Atorvastatin ameliorates early brain injury through inhibition of apoptosis and ER stress in a rat model of subarachnoid hemorrhage

Aneurysmal subarachnoid hemorrhage (SAH) is a severe cerebrovascular disease with very poor prognosis. The aim of the present study was to evaluate the protective effects of atorvastatin on early brain injury (EBI) after SAH using a perforation SAH model. Male Sprague–Dawley rats were randomly divided into four groups: the sham group, the SAH group (model group), SAH + 10 mg.kg−1.day−1 atorvastatin (low atorvastatin group), and SAH + 20 mg.kg−1.day−1 atorvastatin (high atorvastatin group). Atorvastatin was administered orally by gastric gavage for 15 days before operation. At 24 h after SAH, we evaluated the effects of atorvastatin on brain water content, apoptosis by TUNEL assay and scanning electron microscope (SEM), and the expression of apoptosis-related proteins by immunofluorescence and Western blotting analysis. Compared with the sham group, we observed increased brain water content, significant apoptosis, and elevated levels of apoptosis-related proteins including caspase-3, CCAAT enhancer-binding protein homologous protein (CHOP), the 78-kDa glucose-regulated protein (GRP78), and aquaporin-4 (AQP4) in the SAH group. Atorvastatin administration under all doses could significantly reduce brain water content, apoptosis, and the expression levels of caspase-3, CHOP, GRP78, and AQP4 at 24 h after SAH. Our data show that early treatment with atorvastatin effectively ameliorates EBI after SAH through anti-apoptotic effects and the effects might be associated inhibition of caspase-3 and endoplasmic reticulum (ER) stress related proteins CHOP and GRP78.

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A20 Establishes Negative Feedback With TRAF6/NF-κB and Attenuates Early Brain Injury After Experimental Subarachnoid Hemorrhage

Frontiers in Immunology ◽

10.3389/fimmu.2021.623256 ◽

2021 ◽

Vol 12 ◽

Author(s):

Hong-Ji Deng ◽

QuZhen Deji ◽

WangDui Zhaba ◽

Jia-Qiang Liu ◽

Sheng-Qing Gao ◽

...

Keyword(s):

Subarachnoid Hemorrhage ◽

Brain Injury ◽

Negative Feedback ◽

Inflammatory Cytokine ◽

Neuroprotective Effect ◽

Early Brain Injury ◽

Brain Water Content ◽

Nissl Staining ◽

Cytokine Levels ◽

Brain Water

Nuclear factor (NF)-κB–ty -50mediated neuroinflammation plays a crucial role in early brain injury (EBI) after subarachnoid hemorrhage (SAH). As an important negative feedback regulator of NF-κB, A20 is essential for inflammatory homeostasis. Herein, we tested the hypothesis that A20 attenuates EBI by establishing NF-κB–associated negative feedback after experimental SAH. In vivo and in vitro models of SAH were established. TPCA-1 and lentivirus were used for NF-κB inhibition and A20 silencing/overexpression, respectively. Cellular localization of A20 in the brain was determined via immunofluorescence. Western blotting and enzyme-linked immunosorbent assays were applied to observe the expression of members of the A20/tumor necrosis factor receptor-associated factor 6 (TRAF6)/NF-κB pathway and inflammatory cytokines (IL-6, IL-1β, TNF-α). Evans blue staining, TUNEL staining, Nissl staining, brain water content, and modified Garcia score were performed to evaluate the neuroprotective effect of A20. A20 expression by astrocytes, microglia, and neurons was increased at 24 h after SAH. A20 and inflammatory cytokine levels were decreased while TRAF6 expression was elevated after NF-κB inhibition. TRAF6, NF-κB, and inflammatory cytokine levels were increased after A20 silencing but suppressed with A20 overexpression. Also, Bcl-2, Bax, MMP-9, ZO-1 protein levels; Evans blue, TUNEL, and Nissl staining; brain water content; and modified Garcia score showed that A20 exerted a neuroprotective effect after SAH. A20 expression was regulated by NF-κB. In turn, increased A20 expression inhibited TRAF6 and NF-κB to reduce the subsequent inflammatory response. Our data also suggest that negative feedback regulation mechanism of the A20/TRAF6/NF-κB pathway and the neuroprotective role of A20 to attenuate EBI after SAH.

