NOD-like-receptor signaling pathway mediates inflammation and triggers cerebral injury in cerebral venous thrombosis

doi:10.21203/rs.3.rs-2975353/v1

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NOD-like-receptor signaling pathway mediates inflammation and triggers cerebral injury in cerebral venous thrombosis

https://doi.org/10.21203/rs.3.rs-2975353/v1

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Background

Cerebral venous thrombosis (CVT) is a special type of stroke with an increasing incidence. However, the pathophysiological mechanisms remain elusive, which hinders a comprehensive understanding of CVT. We used a CVT model in rats to elucidate the mechanism of neurological damage.

Methods

We constructed a CVT model to examine neurological function and performed neuroimaging. RNA-Seq and biological information technology were utilized to analyze the transcriptome features of the Sham, middle cerebral artery occlusion (MCAO), and CVT groups, subsequently selecting significantly upregulated signaling pathways in the CVT rat brain. The activation of signaling pathways and immune cells in CVT was confirmed through flow cytometry (FC), real-time quantitative polymerase chain reaction (qPCR), and immunofluorescence staining (IF).

Results

Twenty-four hours after CVT establishment, rats exhibited significant Magnetic resonance imaging (MRI)-T2 hyperintensity and neurological impairment compared to sham rats. Transcriptome profiling showed that the inflammatory response was a significant and specific characteristic of the CVT group compared with the sham and MCAO groups. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of differentially expressed genes (DEGs) indicated that the DEGs were mainly enriched in the gene set of inflammation-related responses. Single-sample gene set enrichment analysis (ssGSEA) also suggested that the immune inflammatory response score was increased significantly. Furthermore, Immune-AI mouse revealed that microglia were the most significantly elevated immune inflammatory cells after CVT. GSEA indicated that the nucleotide-binding oligomerization domain (NOD)-like-receptor signaling pathway was significantly upregulated compared to other inflammatory signaling pathways, and then, key driver analysis (KDA) of DEGs in the NOD-like-receptor signaling pathway revealed that Nod-2 and other genes were the core genes. Importantly, inhibiting the NOD-like-receptor signaling pathway in CVT rats resulted in neurological function improvement and infarct size reduction.

Conclusions

The microglia-mediated inflammatory response and NOD-like-receptor signaling pathway activation are significant pathological changes in brain injury after CVT. This study may enhance comprehension of the pathological mechanisms underlying CVT and provide novel insights for further investigation into injuries in CVT.

stroke

cerebral venous thrombosis

inflammation

microglial

NOD-like-receptor signaling pathway

CVT is a particular stroke subtype, also known as venous stroke, which is characterized by infarction with increased intracranial pressure and focal neurologic deficits and mainly occurs in young and middle-aged individuals[1–3]. As a neurovascular emergency, in contrast to arterial stroke, CVT remains uncertain, resulting in a delayed or missed diagnosis, with an average delay of 7 days and a missed diagnosis rate of up to 73%[4–7]. A few studies have reported that hypoxia-inducible factor 1-alpha (HIF-1α)[8], inflammation[9], cerebral edema[10] and blood‒brain barrier breakdown [11] are associated with CVT, yet much remains to be learned about its underlying pathophysiology. Neutrophils/lymphocytes (NLR)[12] and inflammatory cytokines such as C-reactive protein (CRP)[13] and interleukin-6 (IL-6)[14] in peripheral blood were significantly elevated, which was associated with severity and long-term prognosis after CVT. However, the molecular and pathological features, as well as the mechanisms in CVT brain tissue, have rarely been studied.

In the present study, we utilized a CVT rat model to investigate the injury mechanism of cerebral infarction. We found that the inflammatory response is represented in the transcriptome of brain injury after CVT, and the NOD-like-receptor signaling pathway was upregulated significantly in CVT injury locations, especially in microglia. These findings are consistent with the clinical finding of the significant elevation of inflammatory markers in the peripheral blood after CVT.

Establishment of animal models

A total of 6- to 8-week-old female wild-type rats were purchased and used at the Department of Laboratory Animal Science of Capital Medical University, and kept under standard conditions in our local animal facility (Laboratory Animal Management Center, Capital Medical University) in pathogen-free cages with food and water available ad libitum. All studies were conducted in accordance with the Institutional Guide for the Care and Use of Laboratory Animals and were approved by Capital Medical University Institutional Animal Care and Use Committee (Protocol Nos. AEEI-2018-121).

