Inhibition of CCR2 attenuates neuroinflammation and neuronal apoptosis after subarachnoid hemorrhage through the PI3K/Akt pathway.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:36566220", "kind": "science-record", "revision": 4, "content_hash": "sha256:54dde8712913f6807071b1dfbcf08741777116d6ed5dce7f3caea30938acfefd", "created_at": "2026-11-01T19:01:07Z", "updated_at": "2026-11-01T22:50:56Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Inhibition of CCR2 attenuates neuroinflammation and neuronal apoptosis after subarachnoid hemorrhage through the PI3K/Akt pathway.", "doi": "10.1186/s12974-022-02676-8", "pmid": "36566220", "authors": ["Qi Tian", "Yujia Guo", "Shi Feng", "Chengli Liu", "Peibang He", "Jianfeng Wang", "Wenrui Han", "Chen Yang", "Zhan Zhang", "Mingchang Li"], "journal": "Journal of neuroinflammation", "year": 2022, "abstract": "BACKGROUND: Neuroinflammation and neuronal apoptosis are closely associated with a poor prognosis in patients with subarachnoid hemorrhage (SAH). We investigated the role of C-C motif chemokine receptor 2 (CCR2) in SAH.\nMETHODS: Pre-processed RNA-seq transcriptome datasets GSE167110 and GSE79416 from the Gene Expression Omnibus (GEO) database were screened for genes differentially expressed between mice with SAH and control mice, using bioinformatics analysis. The endovascular perforation model was performed to establish SAH. RS504393 (a CCR2 antagonist) and LY294002 (PI3K inhibitor) were administered to explore the mechanism of neuroinflammation after SAH. SAH grading, neurological scoring, brain water content and blood-brain barrier (BBB) permeability determination, enzyme-linked immunosorbent assay (ELISA), western blotting, and immunofluorescence were performed. An in vitro model of SAH was induced in H22 cells by hemin treatment. The protective mechanism of CCR2 inhibition was studied by adding RS504393 and LY294002. Clinical cerebrospinal fluid (CST) samples were detected by ELISA.\nRESULTS: Expression of CCR2 was upregulated in both datasets and was identified as a hub gene. CCR2 expression was significantly upregulated in the cytoplasm of neurons after SAH, both in vitro and in vivo. RS significantly reduced the brain water content and blood-brain barrier permeability, alleviated neuroinflammation, and reduced neuronal apoptosis after SAH. Additionally, the protective effects of CCR2 inhibition were abolished by LY treatment. Finally, the levels of CCR2, inflammatory factors, and apoptotic factors were elevated in the CSF of patients with SAH. CCR2 levels were associated with patient outcomes at the 6-month follow-up.\nCONCLUSION: CCR2 expression was upregulated in both in vitro and in vivo SAH models. Additionally, inhibition of CCR2, at least partly through the PI3K/AKT pathway, alleviated neuroinflammation and neuronal apoptosis in vivo and in vitro. CCR2 levels in the CSF have a moderate diagnostic value for 6-month outcome prediction in patients with SAH.", "claims": [{"id": "claim-method", "statement": "The datasets of differential RNA-seq transcriptome data in SAH were compared with those of control mouse brain tissue using the DESeq2 software package (version 1.34.0), using R software (https://www.r-project.org/) [14].", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:36566220", "locator": "sections[4] \"Bioinformatics analysis\" paragraph 2", "quote": "The datasets of differential RNA-seq transcriptome data in SAH were compared with those of control mouse brain tissue using the DESeq2 software package (version 1.34.0), using R software (https://www.r-project.org/) [14]."}], "state": "complete"}, {"id": "claim-data-access", "statement": "All data generated or analyzed during the course of this study are included in the published article and its supplementary documents.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:36566220", "locator": "sections[40] \"notes\" paragraph 1", "quote": "All data generated or analyzed during the course of this study are included in the published article and its supplementary documents."}], "state": "complete"}, {"id": "claim-target-result", "statement": "Expression of CCR2 was upregulated in both datasets and was identified as a hub gene.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:36566220", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "Expression of CCR2 was upregulated in both datasets and was identified as a hub gene."