesion area was detected by GFAP staining, while neuronal survival was examined by Nissl staining. Luxol fast blue (LFB) staining, DAB staining, and western blot were adopted to evaluate the myelin loss. Basso-Beattie-Bresnahan (BBB) scoring and rump-height Index (RHI) assay was applied to evaluate locomotor functional recovery. Results Our data showed that HMGB1 can be elevated and released from necroptotic astrocytes and HMGB1 could induce pro-inflammatory microglia through the RAGE-nuclear factor-kappa B (NF-κB) pathway. We further demonstrated that inhibiting HMGB1 or RAGE effectively decreased the numbers of detrimental pro-inflammatory macrophages/microglia while increased anti-inflammatory cells after SCI. Furthermore, our data showed that inhibiting HMGB1 or RAGE significantly decreased neuronal loss and demyelination, and improved functional recovery after SCI. Conclusions The data implicated that HMGB1-RAGE axis contributed to the dominant pro-inflammatory macrophages/microglia-mediated pro-inflammatory response, and inhibiting this pathway afforded neuroprotection for SCI. Thus, therapies designed to modulate immune microenvironment based on this cascade might be a prospective treatment for SCI.

scholarly journals Synthesis and Characterization of a Silica-Based Drug Delivery System for Spinal Cord Injury Therapy

2019 ◽  
Vol 11 (1) ◽  
Author(s):  
Guodong Sun ◽  
Shenghui Zeng ◽  
Xu Liu ◽  
Haishan Shi ◽  
Renwen Zhang ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Mesoporous Silica ◽  
Drug Delivery System ◽  
Delivery System ◽  
Central Component ◽  
High Dose ◽  
Interleukin 17 ◽  
Cord Injury

Abstract Acute inflammation is a central component in the progression of spinal cord injury (SCI). Anti-inflammatory drugs used in the clinic are often administered systemically at high doses, which can paradoxically increase inflammation and result in drug toxicity. A cluster-like mesoporous silica/arctigenin/CAQK composite (MSN-FC@ARC-G) drug delivery system was designed to avoid systemic side effects of high-dose therapy by enabling site-specific drug delivery to the spinal cord. In this nanosystem, mesoporous silica was modified with the FITC fluorescent molecule and CAQK peptides that target brain injury and SCI sites. The size of the nanocarrier was kept at approximately 100 nm to enable penetration of the blood–brain barrier. Arctigenin, a Chinese herbal medicine, was loaded into the nanosystem to reduce inflammation. The in vivo results showed that MSN-FC@ARC-G could attenuate inflammation at the injury site. Behavior and morphology experiments suggested that MSN-FC@ARC-G could diminish local microenvironment damage, especially reducing the expression of interleukin-17 (IL-17) and IL-17-related inflammatory factors, inhibiting the activation of astrocytes, thus protecting neurons and accelerating the recovery of SCI. Our study demonstrated that this novel, silica-based drug delivery system has promising potential for clinical application in SCI therapy.

Upregulation of type I interleukin−1 receptor after traumatic spinal cord injury in adult rats

2006 ◽  
Vol 111 (3) ◽  
pp. 220-228 ◽  
Author(s):  
Xiao-Fei Wang ◽  
Li-Dong Huang ◽  
Pan-Pan Yu ◽  
Jian-Guo Hu ◽  
Lan Yin ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Interleukin 1 ◽  
Traumatic Spinal Cord Injury ◽  
Type I ◽  
Adult Rats ◽  
Cord Injury

MicroRNA-448 targets SATB1 to reverse the cisplatin resistance in lung cancer via mediating Wnt/β-catenin signalling pathway

2020 ◽  
Vol 168 (1) ◽  
pp. 41-51
Author(s):  
Mei-Ying Ning ◽  
Zhao-Lin Cheng ◽  
Jing Zhao
Keyword(s):  
Lung Cancer ◽  
Target Gene ◽  
A549 Cells ◽  
Resistance Index ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Luciferase Reporter Gene Assay ◽  
Lung Cancer Patients ◽  
Qrt Pcr ◽  
Gene Assay