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Ulinastatin alleviates traumatic brain injury by reducing endothelin-1

Translational Neuroscience ◽

10.1515/tnsci-2021-0001 ◽

2020 ◽

Vol 12 (1) ◽

pp. 001-008

Author(s):

Ting Liu ◽

Xing-Zhi Liao ◽

Mai-Tao Zhou

Keyword(s):

Traumatic Brain Injury ◽

Brain Injury ◽

Western Blot ◽

Water Content ◽

Brain Edema ◽

Rat Model ◽

Neurosurgical Procedures ◽

Brain Water Content ◽

The Brain ◽

Brain Water

Abstract Background Brain edema is one of the major causes of fatality and disability associated with injury and neurosurgical procedures. The goal of this study was to evaluate the effect of ulinastatin (UTI), a protease inhibitor, on astrocytes in a rat model of traumatic brain injury (TBI). Methodology A rat model of TBI was established. Animals were randomly divided into 2 groups – one group was treated with normal saline and the second group was treated with UTI (50,000 U/kg). The brain water content and permeability of the blood–brain barrier were assessed in the two groups along with a sham group (no TBI). Expression of the glial fibrillary acidic protein, endthelin-1 (ET-1), vascular endothelial growth factor (VEGF), and matrix metalloproteinase 9 (MMP-9) were measured by immunohistochemistry and western blot. Effect of UTI on ERK and PI3K/AKT signaling pathways was measured by western blot. Results UTI significantly decreased the brain water content and extravasation of the Evans blue dye. This attenuation was associated with decreased activation of the astrocytes and ET-1. UTI treatment decreased ERK and Akt activation and inhibited the expression of pro-inflammatory VEGF and MMP-9. Conclusion UTI can alleviate brain edema resulting from TBI by inhibiting astrocyte activation and ET-1 production.

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FGF10 Attenuates Experimental Traumatic Brain Injury through TLR4/MyD88/NF-κB Pathway

Cells Tissues Organs ◽

10.1159/000511381 ◽

2021 ◽

pp. 1-9

Author(s):

Qinhan Hou ◽

Hongmou Chen ◽

Quan Liu ◽

Xianlei Yan

Keyword(s):

Traumatic Brain Injury ◽

Brain Injury ◽

Protein Expression ◽

Water Content ◽

Neurological Deficit ◽

Neuronal Apoptosis ◽

Neuronal Damage ◽

Intraventricular Injection ◽

Brain Water Content ◽

Brain Water

Traumatic brain injury (TBI) can induce neuronal apoptosis and neuroinflammation, resulting in substantial neuronal damage and behavioral disorders. Fibroblast growth factors (FGFs) have been shown to be critical mediators in tissue repair. However, the role of FGF10 in experimental TBI remains unknown. In this study, mice with TBI were established via weight-loss model and validated by increase of modified neurological severity scores (mNSS) and brain water content. Secondly, FGF10 levels were elevated in mice after TBI, whereas intraventricular injection of Ad-FGF10 decreased mNSS score and brain water content, indicating the remittance of neurological deficit and cerebral edema in TBI mice. In addition, neuronal damage could also be ameliorated by stereotactic injection of Ad-FGF10. Overexpression of FGF10 increased protein expression of Bcl-2, while it decreased Bax and cleaved caspase-3/PARP, and improved neuronal apoptosis in TBI mice. In addition, Ad-FGF10 relieved neuroinflammation induced by TBI and significantly reduced the level of interleukin 1β/6, tumor necrosis factor α, and monocyte chemoattractant protein-1. Moreover, Ad-FGF10 injection decreased the protein expression level of Toll-like receptor 4 (TLR4), MyD88, and phosphorylation of NF-κB (p-NF-κB), suggesting the inactivation of the TLR4/MyD88/NF-κB pathway. In conclusion, overexpression of FGF10 could ameliorate neurological deficit, neuronal apoptosis, and neuroinflammation through inhibition of the TLR4/MyD88/NF-κB pathway, providing a potential therapeutic strategy for brain injury in the future.

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Extracellular Matrix Metalloproteinase Inducer is Associated with Severity of Brain Oedema following Experimental Subarachnoid Haemorrhage in Rats

Journal of International Medical Research ◽

10.1177/147323001204000328 ◽

2012 ◽

Vol 40 (3) ◽

pp. 1089-1098 ◽

Cited By ~ 4

Author(s):

Y Tu ◽

J Fu ◽

J Wang ◽

G Fu ◽

L Wang ◽

...