The rats were anesthetized with 5% isoflurane and maintained intraoperatively with a mixture of 1–2% enflurane, delivered through a face mask in combination with 70% nitrous oxide and 30% oxygen. The animals were positioned in a prone posture and their heads were immobilized at the midline using a stereotaxic frame. Throughout the experiment, a thermostatically controlled heating pad was utilized to maintain the temperature at 37 ± 0.5°C. First, a 1.5-cm incision was made at the midline of each rat's head, followed by dissection of subcutaneous tissue to expose the skull. A longitudinal cranial window (10 × 4 mm) was created between the bregma and lambda, exposing the superior sagittal sinus and bilateral cortex through microscopic observation using a high-speed dental drill. During the drilling process, the drill tip was continuously irrigated with normal saline to prevent thermal damage to the dura mater and cortex. Subsequently, within 3 minutes, a microinjector was used to inject thrombin into the sinus cavity of the segment. After injection, the injection needle was kept in place for 30s before retraction to avoid reflux.

The surgical site was lavaged with normal saline, followed by occlusion of the cranial aperture using bone wax in each cohort. Postoperatively, ketoprofen was subcutaneously administered to each rat for analgesia. Sham operation consisted of bilateral surgical exposure of both the internal jugular veins and posterior glenoid vein without surgical ligation and exposed the superior sagittal sinus (SSS) and without inject thrombin, with the same length of anesthesia.

Application of the Pericam Perfusion Speckle Imager system

The Pericam Perfusion Speckle Imager (PSI) employs a 785 nm invisible laser to measure blood perfusion in the cerebral ischemic model pre- and post-modeling. We designated the SSS as the region of interest and examined changes in cerebral blood flow during surgery to confirm successful induction of thrombosis and validate the CVT rat model.

MRI experiments

MRI examinations were performed using a 7.0-T MRI scanner (BRUKER PharmaScan, Germany). Among them, T2-weighted imaging was utilized for the assessment of infarct volume. The rats were anesthetized with isoflurane. Respiration and cardiac activity were continuously monitored during scanning. T2-weighted imaging was acquired with the following parameters: matrix = 256 × 256, field of view = 3.3 cm × 3.3 cm, repetition time = 3600 ms, echo time = 12 ms, slice thickness = 0.8 mm. The infarcts were confirmed by high signals acquired from T2-WI. The infarct volume was calculated as infarcted volume/ total volume × 100% according to T2-WI.

Rotarod test

Motor coordination was evaluated using an automated three-lane rotarod. The rats were placed on the rotarod to acclimate for 10 seconds at an initial speed of 5 rpm/min, followed by acceleration from 5 rpm/min to 40 rpm/min within five minutes. Rats underwent a five-day training period to ensure their ability to run on the road proficiently. The duration and speed to fall were recorded.

Novel object recognition tests

48 hours after being exposed to CVT, rats were subjected to a novel object recognition test. Two identical objects were positioned at opposite ends of the box during the training session, while a rat was placed in the center to explore for 5 minutes. Any subject that spent less than 20 seconds exploring both objects was excluded from further analysis. 24 hours later, a novel object and a familiar object were reintroduced into the same locations as in the training phase. The rat was placed in the center of the arena and allowed to freely explore for 5 minutes while being monitored by Smart 3.0 behavioral tracking software. Animal behavior was recorded by Smart 3.0 behavioral tracking software. The ratio of time spent with the novel object to the total exploring time was calculated.

RNA-sequencing and DEGs analysis

The RNA sequencing was conducted according to the BGI-tech pipeline. Total RNA underwent mRNA enrichment: Magnetic beads with OligodT were utilized to enrich the polyA-tailed mRNA. The resulting RNA was segmented using an interrupting buffer, and then subjected to reverse transcription with random N6 primers. Finally, double-stranded DNA was synthesized from the resulting cDNA two-strand. The synthetic double-stranded DNA is flattened and phosphorylated at the 5’-end, with a sticky end protruding an “A” at the 3′-end and a bubbling-like connector protruding “T” also at the 3′-end. The linked products were subsequently amplified by PCR using specific primers. Primary sequencing data were produced by MGISEQ-2000. Following quality control, the raw reads underwent filtration to obtain clean reads and were subsequently subjected to analysis using the tophat and cufflinks algorithms. GSEA was performed in R using the ClusterProfiler package with specific parameters: nPerm = 1000, minGSSize = 10, maxGSSize = 500, P-valueCutoff = 0.05, and pAdjustMethod = “BH”.

