{
  "abstract": "Background The increased permeability of the blood-brain barrier (BBB) is a critical contributor to the high mortality following ischaemic stroke. However, the mechanisms regulating BBB integrity remain poorly understood. Leucine-rich repeat-containing 8A (LRRC8A) is a chloride channel critical for cellular volume homeostasis and plays a key role in regulating neuronal injury during ischaemia. However, its impact on BBB function is currently unclear.Methods A transient middle cerebral artery occlusion model was established to investigate the impact of LRRC8A on BBB integrity. Laser speckle contrast imaging was used to monitor cortical blood flow. Primary mouse and human brain microvascular endothelial cells (m/hBMVECs) were subjected to oxygen-glucose deprivation (OGD) and re-oxygenation for varying durations. Patch-clamp recordings were performed to measure volume-regulated chloride currents. Immunostaining was conducted to evaluate protein expression. Cell permeability was evaluated with transwell assay.Results LRRC8A deletion in endothelial cells ameliorates the infarct area and mitigates BBB leakage. Ischaemia dramatically upregulates the expression of LRRC8A in endothelial cells, concurrently downregulating tight junction proteins. OGD exposure augments the VRCC current mediated by LRRC8A in BMVECs. In contrast, inhibiting LRRC8A promotes the expression of ZO-1 and VE-cadherin, thereby preserving the integrity of endothelial cells. With-no-lysine kinase 1 (WNK1) inhibition contributes to LRRC8A-induced BBB damage post-ischaemic stroke. Eupatorin, a newly identified LRRC8A inhibitor, exerts neuroprotective effects against ischaemic stroke.Conclusions LRRC8A in BMVECs plays a pivotal role in modulating BBB integrity, a process regulated by WNK1. As an LRRC8A inhibitor, Eupatorin holds the potential for ischaemic stroke therapy.",
  "authors": [
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Yanfei Tian"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Yan Wang"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Yupeng Zhao"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Chang Liu"
    },
    {
      "affiliations": [
        "Core Facilities and Centers, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Xiaoyu Zhang"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Yan Zhang"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Zhenghui Wu"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Yue Kong"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Bo Wang"
    },
    {
      "affiliations": [
        "Core Facilities and Centers, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Huaxing Zhang"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China",
        "The Key Laboratory of Neural and Vascular Biology; Ministry of Education, Shijiazhuang, China"
      ],
      "name": "Xiaona Du"
    },
    {
      "affiliations": [
        "Department of Pharmacology, The Key Laboratory of New Drug Pharmacology and Toxicology, Hebei Medical University, Shijiazhuang, Hebei, China",
        "The Key Laboratory of Neural and Vascular Biology; Ministry of Education, Shijiazhuang, China",
        "Department of Psychiatry, The First Hospital of Hebei Medical University, Mental Health Institute of Hebei Medical University, Shijiazhuang, Hebei, China",
        "Collaborative Innovation Center of Hebei Province for Mechanism, Diagnosis and Treatment of Neuropsychiatric Diseases, Shijiazhuang, Hebei, China"
      ],
      "name": "Hailin Zhang"
    },
    {
      "affiliations": [
        "The Key Laboratory of Neural and Vascular Biology; Ministry of Education, Shijiazhuang, China",
        "Department of Medical and Pharmaceutical Informatics, Hebei Medical University, Shijiazhuang, Hebei, China"
      ],
      "name": "Huiran Zhang"
    }
  ],
  "title": "LRRC8A in endothelial cells contributes to the aberrant blood-brain barrier integrity in ischaemic stroke",
  "uid": "6b8ba181-ce37-56e6-87db-1ae349029619"
}