Keyword(s):

Extracellular Matrix ◽

Subarachnoid Haemorrhage ◽

Water Content ◽

Brain Oedema ◽

Sham Group ◽

Brain Water Content ◽

Extracellular Matrix Metalloproteinase Inducer ◽

Protein Levels ◽

Inhibition Group ◽

Brain Water

OBJECTIVE: Brain oedema is a major cause of clinical deterioration and death following brain trauma; cellular mechanisms involved in its development remain elusive. This study investigated the role of extracellular matrix metalloproteinase inducer (EMMPRIN) in brain oedema. METHODS: The monofilament puncture model was used to induce subarachnoid haemorrhage. Adult male Sprague—Dawley rats were divided into five groups ( n = 20 per group): sham-operated, sacrificed immediately after surgery (sham group); sacrificed 12, 24 or 72 h after subarachnoid haemorrhage induction (SAH-12, SAH-24 and SAH-72 groups, respectively); treated with EMMPRIN inhibitor immediately after subarachnoid haemorrhage, sacrificed at 24 h (SAH-inhibition group). Mean brain water content, and EMMPRIN mRNA and protein levels, were determined. RESULTS: Compared with the sham group, mean brain water content, EMMPRIN mRNA and protein levels in the SAH-12, SAH-24 and SAH-72 groups increased rapidly and significantly (maximal at 24 h). EMMPRIN inhibition significantly reduced mean brain water content and EMMPRIN mRNA and protein levels in the SAH-inhibition group, compared with the SAH-24 group. CONCLUSIONS: EMMPRIN upregulation may be important in the formation of brain oedema; inhibition of EMMPRIN activity may provide a potential approach to reduce oedema after subarachnoid haemorrhage.

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Metabolic Insight Into the Neuroprotective Effect of Tao-He-Cheng-Qi (THCQ) Decoction on ICH Rats Using Untargeted Metabolomics

Frontiers in Pharmacology ◽

10.3389/fphar.2021.636457 ◽

2021 ◽

Vol 12 ◽

Author(s):

Rui-Pei Yang ◽

Da-Ke Cai ◽

Yu-Xing Chen ◽

Hai-Ning Gang ◽

Mei Wei ◽

...

Keyword(s):

Lipid Metabolism ◽

Amino Acid ◽

Intracerebral Hemorrhage ◽

Water Content ◽

Sham Group ◽

Neuroprotective Effect ◽

Treated Group ◽

Brain Water Content ◽

Model Group ◽

Brain Water

Tao-He-Cheng-Qi decoction (THCQ) is an effective traditional Chinese medicine used to treat intracerebral hemorrhage (ICH). This study was performed to investigate the possible neuroprotective effect of THCQ decoction on secondary brain damage in rats with intracerebral hemorrhage and to elucidate the potential mechanism based on a metabolomics approach. Sprague-Dawley (SD) rats were randomly divided into five groups: the sham group, collagenase-induced ICH model group, THCQ low-dose (THCQ-L)-treated group, THCQ moderate-dose (THCQ-M)-treated group and THCQ high-dose (THCQ-H)-treated group. Following 3 days of treatment, behavioral changes and histopathological lesions in the brain were estimated. Untargeted metabolomics analysis with multivariate statistics was performed by using ultrahigh-performance liquid chromatography–mass spectrometry (UPLC-Q-Exactive Orbitrap MS). THCQ treatment at two dosages (5.64 and 11.27 g/kg·d) remarkably improved behavior (p < 0.05), brain water content (BMC) and hemorheology (p < 0.05) and improved brain nerve tissue pathology and inflammatory infiltration in ICH rats. Moreover, a metabolomic analysis demonstrated that the serum metabolic profiles of ICH patients were significantly different between the sham group and the ICH-induced model group. Twenty-seven biomarkers were identified that potentially predict the clinical benefits of THCQ decoction. Of these, 4 biomarkers were found to be THCQ-H group-specific, while others were shared between two clusters. These metabolites are mainly involved in amino acid metabolism and glutamate-mediated cell excitotoxicity, lipid metabolism-mediated oxidative stress, and mitochondrial dysfunction caused by energy metabolism disorders. In addition, a correlation analysis showed that the behavioral scores, brain water content and hemorheology were correlated with levels of serum metabolites derived from amino acid and lipid metabolism. In conclusion, the results indicate that THCQ decoction significantly attenuates ICH-induced secondary brain injury, which could be mediated by improving metabolic disorders in cerebral hemorrhage rats.

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Abstract 2611: Inhibition Of Rho-kinase By Hydroxyfasudil Attenuates Early Brain Injury After Subarachnoid Hemorrhage In Rats

Stroke ◽

10.1161/str.43.suppl_1.a2611 ◽

2012 ◽

Vol 43 (suppl_1) ◽

Author(s):

Mutsumi Fujii ◽

Kamil Duris ◽

Orhan Altay ◽

Yoshiteru Soejima ◽

Prativa Sherchan ◽

...