FC and cell sorting

Following the extraction of brain tissue from both the infarct and corresponding site, the tissues were initially subjected to a 30-minute digestion at 37°C using collagenase before being passed through a 70µM cell strainer. Single cell suspensions were then used for flow sorting or analysis by flow cytometry. For cell sorting experiments, single-cell suspensions were incubated 30min with appropriate antibodies: CD45-PE and CD11b/C-APC and were subsequently analyzed with a FACS Aria instrument.

RNA extraction and qPCR

Total RNA from brain tissues was extracted using RNAprep pure tissue kit. cDNA was synthesized using 5×FastKing-RT SuperMix. The specific products of target gene were amplified by quantitative PCR using PerfectStart Green qPCR SuperMix. GAPDH was used as the normalization control. Supplementary Table 1 lists the primer sequences used for RT-qPCR analysis. Data were analyzed using the 2^−∆∆CT approach.

IF staining

Tissue sections were incubated with primary antibodies: Nod-2and Iba-1overnight at 4°C. Samples were then washed in PBST and incubated with appropriate secondary antibodies for 1 h at room temperature. Nuclei were stained with DAPI for 3 minutes at room temperature. Images were acquired with a LSM880 Airyscan inverted laser confocal microscope and analyzed with ImageJ.

Statistical analysis

Statistical analyses were conducted using GraphPad Prism 8.0 and R 4.3.2, with all data presented as mean ± SEM from three to five independent experiments. Student's t-test for unpaired observations was employed where appropriate. A P-value < 0.05 was deemed statistically significant.

CVT contributed to poorer outcome

Cerebral blood flow perfusion diagrams corresponding to the CVT and Sham groups assisted by the laser speckle meter are shown in Fig. 1A. Compared with the Sham group, the SSS blood flow signal disappeared in the CVT group, indicating complete interruption of SSS cerebral perfusion (cerebral blood flow was 713.20 ± 58.87 PU and 258.40 ± 47.72 PU for the sham and CVT groups, respectively) (Fig. 1B). Based on the successful establishment of the CVT rat model, we detected the location and volume of ischemic and/or hemorrhagic foci in rats after CVT by MRI. The formation of ischemic injury in the brains of rats 48 hours after CVT is shown in Fig. 1C. Rats in the CVT group had a focal lesion lying in regions on both sides of the SSS and spread to the adjacent cortex, with a maximum infarction area of 10.86 ± 1.66% of the whole brain (Fig. 1D). In contrast, no imaging manifestation changes in the Sham group rats were observed. Furthermore, the results of neurological symptom score (NSS) revealed that compared to the sham group, neurological function was significantly impaired in CVT rats 48 h postoperatively (NSS was 0.40 ± 0.49 and 2.2 ± 0.40 for the Sham and CVT groups, respectively) (Fig. 1E). In the novel object recognition test assessing recognition memory in rats, the CVT group spent less time exploring novel objects than the sham group, which suggests that CVT resulted in cognitive impairment (Fig. 1F). The rotarod test showed that rats in the CVT group spent 141.88 ± 16.03 s less on the rod than those in the sham group (294.7 ± 10.60 s) (Fig. 1G). Furthermore, the velocity was correspondingly diminished (Fig. 1H). Together, these results demonstrated that CVT contributed to cerebral infarction in rats with neurological deficits.

The NOD-like-receptor signaling pathway is a key inflammatory response signaling pathway of CVT

