scholarly journals SHED-Derived Exosomes Regulate Microglial Polarization in the Treatment of Traumatic Brain Injury in Rats via miR-330-5p

2020 ◽  
Author(s):  
Ye Li ◽  
Xinxin Wang ◽  
Xiaoyu Cao ◽  
Na Li ◽  
Sun Meng ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Structural Damage ◽  
Inhibitory Effect ◽  
Deciduous Teeth ◽  
Luciferase Reporter ◽  
Enzyme Linked Immunosorbent Assay ◽  
M2 Polarization ◽  
M1 Polarization

Abstract Background: Traumatic brain injury (TBI) causes structural damage and impairs motor and cognitive function of the brain. Our previous study suggested that exosomes (EXs) secreted by stem cells from human exfoliated deciduous teeth (SHED) extenuated motor damage in TBI rats by regulating microglia. The molecular mechanism of SHED-EXs was investigated in the present study. Methods: The miRNA array was performed to determine the differential miRNA expression in SHED-EXs treating microglia. The key miRNA was selected. Flow cytometry, immunofluorescence, enzyme linked immunosorbent assay (ELISA) and Griess assay were performed to detect the function of key miRNA. Real-time PCR, Western blotting and dual luciferase reporter assay were used to confirm the relationship between key miRNA and the target gene. Chromatin immunoprecipitation (ChIP) was performed to determine the downstream pathway of EXs-miRNA. Traumatic brain injury rat model was established and local injection of EXs-miRNA was performed to evaluate the effect.Results: SHED-EXs delivery of miR-330-5p was the key in the regulation of microglia polarization by inhibiting M1 polarization and promoting M2 polarization. Mechanistically, miR-330-5p had an inhibitory effect on Ehmt2, and miR-330-5p/Ehmt2 promoted the transcription of CXCL14 through H3K9me2. In vivo data showed that SHED-EXs/miR-330-5p reduced neuro-inflammation and repaired neurological function of TBI rats. Conclusions: SHED-EXs/miR-330-5p improved the motor function of rats after TBI by inhibiting M1 polarization and promoting M2 polarization of microglia through Ehmt2/H3K9me2/CXCL14 pathway.

scholarly journals Electroacupuncture Improved Hippocampal Neurogenesis following Traumatic Brain Injury in Mice through Inhibition of TLR4 Signaling Pathway

2017 ◽  
Vol 2017 ◽  
pp. 1-13 ◽  
Author(s):  
Yuqin Ye ◽  
Yongxiang Yang ◽  
Chen Chen ◽  
Ze Li ◽  
Yanfeng Jia ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Signaling Pathway ◽  
Neurological Diseases ◽  
Inhibitory Effect ◽  
Protective Role ◽  
Hippocampal Neurogenesis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Tlr4 Signaling ◽  
Tlr4 Signaling Pathway

The protective role of electroacupuncture (EA) treatment in diverse neurological diseases such as ischemic stroke is well acknowledged. However, whether and how EA act on hippocampal neurogenesis following traumatic brain injury (TBI) remains poorly understood. This study aims to investigate the effect of EA on hippocampal neurogenesis and neurological functions, as well as its underlying association with toll-like receptor 4 (TLR4) signaling in TBI mice. BrdU/NeuN immunofluorescence was performed to label newborn neurons in the hippocampus after EA treatment. Water maze test and neurological severity score were used to evaluate neurological function posttrauma. The hippocampal level of TLR4 and downstream molecules and inflammatory cytokines were, respectively, detected by Western blot and enzyme-linked immunosorbent assay. EA enhanced hippocampal neurogenesis and inhibited TLR4 expression at 21, 28, and 35 days after TBI, but the beneficial effects of EA on posttraumatic neurogenesis and neurological functions were attenuated by lipopolysaccharide-induced TLR4 activation. In addition, EA exerted an inhibitory effect on both TLR4/Myd88/NF-κB and TLR4/TRIF/NF-κB pathways, as well as the inflammatory cytokine expression in the hippocampus following TBI. In conclusion, EA promoted hippocampal neurogenesis and neurological recovery through inhibition of TLR4 signaling pathway posttrauma, which may be a potential approach to improve the outcome of TBI.