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "Expression of CCR2 was upregulated in both datasets and was identified as a hub gene.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:36566220"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. This lane did not download the dataset or read its licence.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE167110"], "outputs": ["the source data with its repository provenance"], "instruments": [], "reagents": []}, {"id": "run_analysis", "order": 3, "name": "Run the published analysis", "kind": "analysis", "state": "complete", "blocked_on": null, "description": "Apply the paper's statistical method to the resolved series and compare the result with the published target.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE167110"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": []}, "data": {"state": "partial", "assets": [{"id": "source:pmid:36566220", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": "https://doi.org/10.1186/s12974-022-02676-8", "sha256": "sha256:6013fb1e7ca65a4cb5c35d0b747de0675ad1bc06fa2b7613ad104b6daeef2fe3", "bytes": 45159, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "10.1186/s12974-022-02676-8", "version": null, "note": "Full article fetched from PMC; the checksum covers the rendered section body, not the original XML bytes.", "structure": {"pmcid": "PMC9789920", "source": "europe_pmc_fulltext", "coverage": "whole_body_without_references", "text_hash_scope": "rendered_section_body"}}, {"id": "data:geo:GSE167110", "role": "raw", "format": "geo-series", "media_type": "application/vnd.ncbi.geo-series+json", "uri": "https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE167110", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "GSE167110", "version": null, "note": "All data generated or analyzed during the course of this study are included in the published article and its supplementary documents.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "All data generated or analyzed during the course of this study are included in the published article and its supplementary documents.", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM5093690", "GSM5093691", "GSM5093692", "GSM5093693", "GSM5093694", "GSM5093695", "GSM5093696", "GSM5093697", "GSM5093698", "GSM5093699"], "sample_count": 10, "group_sizes": null, "group_sizes_basis": null, "platform": "21103", "taxon": "Mus musculus", "title": "Hippocampal transcriptome changes after subarachnoid hemorrhage in mice", "resolution_checked_at": "2026-11-01T18:46:34Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}, {"id": "data:geo:GSE79416", "role": "raw", "format": "geo-series", "media_type": "application/vnd.ncbi.geo-series+json", "uri": "https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE79416", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "GSE79416", "version": null, "note": "All data generated or analyzed during the course of this study are included in the published article and its supplementary documents.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "All data generated or analyzed during the course of this study are included in the published article and its supplementary documents.", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM2094986", "GSM2094987", "GSM2094988", "GSM2094989", "GSM2094990", "GSM2094991"], "sample_count": 6, "group_sizes": null, "group_sizes_basis": null, "platform": "17021", "taxon": "Mus musculus", "title": "Altered expression profile of long non-coding RNA and mRNA in mice cortex around bleeding areas after subarachnoid hemorrhage", "resolution_checked_at": "2026-11-01T18:46:34Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}], "gaps": ["asset:data:geo:GSE167110 not downloaded", "asset:data:geo:GSE167110 licence not read", "asset:data:geo:GSE79416 not downloaded", "asset:data:geo:GSE79416 licence not read"]}, "analysis": {"state": "partial", "methods": [{"id": "method-1", "family": "differential_expression", "specific": "deseq2", "label": "Differential expression analysis", "dsl_operations": ["normalise_counts", "differential_expression_fit", "fdr_bh"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["count_matrix_with_condition"], "evidence": {"score": 1.0, "band": "verbatim", "quote": "The datasets of differential RNA-seq transcriptome data in SAH were compared with those of control mouse brain tissue using the DESeq2 software package (version 1.34.0), using R software (https://www.r-project.org/) [14]."