To further explore the specific pathological mechanism of brain damage after CVT, tissue was isolated from the Sham, MCAO and CVT groups of rats for RNA sequencing with three biological repeats (Supplement Fig. 1). RNA-seq results showed that there were 5266 genes with altered expression in MCAO vs. Sham groups and no alteration in CVT vs. Sham groups (Fig. 2A). Additionally, 208 DEGs showed changed expression in CVT vs. Sham groups (Fig. 2A). There were 1446 common DEGs in the MCAO vs. sham and CVT vs. sham groups (Fig. 2A). The RNA-Seq heatmap accurately represents the significant DEGs, showing good intragroup consistency and intergroup variability (Fig. 2B). DEGs were further mapped to the databases for functional annotation and enrichment analysis. KEGG enrichment analysis indicated that inflammation-related signaling pathways such as the Nod-like-receptor signaling pathway, complement and coagulation cascade, tumor necrosis factor (TNF) and nuclear factor-k-gene binding (NF-KB) signaling pathway were associated with DEGs specific to CVT vs. Sham (P ≤ 0.05) (Fig. 2C). Then, ssGSEA was used to obtain the inflammatory enrichment score, indicating that the genes in the inflammatory response were upregulated in the CVT group (Fig. 2D). Using ImmuneAI -Mouse, a cell type proportion analysis was performed on Bulk RNA-seq brain gene expression data. The major cell type-specific genes of brain tissues were deconvolved, which indicated that the relative abundance of microglia was higher in the CVT group (Fig. 2E-F). The results of GSEA demonstrated that the TNF signaling pathway (NES:2.44, FDR:0), NOD-like-receptor signaling pathway (NES:2.45, FDR:0), NF-kB signaling pathway (NES:2.21, FDR:0) and complement and coagulation cascade pathway (NES:2.50, FDR:0) were significantly enriched in CVT brain tissue (Fig. 2G-J), which indicates that acute inflammatory response may couple with the course of CVT. Importantly, the Nod-like-receptor signaling pathway is upstream of inflammatory signaling pathways such as TNF, NF-kB, and other inflammatory factors, which mediate acute inflammatory response damage. Furthermore, KDA of 167 differentially expressed genes in the NOD-like-receptor signaling pathway revealed that "LOC100910771, Casp1, Nfkbia, Fadd, Nlrp-3, Nod-2, IL-18, Irf-3," Irf-7, Tbk-1, Ripk-1, Tab-2, Ikbkg, Chuk, Mavs, Traf-2, Tab-1, Tab-3, Ripk-2, Traf-3, Ikbke, Ikbkb, Nod-1, Mapk9, Mefv, Xiap, Casp8, Tnfaip-3, Traf-5, Nfkb1” were the core genes, and the average expression levels of these genes were significantly upregulated in the CVT group (Fig. 2K-L).

Activation of Nod-like-receptor signaling pathways and microglia in the rat brain after CVT

To validate the RNA-seq results, we selected 24 pivotal driver genes (Casp1, Nfkbia, Fadd, Nlrp-3, Nod-2, IL-18, Irf-3, Irf-7, Tbk-1, Ripk-1, Tab − 2, Ikbkg, Traf − 2, Tab − 1, Ripk − 2, Traf − 3, Ikbke, Ikbkb, Nod − 1, Mapk − 9, Mefv, Xiap, Casp8, Nfkb1) for real-time quantitative PCR validation. The results demonstrated that the expression of these key driver genes, especially Nod-2, was significantly increased in the infarct area of CVT rats, which exhibited a consistent trend with the RNA-seq data (Fig. 3A). These findings suggest that microglia play an essential role in the inflammatory response after CVT. Next, we attempted to determine the effects of CVT on microglia via flow cytometry. We discovered that compared to the Sham group, microglial (CD45⁺CD11b/c⁺) infiltration increased significantly in the infarction focus of CVT rats (Fig. 3B-C). To further confirm the role of the NOD-like-receptor signaling pathway, CD45⁺ CD11b/c⁺ microglial cells were sorted from the CVT and Sham groups by flow cytometry, and the expression of key driver genes was analyzed by qPCR. The findings indicated that the CVT group exhibited upregulated expression of the core genes involved in the NOD-like-receptor signaling pathway (Fig. 3D). Immunofluorescent double staining showed a marked increase in Iba-1⁺ microglial infiltration around the infarction area and a significant increase in Nod-2 expression in Iba-1⁺ microglia (Fig. 3E-G). Taken together, these results suggest that microglial cells are actively involved in brain injury after CVT and that the NOD-like-receptor signaling pathway is the pivotal signaling pathway.

Inhibition of the NOD-like-receptor signaling pathway ameliorated the outcome of CVT and reduced microglial activation