scholarly journals Improved Promotion of M2 Microglial Polarization by Zuogui Jiangtang Jieyu Formulation in Diabetes-Related Depression

2021 ◽  
Author(s):  
xiuli zhang ◽  
Dahua Wu ◽  
Dandan Li ◽  
Jian Liu ◽  
Chang Lei ◽  
...  
Keyword(s):  
Inflammatory Cytokines ◽  
High Glucose ◽  
Neuroprotective Effect ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mild Stress ◽  
Microglial Polarization ◽  
M2 Polarization ◽  
M1 Polarization

Abstract Background Zuogui Jiangtang Jieyu formulation (ZGJTJY) is a Chinese polyherbal prescription for diabetes-related depression (DD). The mechanism underlying hippocampal M1/M2 polarization in DD and the ZGJTJY treatment effects remain unclear. This study aimed to investigate M1/M2 microglial polarization in the hippocampus of DD rats and HAPI (highly aggressively proliferating immortalized) cells simulating the DD state, as well as to examine the ZGJTJY intervention effects, both in vivo and in vitro. Methods We subjected Sprague Dawley rats to a high-fat diet, streptozotocin, and unpredictable chronic mild stress; subsequently, we orally administered ZGJTJY. HAPI cells were induced using high glucose and corticosterone; subsequently, ZGJTJY-containing serum was added to examine changes in M1/M2 microglial polarization. Moreover, metformin combined with fluoxetine (DMGB/F) was used as a positive drug for evaluating the ZGJTJY intervention. Laser confocal scanning was used to examine the microglial morphology. Further, real-time PCR was used to determine M1 markers (MHCII, iNOS, MCP-1, CD11b), M2 markers (Arg1, Mrc1, Ym1), pro-inflammatory cytokines (IL-1β, IL-6, TNF-α), and anti-inflammatory cytokines (IL-4, IL-10). Additionally, an enzyme-linked immunosorbent assay was used to examine inflammatory cytokines. Results There was significant activation of M1 polarization in the hippocampus of DD rats and HAPI cells induced using high glucose and corticosterone. Compared with DMGB/F, ZGJTJY inhibited and promoted M1 and M2 polarization, respectively; moreover, it decreased the M1-to-M2 polarization ratio both in vivo and in vitro. Conclusions The study indicated that hippocampal M1 polarization is crucially involved in DD pathogenesis; moreover, there is a need for further research on the neuroprotective effect of Chinese medicine associated with M2-polarized microglia.

Manual acupuncture relieves microglia-mediated neuroinflammation in a rat model of traumatic brain injury by inhibiting the RhoA/ROCK2 pathway

2020 ◽  
Vol 38 (6) ◽  
pp. 426-434
Author(s):  
Ming-min Zhu ◽  
Ji-huan Lin ◽  
Peng Qing ◽  
Liu Pu ◽  
Shu-lian Chen ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Signaling Pathway ◽  
Inflammatory Factors ◽  
Enzyme Linked Immunosorbent Assay ◽  
Manual Acupuncture ◽  
Sprague Dawley ◽  
Tissue Samples ◽  
M1 Polarization ◽  
Immunofluorescence Labeling

Objective: To investigate the regulatory mechanism of manual acupuncture (MA) on microglial polarization–mediated neuroinflammation after traumatic brain injury (TBI), focusing on the RhoA/Rho-associated coiled coil-forming protein kinase (ROCK2) pathway. Methods: Sprague Dawley (SD) rats were used to generate a TBI model using Feeney’s freefall epidural impact method. MA was performed on half of the TBI model rats, while the others remained untreated. Acupuncture was administered at GV15, GV16, GV20, GV26, and LI4. At the end of the intervention, rat brain tissue samples were collected, and the microglial M1 polarization status was observed by immunofluorescence labeling of CD86, an M1 microglia-specific protein. RhoA/ROCK2 signaling components were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. An enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of inflammatory factors. Results: Compared with normal rats, the CD86 expression density in the untreated TBI model rats was high and showed an aggregated expression pattern. The genes and proteins of the RhoA/ROCK2 signaling pathway were highly expressed, and inflammatory factors were significantly increased. The CD86 expression density in TBI rats after MA was reduced compared to that in untreated TBI rats and showed a scattered distribution. The expression of RhoA/ROCK2 signaling pathway genes and proteins was also significantly reduced, and inflammatory factors were decreased. Conclusion: These results show that MA may inhibit M1 polarization of microglia by regulating the RhoA/ROCK2 signaling pathway, thereby reducing neuroinflammation in TBI.