}}, {"id": "method-2", "family": "group_comparison", "specific": "anova", "label": "Group comparison", "dsl_operations": ["group_summary", "welch_t", "bootstrap_mean_difference", "group_comparison_anova"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["two_or_more_groups_continuous"], "evidence": {"score": 0.8, "band": "high", "quote": "analysis of variance"}}, {"id": "method-3", "family": "diagnostic_evaluation", "specific": "roc", "label": "Diagnostic test evaluation", "dsl_operations": ["confusion_counts", "roc_curve", "sensitivity_specificity", "solve_confusion_matrix", "classification_metrics", "published_precision_envelope"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["reference_standard_and_test"], "evidence": {"score": 0.8, "band": "high", "quote": "ROC curve"}}], "targets": [], "gaps": ["method.reader_differs_from_extractor"]}, "capability": {"state": "complete", "requirements": [{"capability_id": "capability:computational_analysis", "label": "Computational analysis platform", "assay_families": ["meta_analysis", "machine_learning", "survival_analysis", "statistical_regression", "pca", "molecular_docking", "network_analysis", "differential_expression", "gwas"], "availability_state": "owned", "tier_unlocked": "R1", "candidate": "Automated Assays Laravel application and simulated adapters"}], "gaps": []}, "readiness": {"state": "complete", "tier": "R1", "blocking_axis": "none", "blocked_on": "nothing:replicable_now", "axes": {"method": {"tier": "R1", "expressible": true, "required_operations": ["normalise_counts", "differential_expression_fit", "fdr_bh"], "missing_operations": [], "basis": "The paper names the method it used and the corpus's method extractor maps it onto the DSL catalogue: deseq2."}, "capability": {"tier": "R1", "capability_id": "capability:computational_analysis", "availability_state": "owned", "basis": "The computational analysis is owned. The physical bench the paper used is not, so a physical validation cannot be run here."}, "data": {"tier": "R1", "reachable": true, "accessions": ["GSE167110", "GSE79416"], "resolved_accessions": ["GSE167110", "GSE79416"], "statements": ["All data generated or analyzed during the course of this study are included in the published article and its supplementary documents."], "basis": "The paper names GSE167110, GSE79416 in Gene Expression Omnibus (GEO) and the accession resolves in the repository. No dataset was downloaded and no licence was read."}}, "can_run": {"dsl_operations": ["normalise_counts", "differential_expression_fit", "fdr_bh"], "assets": ["source:pmid:36566220", "data:geo:GSE167110"], "steps": ["run_analysis"]}, "must_build": {"dsl_operations": [], "converters": [], "readers": [], "capabilities": [], "data_gaps": ["data:geo:GSE167110"]}, "gaps": []}, "review": {"state": "partial", "machine_critique": {"kind": "extraction_review", "reader": {"model": "MiniMax-M3.1-Flash", "provider": "minimax", "provider_family": "minimax", "read_at": "2026-11-01T18:21:37Z", "independence": "single_reader"}, "evidence_check": {"method": "verbatim", "availability": "verbatim"}, "calls": 5, "usage": {"prompt_tokens": 13184, "completion_tokens": 1338, "cached_tokens": 264}}, "human_critique": null, "stages": [{"id": "extraction", "state": "complete", "reader": "MiniMax-M3.1-Flash", "provider": "minimax"}], "gaps": ["review.independent_reader", "review.machine_critique", "review.human_critique"]}, "execution": {"state": "absent", "status": "not_run", "mode": null, "runs": [], "gaps": ["execution.not_run"]}, "provenance": {"entities": [{"id": "pmid:36566220", "type": "paper", "label": "pmid:36566220"}, {"id": "source:pmid:36566220", "type": "manuscript", "label": "PMID 36566220 full text"}, {"id": "data:geo:GSE167110", "type": "dataset", "label": "GEO GSE167110"}], "activities": [{"id": "activity-reading-sci-2026-11-01", "type": "classification", "used": [{"id": "pmid:36566220", "type": "paper", "label": "pmid:36566220"}, {"id": "source:pmid:36566220", "type": "manuscript", "label": "PMID 36566220 full text"}, {"id": "data:geo:GSE167110", "type": "dataset", "label": "GEO GSE167110"}], "agent": "agent:corpus-assay-engine"}, {"id": "activity-read-36566220", "type": "extraction", "used": ["source:pmid:36566220"], "agent": "agent:minimax-reader"}, {"id": "activity-resolve-GSE167110", "type": "accession_resolution", "used": ["data:geo:GSE167110"], "agent": "agent:corpus-assay-engine"}], "agents": [{"id": "agent:corpus-assay-engine", "type": "software", "label": "Corpus