To verify that the NOD-like-receptor signaling pathway is involved in the inflammatory response of CVT and triggers cerebral injury, we utilized GSK717[15], a potent and selective inhibitor of Nod-2, to block the NOD-like-receptor signaling pathway. To investigate whether blocking the NOD-like-receptor signaling pathway could improve neurological deficits after CVT, we performed MRI and behavioral evaluation. T2-weighted imaging showed that the maximum infarction areas were reduced by approximately 6.18 ± 0.62% following GSK717 administration (Fig. 4A-B). Moreover, the injection of GSK717 resulted in an improvement in nervous system function after CVT (NSS scores were 0.20 ± 0.40, 2.20 ± 0.40 and 1.20 ± 0.40 for the Sham, CVT + Ctrl and CVT + GSK717 groups, respectively) (Fig. 4C). Rats in the CVT + GSK717 group spent more time investigating the new object compared with the CVT group (discrimination index is 65.7 ± 4.39%, 42.75 ± 2.93% and 61.07 ± 2.89% for Sham, CVT + Ctrl and CVT + GSK717 groups, respectively) (Fig. 4D). In parallel, the results of the rotarod test showed that GSK717 enhanced the rotarod test time (292.62 ± 4.46 s, 125.3 ± 7.56 s and 221 ± 7.77 s for the Sham, CVT + Ctrl and CVT + GSK717 groups, respectively) and speed (38 ± 1.1 r/min, 18.6 ± 1.0 r/min and 29.4 ± 1.0 r/min for the Sham, CVT + Ctrl and CVT + GSK717 groups, respectively) for rats at 48 h after CVT (Fig. 4E-F). Therefore, blocking the Nod-like-receptor signaling pathway improved the symptoms of neurological impairment after CVT by suppressing the inflammatory response.

Inhibition of Nod-2 with GSK717 resulted in blunted signal transduction in the NOD-like-receptor pathway. qPCR analysis confirmed the downregulation of expression in the infarct area of the CVT + GSK717 group (Fig. 4G). To further confirm whether GSK717 attenuated the CVT-induced inflammatory response by suppressing the NOD-like-receptor pathways, CD45⁺ CD11b/c⁺ microglial cells were quantified and sorted by flow cytometry (Fig. 4H). The results showed that microglial numbers were significantly decreased in the infarction location in the CVT + GSK717 group compared with the CVT group (Fig. 4I). Similarly, the expression of core driver genes was also reduced significantly in microglial cells (Fig. 4J). Moreover, immunofluorescence staining also revealed that blocking the NOD-like-receptor signaling pathway by GSK717 reduced Iba-1⁺ microglial cell numbers and the expression of Nod-2 in the infarct area (Fig. 4K-M). These findings suggest that microglial cells are essential for the activation of inflammatory reactions after CVT and that the NOD-like-receptor signaling-mediated inflammatory response may play an important trigger role in cerebral injury.

Stroke is a prominent contributor to both disability and mortality worldwide and can be classified into arterial stroke and venous stroke[16]. Nonetheless, in contrast to arterial stroke, venous stroke has a dearth of research regarding its pathological mechanisms. This study utilized rat animal models of MCAO and CVT to investigate the pathological mechanisms underlying arterial stroke and CVT, indicating an increase in the NOD-like-receptor signaling pathway and microglial cells in CVT infarct tissue, suggesting that the inflammatory response plays a role in brain injury after CVT. Our research provides innovative insights into the underlying pathological mechanism of CVT.

CVT involves both infarction and hemorrhage, and its pathological mechanism is intricate[17]. Due to the limited availability of brain tissue samples, clinical studies can only investigate molecular changes in peripheral blood and cerebrospinal fluid samples from patients. Our team confirmed a significant correlation between the severity of CVT and adverse outcomes with peripheral blood inflammatory markers, including IL-6, CRP, NLR and systemic immune–inflammation index[14, 18]. In addition, Aguiar et al. demonstrated that elevated levels of IL-6, CRP, and NLR may serve as prognostic markers for poorer outcomes in patients with CVT[13]. The utilization of CVT models is essential for researching the molecular pathological changes and injury mechanisms in damaged brain tissue. Our study has further demonstrated an elevated inflammatory response in tissues affected by CVT infarction, which further verifies the association between inflammation and post-CVT injury.

In this study, ImmuneAI-Mouse detected a significant increase in microglial number, which was further confirmed by FC and IF analysis. Microglia, the immune cells that reside in the central nervous system, activate and switch their phenotype in response to changes in the process of disease[19]. Microglia are derived from monocytes in the periphery and serve a dual function in disease[20]. In the acute phase, M1 macrophages can release cytotoxic factors that have a proinflammatory effect, while M2 macrophages secrete anti-inflammatory factors, stimulate angiogenesis, remove cell debris, and promote repair of peripheral nerve injuries[21]. This study demonstrates that microglial cells in the infarcted tissue exhibit an increase in number 48 hours post CVT, indicating a pivotal role as primary effector cells responsible for the inflammatory response following CVT. The number and phenotype of microglia undergo dynamic changes during different stages of arterial stroke, which are closely associated with the severity of the injury and the prognosis[22]. The multiple roles of microglia throughout the acute, subacute, and chronic phases of CVT deserve further investigation.