scholarly journals Extracellular vesicles-transmitted miR-21a-5p altered microglia polarization after hypoxia-ischemic injury in neonatal mice via STAT3 pathways

2021 ◽  
Author(s):  
Danqing Xin ◽  
Yijing Zhao ◽  
Tingting Li ◽  
Hongfei Ke ◽  
Chengcheng Gai ◽  
...  
Keyword(s):  
Brain Injury ◽  
Brain Damage ◽  
Extracellular Vesicles ◽  
Luciferase Reporter ◽  
Neonatal Mice ◽  
M2 Polarization ◽  
Stat3 Phosphorylation ◽  
Microglia Polarization

Abstract Background We previously reported that mesenchymal stromal cells (MSCs)-derived extracellular vesicles (EVs) exhibit protective effects in hypoxia-ischemia (HI) brain damage. The neuroprotective action was connected with its anti-inflammatory effect. However, the mechanisms involved with this effect have not been determined. Methods A modified version of the Rice-Vannucci method was performed on postnatal day 7 mouse pups to induce neonatal HI brain injury. The model of oxygen-glucose deprivation (OGD) was established in BV-2 cells to mimic HI injury in vitro. Mice or BV-2 cells received EVs and EVs-miR-21ainhibitor at indicative time post-injury. In vivo, brain water content and TTC staining were used to evaluate the effects of EVs on HI brain injury. Immunofluorescence staining was used to observe the effect of EVs on the polarization of microglia. The effect of EVs on p-STAT3 was assessed by Western blot. In vitro, the effect of EVs on cell survival was evaluated by CCK8. Expression of miR-21a-5p and inflammatory factors was measured using qRT-PCR. Dual-Luciferase Reporter Assay was performed to illustrate the link between miR-21a-5p and STAT3. The role of miR-21a-5p in EVs on HI injury and ODG injury was further investigated by using EVs-miR-21ainhibitor.Results By using OGD mimicking HI injury in vitro, we found that MSCs-EVs treatment elevated cell viability following OGD exposure in BV-2 cells. MSCs-EVs treatment impeded microglia-mediated neuroinflammation, shifted microglia toward M2 polarization, and suppressed the phosphorylation of selective signal transducer and activator of transcription 3 (STAT3) in microglia after HI exposure in vitro and in vivo. In light of miR-21a-5p being the most highly expressed miRNA in MSCs-EVs interacting with the STAT3 pathway, further work focused on this pathway. Notably, MSCs-EVs treatment increased HI-reduced miR-21a-5p levels in BV-2 cells. Diminishing miR-21a-5p in MSCs-EVs partially attenuated its effect on microglia polarization and STAT3 phosphorylation following HI exposure in vitro and in vivo. Conclusions Our study suggested that MSCs-EVs attenuated HI brain damage in neonatal mice via shuttling miR-21a-5p, which induced microglia M2 polarization by targeting STAT3.

Changqin NO. 1 inhibits neuronal apoptosis via suppressing GAS5 expression in a traumatic brain injury mice model

2019 ◽  
Vol 400 (6) ◽  
pp. 753-763 ◽  
Author(s):  
Xingping Dai ◽  
Min Yi ◽  
Dongsheng Wang ◽  
Yanyi Chen ◽  
Xia Xu
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Neuronal Apoptosis ◽  
Apoptotic Cell ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Luciferase Reporter ◽  
Cell Detection ◽  
Neuroprotective Effects ◽  
Newborn Mice