Abstract This study aims to examine whether miR-448 reverses the cisplatin (DDP) resistance in lung cancer by modulating SATB1. QRT-PCR and immunohistochemistry were used to examine the miR-448 and SATB1 expressions in DDP-sensitive and -resistant lung cancer patients. A microarray was used to investigate the cytoplasmic/nucleic ratio (C/N ratios) of genes in A549 cells targeted by miR-448, followed by Dual-luciferase reporter gene assay. A549/DDP cells were transfected with miR-448 mimics/inhibitors with or without SATB1 siRNA followed by MTT assay, Edu staining, flow cytometry, qRT-PCR and western blotting. MiR-448 was lower but SATB1 was increased in DDP-resistant patients and A549/DDP cells. And the patients showed low miR-448 expression or SATB1 positive expression had poor prognosis. SATB1, as a target gene with higher C/N ratios (>1), was found negatively regulated by miR-448. Besides, miR-448 inhibitors increased resistance index of A549/DDP cells, promoted cell proliferation, increased cell distribution in S phrase, declined cell apoptosis and activated Wnt/β-catenin pathway. However, SATB1 siRNA could reverse the above effect caused by miR-448 inhibitors. MiR-448 targeting SATB1 to counteract the DDP resistance of lung cancer cells via Wnt/β-catenin pathway.

scholarly journals Possible Involvement of Interleukin-1 in Cyclooxygenase-2Induction After Spinal Cord Injury in Rats

1999 ◽  
Vol 72 (1) ◽  
pp. 302-309 ◽  
Author(s):  
Takeharu Tonai ◽  
Yutaka Taketani ◽  
Natsuo Ueda ◽  
Takehiko Nishisho ◽  
Yasukazu Ohmoto ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Interleukin 1 ◽  
Cord Injury

scholarly journals Resveratrol Activated Sonic Hedgehog Signaling to Enhance Viability of NIH3T3 Cells in Vitro via Regulation of Sirt1

2018 ◽  
Vol 50 (4) ◽  
pp. 1346-1360 ◽  
Author(s):  
Shuang Guo ◽  
Hongyan Liao ◽  
Jie Liu ◽  
Jing Liu ◽  
Fanren Tang ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Hedgehog Signaling ◽  
Transcriptional Activity ◽  
Luciferase Reporter ◽  
Shh Signaling ◽  
Nih3t3 Cells ◽  
Cord Injury ◽  
Fibrotic Scar ◽  
Gli 1

Background/Aims: Injuries of the brain and spinal cord result in the formation of glial (reactive gliosis) and fibrotic (formed by fibroblasts) scars. Recent studies have shown that the fibrotic scar was much more important for hindering regeneration after brain or spinal cord injury than the astrocytic scar. However, it has been given much less attention for effects and mechanism of fibroblasts during formation of the fibrotic scar. Resveratrol may be a potential anti-scarring agent in burn-related scarring and keloid fibroblasts. However, it is unclear whether and how resveratrol affects formation of the fibrotic scar after brain or spinal cord injury. Earlier studies have shown that the activated Shh signaling has anti-apoptosis, anti-oxidation, anti-inflammation properties. Moreover, resveratrol can activate the Shh signaling. However, it is unclear how resveratrol activates the Shh signaling. Resveratrol is a activator of Sirt1. It is unknown whether resveratrol activates the Shh signaling via Sirt1. Methods: NIH3T3 cells, a fibroblast cell line, were used as model cells and treated with drugs. Cell viability was assessed by Cell Counting Kit 8. The expressions and activity of Shh signaling pathway proteins were evaluated by immunocytochemistry and Western blotting. Transcriptional activity of Gli-1 was detected with Dual-Luciferase Reporter Gene Assay Kit. Results: Resveratrol, Sirt1 agonist STR1720 and recombinant mouse Shh protein, an activator of hedgehog signaling, enhanced the viability of NIH3T3 cells, promoted Smo to translocated to the primary cilia and Gli-1 entered into the nuclei from cytoplasm, and upregulated expressions of Shh, Ptc-1, Smo, and Gli-1 proteins, which can be reversed by Smo antagonist cyclopamine and Sirt1 antagonist Sirtinol. Additionally, resveratrol increased transcriptional activity of Gli-1. Conclusion: We indicate in the first time that it may be mediated by Sirt1 for resveratrol activating the Shh signaling to enhance viability of NIH3T3 cells, and Sirt1 may be a regulator for upstream of the Shh signaling pathway.This study provides a basis for further investigating effects and mechanism of resveratrol during the formation of fibrous scar after brain or spinal cord injury.

Sign in / Sign up

Export Citation Format

Share Document