Keyword(s):

Brain Injury ◽

Brain Edema ◽

Rho Kinase ◽

Early Brain Injury ◽

Vehicle Group ◽

Brain Water Content ◽

Single Treatment ◽

Ipsilateral Hemisphere ◽

Neurological Score ◽

Brain Water

Background: Early brain injury comprising of blood-brain barrier (BBB) disruption and brain edema is an important part of subarachnoid hemorrhage (SAH) pathophysiology. Although several reports have shown that inhibition of Rho-kinase (ROCK) plays an important role in suppressing barrier permeability, the effect of ROCK on the BBB in SAH is not well understood. Therefore, we assessed the hypothesis that (A) ROCK inhibitors, hydroxyfasudil (HF) and Y27632 attenuate early brain injury after SAH, and (B) HF decreases brain edema via preservation of the BBB. Methods: Adult male rats were randomly assigned to five groups; sham-operated, SAH with saline, SAH with HF (10mg/kg, i.p.) treatment at 0.5 hrs, SAH with HF (10mg/kg, i.p.) treatment at 0.5 and 6 hrs each , and SAH with Y27632 (10mg/kg, i.p.) treatment at 0.5 hrs. The endovascular perforation method was used to produce SAH. Neurological scores were evaluated before sacrifice at 24 and 72 hours after injury. Brain water content, Evans blue dye extravasation assay, Rho-kinase activity assay, and Western blotting analyses were performed. Results: Among 175 surgeries performed, 53 animals were excluded. Of those excluded, 26 died from severe SAH and 27 had only mild SAH. There were no differences between the SAH with saline group (Vehicle group), SAH with HF group, and SAH with Y27632 group in SAH grading and mortality. HF but not Y27632 significantly improved neurological outcomes. Both groups significantly attenuated brain water content (BWC) in the ipsilateral hemisphere compared with the Vehicle group at 24 hrs after SAH for single treatment. The BWC of the HF group was also significantly lower than that of the Vehicle group in the contralateral hemisphere and cerebellum. There was no difference in neurological score between the Vehicle and HF group with two treatments, however, HF significantly ameliorated BWC in the ipsilateral hemisphere compared with vehicle at 24 hrs after SAH. There was no difference in neurological score and BWC between the Vehicle and HF groups with single treatment at 72 hrs after SAH. Evans blue extravasation in the ipsilateral hemisphere in the Vehicle group was significantly higher than in the HF group for single treatment at 24 hours after SAH. Supernatants of the ipsilateral hemisphere of single treatment groups sacrificed at 24 hours showed significantly higher ROCK activity in the Vehicle group than the sham group, and the HF group showed significantly lower activity than the Vehicle group. As the tight junction (TJ) is one of the components of the endothelial junctional complex in the BBB, we evaluated TJ proteins, occludin and Zona occludens-1 (ZO-1) in Western blotting analyses. Occludin and ZO-1 levels were significantly lower in the Vehicle group than the HF group. Conclusion: The ROCK inhibitor, HF, attenuates early brain injury by decreasing brain edema after SAH via protection of tight junction proteins.

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Tauroursodeoxycholic acid attenuates neuronal apoptosis via the TGR5/ SIRT3 pathway after subarachnoid hemorrhage in rats

Biological Research ◽

10.1186/s40659-020-00323-1 ◽

2020 ◽

Vol 53 (1) ◽

Author(s):

Huihui Wu ◽

Nini Yu ◽

Xia Wang ◽

Yina Yang ◽

Hui Liang

Keyword(s):

Subarachnoid Hemorrhage ◽

Water Content ◽

Neuronal Apoptosis ◽

Critical Event ◽

Tunel Staining ◽

Protective Effects ◽

Brain Water Content ◽

Sprague Dawley ◽

Tauroursodeoxycholic Acid ◽

Brain Water

Abstract Background Neuronal apoptosis plays a critical event in the pathogenesis of early brain injury after subarachnoid hemorrhage (SAH). This study investigated the roles of Tauroursodeoxycholic acid (TUDCA) in attenuate neuronal apoptosis and underlying mechanisms after SAH. Methods Sprague–Dawley rats were subjected to model of SAH and TUDCA was administered via the internal carotid injection. Small interfering RNA (siRNA) for TGR5 were administered through intracerebroventricular injection 48 h before SAH. Neurological scores, brain water content, Western blot, TUNEL staining and immunofluorescence staining were evaluated. Results TUDCA alleviated brain water content and improved neurological scores at 24 h and 72 h after SAH. TUDCA administration prevented the reduction of SIRT3 and BCL-2 expressions, as well as the increase of BAX and cleaved caspase-3.Endogenous TGR5 expression were upregulated after SAH and treatment with TGR5 siRNA exacerbated neurological outcomes after SAH and the protective effects of TUDCA at 24 h after SAH were also abolished by TGR5 siRNA. Conclusions Our findings demonstrate that TUDCA could attenuated neuronal apoptosis and improve neurological functions through TGR5/ SIRT3 signaling pathway after SAH. TUDCA may be an attractive candidate for anti-apoptosis treatment in SAH.