Abstract The present study was designed to investigate the mechanism of the traditional Chinese medicine Changqin NO. 1 on the amelioration of traumatic brain injury (TBI). Adult male C57BL/6J mice and newborn mice were used to generate a mouse TBI model and harvest primary neurons, respectively. The localizations of specific neural markers neuropilin-1 (Nrp-1), growth-associated protein-43 (GAP-43) and microtubule-associated protein Tau (Tau) were examined in brain tissues by immunohistochemistry. Terminal deoxynucleotidyl transferase dUTP nick end labeling apoptotic cell detection in tissue sections and the CCK-8 cell viability assay were performed to examine neuronal apoptosis. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were also carried out in this study. The association between long non-coding RNA (lncRNA) growth-arrest specific 5 (GAS5), miR-335 and RAS p21 GTPase activating protein 1 (Rasa1) was disclosed using the dual-luciferase reporter assay. Changqin NO. 1 inhibited TBI-induced neuronal apoptosis in vivo and in vitro. GAS5 functioned as a competing endogenous RNA (ceRNA) by sponging miR-335 to upregulate Rasa1 expression in mouse neuronal cells. Further investigations demonstrated that GAS5 promoted neuronal apoptosis following TBI via the miR-335/Rasa1 axis. In vivo experiments indicated that Changqin NO. 1 exerted neuroprotection during TBI via the GAS5/miR-335/Rasa1 axis. Changqin NO. 1 promoted neuroprotective effects by inhibiting neuronal apoptosis via the GAS5/miR-335/Rasa1 axis in TBI.

scholarly journals LncRNA SNHG15 contributes to doxorubicin resistance of osteosarcoma cells through targeting the miR-381-3p/GFRA1 axis

2020 ◽  
Vol 15 (1) ◽  
pp. 871-883
Author(s):  
Jinshan Zhang ◽  
Dan Rao ◽  
Haibo Ma ◽  
Defeng Kong ◽  
Xiaoming Xu ◽  
...  
Keyword(s):  
Cell Lines ◽  
Noncoding Rna ◽  
Xenograft Model ◽  
Inhibitory Effect ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Osteosarcoma Cell ◽  
Osteosarcoma Cells

AbstractBackgroundOsteosarcoma is a common primary malignant bone cancer. Long noncoding RNA small nucleolar RNA host gene 15 (SNHG15) has been reported to play an oncogenic role in many cancers. Nevertheless, the role of SNHG15 in the doxorubicin (DXR) resistance of osteosarcoma cells has not been fully addressed.MethodsCell Counting Kit-8 assay was conducted to measure the half-maximal inhibitory concentration value of DXR in osteosarcoma cells. Western blotting was carried out to examine the levels of autophagy-related proteins and GDNF family receptor alpha-1 (GFRA1). Quantitative reverse transcription-polymerase chain reaction was performed to determine the levels of SNHG15, miR-381-3p, and GFRA1. The proliferation of osteosarcoma cells was measured by MTT assay. The binding sites between miR-381-3p and SNHG15 or GFRA1 were predicted by Starbase bioinformatics software, and the interaction was confirmed by dual-luciferase reporter assay. Murine xenograft model was established to validate the function of SNHG15 in vivo.ResultsAutophagy inhibitor 3-methyladenine sensitized DXR-resistant osteosarcoma cell lines to DXR. SNHG15 was upregulated in DXR-resistant osteosarcoma tissues and cell lines. SNHG15 knockdown inhibited the proliferation, DXR resistance, and autophagy of osteosarcoma cells. MiR-381-3p was a direct target of SNHG15, and GFRA1 bound to miR-381-3p in osteosarcoma cells. SNHG15 contributed to DXR resistance through the miR-381-3p/GFRA1 axis in vitro. SNHG15 depletion contributed to the inhibitory effect of DXR on osteosarcoma tumor growth through the miR-381-3p/GFRA1 axis in vivo.ConclusionsSNHG15 enhanced the DXR resistance of osteosarcoma cells through elevating the autophagy via targeting the miR-381-3p/GFRA1 axis. Restoration of miR-381-3p expression might be an underlying therapeutic strategy to overcome the DXR resistance of osteosarcoma.

An Activatable NIR‐II Nanoprobe for In Vivo Early Real‐Time Diagnosis of Traumatic Brain Injury

2019 ◽  
Vol 59 (1) ◽  
pp. 247-252 ◽  
Author(s):  
Chunyan Li ◽  
Wanfei Li ◽  
Huanhuan Liu ◽  
Yejun Zhang ◽  
Guangcun Chen ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Real Time ◽  
Time Diagnosis
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