assay engine"}, {"id": "agent:minimax-reader", "type": "software", "label": "MiniMax literature reader"}], "source_run_id": "reading-sci-2026-11-01", "dsl_catalogue_fingerprint": "343fb30756ba6ec3", "capability_table_fingerprint": "2cb3e42f047d2ecf", "snapshot_id": "ba34b49490d9bfaf"}, "lineage": {"parent_id": "pmid:36566220", "parent_hash": "sha256:e84fda9d33394cfe56ab11e8fbe9add696e5a7934b049e0095b576fb32f17dd0", "fork_of": null, "generation": 3}, "gaps": ["asset:data:geo:GSE167110 licence not read", "asset:data:geo:GSE167110 not downloaded", "asset:data:geo:GSE79416 licence not read", "asset:data:geo:GSE79416 not downloaded", "execution.not_run", "method.reader_differs_from_extractor", "review.human_critique", "review.independent_reader", "review.machine_critique"], "assays": [{"role": "data_generation", "definition_id": "apoptosis_assay", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["R2 attenuates neuroinflammation and neuronal apoptosis after subarachnoid hemorrhage through the PI", "BACKGROUND: Neuroinflammation and neuronal apoptosis are closely associated with a poor prognosis", "ated neuroinflammation, and reduced neuronal apoptosis after SAH. Additionally, the protective effe", "y, alleviated neuroinflammation and neuronal apoptosis in vivo and in vitro. CCR2 levels in the CSF"], "order_basis": "paper_text_order:16", "id": "assay:1"}, {"role": "data_generation", "definition_id": "rna_seq", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ptor 2 (CCR2) in SAH. METHODS: Pre-processed RNA-seq transcriptome datasets GSE167110 and GSE7941", "nism of CCR2 after SAH. Statistical analysis RNA-seq data were statistically analyzed using R Stu", "CCR2) in SAH. METHODS: Pre-processed RNA-seq transcriptome datasets GSE167110 and GSE79416 from the Gen"], "order_basis": "paper_text_order:329", "id": "assay:2"}, {"role": "data_generation", "definition_id": "elisa", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["mination, enzyme-linked immunosorbent assay (ELISA), western blotting, and immunofluorescence w", "ospinal fluid (CST) samples were detected by ELISA. RESULTS: Expression of CCR2 was upregulated", "in barrier (BBB) permeability determination, enzyme-linked immunosorbent assay (ELISA), western blotting, and immunofluores"], "order_basis": "paper_text_order:865", "id": "assay:3"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": [", enzyme-linked immunosorbent assay (ELISA), western blotting, and immunofluorescence were performed. An i"], "order_basis": "paper_text_order:908", "id": "assay:4"}, {"role": "data_generation", "definition_id": "ihc", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["sorbent assay (ELISA), western blotting, and immunofluorescence were performed. An in vitro model of SAH was"], "order_basis": "paper_text_order:930", "id": "assay:5"}, {"role": "data_generation", "definition_id": "animal_model", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["lasm of neurons after SAH, both in vitro and in vivo. RS significantly reduced the brain water co", "ression was upregulated in both in vitro and in vivo SAH models. Additionally, inhibition of CCR2", "ted neuroinflammation and neuronal apoptosis in vivo and in vitro. CCR2 levels in the CSF have a", "d in vitro SAH model For the in vitro study, murine HT22 cells were cultured in Dulbecco's modif"], "order_basis": "paper_text_order:1389", "id": "assay:6"}, {"role": "data_generation", "definition_id": "cell_culture", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.77, "evidence": ["r the in vitro study, murine HT22 cells were cultured in Dulbecco's modified Eagle medium (‘DEME’) co"], "order_basis": "paper_text_order:3057", "id": "assay:7"}, {"role": "analysis", "definition_id": "differential_expression", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["normalise_counts", "differential_expression_fit", "fdr_bh"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 1.0, "evidence": ["The datasets of differential RNA-seq transcriptome data in SAH were compared with those of control mouse brain tissue using the DESeq2 software package (version 1.34.0), using R software (https://www.r-project.org/) [14]."], "order_basis": "consumes_earlier_output", "id": "assay:8"}]}