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Tetramethylpyrazine Protects against Early Brain Injury after Experimental Subarachnoid Hemorrhage by Affecting Mitochondrial-Dependent Caspase-3 Apoptotic Pathway

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2017/3514914 ◽

2017 ◽

Vol 2017 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Shaoxue Li ◽

Xiaolan Xiao ◽

Xiaojia Ni ◽

Zhennan Ye ◽

Junjie Zhao ◽

...

Keyword(s):

Subarachnoid Hemorrhage ◽

Brain Injury ◽

Caspase 3 ◽

B Cell Lymphoma ◽

Early Brain Injury ◽

Calcium Overload ◽

Mitochondrial Morphology ◽

Apoptotic Pathway ◽

Experimental Subarachnoid Hemorrhage ◽

Cyt C

This study was to test the hypothesis that tetramethylpyrazine (TMP) protected against early brain injury after subarachnoid hemorrhage (SAH) by affecting the mitochondrial-dependent caspase-3 apoptotic pathway. TMP was administrated after the rats’ prechiasmatic SAH mode. Animal neurobehavioral functions were assessed and the mitochondrial morphology, mitochondrial and cytoplasmic calcium, and mitochondrial membrane potential changes (Δψm) of the brain tissues were measured. The expressions of cytoplasmic cytochrome c (cyt c), second mitochondria-derived activator of caspases (Smac), and cleaved caspase-3 B-cell lymphoma 2 (bcl-2) in cells were determined and cellular apoptosis was detected. The treatment of TMP resulted in less apoptotic cells and milder mitochondrial injury and potentially performed better in the neurobehavioral outcome compared to those with saline. Also, TMP ameliorated calcium overload in mitochondria and cytoplasm and alleviated the decrease of Δψm. In addition, TMP inhibited the expression of cytoplasmic cyt c, Smac, and cleaved caspase-3, yet it upregulated the expression of bcl-2. These findings suggest that TMP exerts an antiapoptosis property in the SAH rat model and this is probably mediated by the caspase-3 apoptotic pathway triggered by mitochondrial calcium overload. The finding offers a new therapeutic candidate for early brain injury after SAH.

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Role of flunarizine hydrochloride in secondary brain injury following intracerebral hemorrhage in rats

International Journal of Immunopathology and Pharmacology ◽

10.1177/0394632017742224 ◽

2017 ◽

Vol 30 (4) ◽

pp. 413-419 ◽

Cited By ~ 4

Author(s):

Jianping Niu ◽

Rui Hu

Keyword(s):

Brain Injury ◽

Intracerebral Hemorrhage ◽

Water Content ◽

Cell Apoptosis ◽

Akt Pathway ◽

Secondary Brain Injury ◽

Brain Water Content ◽

Hematoma Volume ◽

Brain Water

This study aimed to explore the role and mechanism(s) of flunarizine hydrochloride in the intracerebral hemorrhage (ICH) rats. The 32 adult male Sprague Dawley (SD) rats were randomly assigned into four groups: control group, sham group, ICH group, and FLU + ICH group. The effects of flunarizine hydrochloride were assessed on the basis of hematoma volume, blood–brain barrier (BBB) integrity, and brain water content in the ICH rat models. The role of flunarizine hydrochloride in cell recovery was assessed by behavioral scores, quantitative real-time polymerase chain reaction (qRT-PCR), and western blot assay. Involvement of PI3K/AKT pathway in exerting the effect of flunarizine hydrochloride was also determined. Results showed that the hematoma volume, BBB integrity, and brain water content were significantly decreased in the FLU + ICH group. Cell apoptosis significantly increased in the ICH model group, while flunarizine hydrochloride decreased this increase. The expressions of glial cell line-derived neurotrophic factor (GDNF), neuroglobin (NGB), and p-AKT were increased after flunarizine hydrochloride treatment in ICH rats. In conclusion, flunarizine hydrochloride has protective effects against ICH by reducing brain injury, cell apoptosis, and the activation of P13K/AKT pathway. These findings provide a theoretical basis for the treatment of flunarizine hydrochloride in ICH.

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