Shaoyu Guan, Ruichen Jiang, Xudong Wang, Tong Chen, Ping Yi, Tian Li, Teng Ma, Fang Wang
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After bioinformatics analysis of the transcriptome in vivo, qRT-PCR of miR-297c-5p was conducted and a dual-luciferase reporter assay was used to verify the target protein, occludin, which was confirmed by hippocampal insertion using guide cannulas and microinfection of RNA oligos.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>A 3-h deprivation of OGD of b. End3 cells resulted in noticeable reductions in the level of occludin and ZO-1. However, administration of LSZ (0.1 μM) effectively restored these decreases. In normal mice, administration of LSZ (25 mg/kg, i.p., once daily for 9 days) resulted in a notable reduction in miR-297c-5p. In the middle cerebral artery occlusion (MCAO) mouse model, increased miR-297c-5p was also reversed by LSZ administration. Bioinformatics analysis revealed one of the targets of miR-297c-5p includes occludin. MiR-297c-5p was found to directly target occludin in the dual-luciferase reporter assay. Transfection of miR-297c-5p agomir into b. End3 cells resulted in a significant reduction in the level of occludin, while transfection of antagomir led to an increase in occludin. Besides, stereotaxic injection of AAV-miR-297c-5p into the hippocampus reduced occludin level in vivo. Ultimately, hippocampal microinfection of RNA oligos provided a confirmation that miR-297c-5p was downregulated by LSZ in MCAO mice with up-regulated occludin expression.</p>\n </section>\n \n <section>\n \n <h3> Conclusion</h3>\n \n <p>In conclusion, the present findings provide new insights into regulating occludin by LSZ through downregulation of miR-297c-5p.</p>\n </section>\n </div>","PeriodicalId":154,"journal":{"name":"CNS Neuroscience & Therapeutics","volume":"31 5","pages":""},"PeriodicalIF":4.8000,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/cns.70367","citationCount":"0","resultStr":"{\"title\":\"Ligustrazine Alleviates Blood–Brain Barrier Damage by Downregulating Expression of miR-297c-5p\",\"authors\":\"Shaoyu Guan, Ruichen Jiang, Xudong Wang, Tong Chen, Ping Yi, Tian Li, Teng Ma, Fang Wang\",\"doi\":\"10.1111/cns.70367\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Objective</h3>\\n \\n <p>Ligustrazine (LSZ), an ingredient of <i>Ligusticum chuanxiong</i>, has long been used to treat neurovascular diseases in China. This study investigates its protective effects for the impairment of the blood–brain barrier (BBB) and the underlying mechanisms.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>In this study, the impacts of LSZ on the BBB function were firstly assessed in b. End3 cells in vitro. Oxygen–glucose deprivation (OGD) served as an injury factor and western blot (WB) analyzed the expressions of occludin and ZO-1, two tight junction proteins (TJs), essential for maintaining the integrity of the BBB. After bioinformatics analysis of the transcriptome in vivo, qRT-PCR of miR-297c-5p was conducted and a dual-luciferase reporter assay was used to verify the target protein, occludin, which was confirmed by hippocampal insertion using guide cannulas and microinfection of RNA oligos.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>A 3-h deprivation of OGD of b. End3 cells resulted in noticeable reductions in the level of occludin and ZO-1. 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Ligustrazine Alleviates Blood–Brain Barrier Damage by Downregulating Expression of miR-297c-5p
Objective
Ligustrazine (LSZ), an ingredient of Ligusticum chuanxiong, has long been used to treat neurovascular diseases in China. This study investigates its protective effects for the impairment of the blood–brain barrier (BBB) and the underlying mechanisms.
Methods
In this study, the impacts of LSZ on the BBB function were firstly assessed in b. End3 cells in vitro. Oxygen–glucose deprivation (OGD) served as an injury factor and western blot (WB) analyzed the expressions of occludin and ZO-1, two tight junction proteins (TJs), essential for maintaining the integrity of the BBB. After bioinformatics analysis of the transcriptome in vivo, qRT-PCR of miR-297c-5p was conducted and a dual-luciferase reporter assay was used to verify the target protein, occludin, which was confirmed by hippocampal insertion using guide cannulas and microinfection of RNA oligos.
Results
A 3-h deprivation of OGD of b. End3 cells resulted in noticeable reductions in the level of occludin and ZO-1. However, administration of LSZ (0.1 μM) effectively restored these decreases. In normal mice, administration of LSZ (25 mg/kg, i.p., once daily for 9 days) resulted in a notable reduction in miR-297c-5p. In the middle cerebral artery occlusion (MCAO) mouse model, increased miR-297c-5p was also reversed by LSZ administration. Bioinformatics analysis revealed one of the targets of miR-297c-5p includes occludin. MiR-297c-5p was found to directly target occludin in the dual-luciferase reporter assay. Transfection of miR-297c-5p agomir into b. End3 cells resulted in a significant reduction in the level of occludin, while transfection of antagomir led to an increase in occludin. Besides, stereotaxic injection of AAV-miR-297c-5p into the hippocampus reduced occludin level in vivo. Ultimately, hippocampal microinfection of RNA oligos provided a confirmation that miR-297c-5p was downregulated by LSZ in MCAO mice with up-regulated occludin expression.
Conclusion
In conclusion, the present findings provide new insights into regulating occludin by LSZ through downregulation of miR-297c-5p.
期刊介绍：
CNS Neuroscience & Therapeutics provides a medium for rapid publication of original clinical, experimental, and translational research papers, timely reviews and reports of novel findings of therapeutic relevance to the central nervous system, as well as papers related to clinical pharmacology, drug development and novel methodologies for drug evaluation. The journal focuses on neurological and psychiatric diseases such as stroke, Parkinson’s disease, Alzheimer’s disease, depression, schizophrenia, epilepsy, and drug abuse.
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