Our results showed that the NOD-like-receptor signaling pathway is the most important signaling pathway for mediating cerebral injury after CVT. NOD-like-receptors play an essential role in innate immunity and inflammation[23]. Chauhan et al. recently demonstrated that microglial upregulation of Nod-2 plays a crucial role in inflammatory responses following bacterial infection of the central nervous system[24]. Nod-2 recognizes muramyl dipeptide and initiates downstream signaling pathways, including the activation of NF-KB and mitogen-activated protein kinases (MAPK)s and the expression of cytokines IL-1β and IL-6, ultimately resulting in the induction of inflammatory immune responses[25, 26]. Our data clearly demonstrated that the expression of Nod-2 was rapidly and significantly increased in microglia in CVT brain infarction tissues following insult. Transcriptome profiling showed that the NF-KB signaling pathway and NOD-like-receptor signaling pathways exhibited consistent alterations. The inhibition of the NOD-like-receptor signaling pathway in CVT rats resulted in a decrease in infarct size and an improvement in neurological function. These results indicated that the NOD-like-receptor signaling pathway may exert a pivotal role in regulating inflammatory responses during cerebral ischemia injury.

This study showed that the NOD-like-receptor signaling pathway and microglial cells mediate inflammation, which is a prominent feature of pathological injury following CVT, consistent with previous research findings. This increases our comprehension of the pathological mechanism underlying CVT and provides novel insights for further investigation.

CVT

cerebral venous thrombosis

MCAO

middle cerebral artery occlusion

flow cytometry

qPCR

real-time quantitative polymerase chain reaction

immunofluorescence staining

MRI

magnetic resonance imaging

KEGG

kyoto encyclopedia of genes and genomes

DEGs

differentially expressed genes

ssGSEA

single-sample gene set enrichment analysis

NOD

nucleotide-binding oligomerization domain

KDA

key driver analysis

HIF-1α

hypoxia-inducible factor 1-alpha

NLR

neutrophils/lymphocytes

CRP

C-reactive protein

IL-6

interleukin-6

PSI

pericam perfusion speckle imager

SSS

superior sagittal sinus

NSS

neurological symptom score

TNF

tumor necrosis factor

NF-KB

nuclear factor-k-gene binding

MAPK

mitogen-activated protein kinase

Data availability

Summary data are available from the authors on request.

Fundings

This work is supported by grants from the National Natural Science Foundation of China (82271311), National Natural Science Foundation of China (82027802), Talents Gathering Project of Xuanwu Hospital Capital Medical University and Technological Innovation and Service Capability Construction Project from Beijing Municipal Commission of Education (11000023T000002157177).

Declaration of Competing Interest

The authors declare no conflict of interest.

Ethics approval and consent to participate

All studies were conducted in accordance with the Institutional Guide for the Care and Use of Laboratory Animals and were approved by Capital Medical University Institutional Animal Care and Use Committee (Protocol Nos. AEEI-2018-121).

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

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No competing interests reported.

Supplementfigure1.pdf
The Sham, CVT, and MCAO rat models were prepared, followed by the extract of infarcted brain tissue adjacent to the sagittal sinus for RNA-seq analysis. Subsequently, bioinformatics was employed to analyze the obtained data.
Supplementtable1.docx

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NOD-like-receptor signaling pathway mediates inflammation and triggers cerebral injury in cerebral venous thrombosis

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Abstract

Background

Methods

Results

Conclusions

Figures

Introduction

Materials and Methods

Establishment of animal models

Application of the Pericam Perfusion Speckle Imager system

MRI experiments

Rotarod test

Novel object recognition tests

RNA-sequencing and DEGs analysis

FC and cell sorting

RNA extraction and qPCR

IF staining

Statistical analysis

Results

CVT contributed to poorer outcome

The NOD-like-receptor signaling pathway is a key inflammatory response signaling pathway of CVT

Activation of Nod-like-receptor signaling pathways and microglia in the rat brain after CVT

Inhibition of the NOD-like-receptor signaling pathway ameliorated the outcome of CVT and reduced microglial activation

Discussion

Abbreviations

Declarations

References

Additional Declarations

Supplementary Files

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