title: ATP Induces Disruption of Tight Junction Proteins via IL-1 Beta-Dependent MMP-9 Activation of Human Blood-Brain BarrierIn Vitro.
abstract: Disruption of blood-brain barrier (BBB) follows brain trauma or central nervous system (CNS) stress. However, the mechanisms leading to this process or the underlying neural plasticity are not clearly known. We hypothesized that ATP/P2X7R signaling regulates the integrity of BBB. Activation of P2X7 receptor (P2X7R) by ATP induces the release of interleukin-1β(IL-1β), which in turn enhances the activity of matrix metalloproteinase-9 (MMP-9). Degradation of tight junction proteins (TJPs) such as ZO-1 and occludin occurs, which finally contributes to disruption of BBB. A contact coculture system using human astrocytes and hCMEC/D3, an immortalized human brain endothelial cell line, was used to mimic BBBin vitro. Permeability was used to evaluate changes in the integrity of TJPs. ELISA, Western blot, and immunofluorescent staining procedures were used. Our data demonstrated that exposure to the photoreactive ATP analog, 3'-O-(4-benzoyl)benzoyl adenosine 5'-triphosphate (BzATP), induced a significant decrease in ZO-1 and occludin expression. Meanwhile, the decrease of ZO-1 and occludin was significantly attenuated by P2X7R inhibitors, as well as IL-1R and MMP antagonists. Further, the induction of IL-1βand MMP-9 was closely linked to ATP/P2X7R-associated BBB leakage. In conclusion, our study explored the mechanism of ATP/P2X7R signaling in the disruption of BBB following brain trauma/stress injury, especially focusing on the relationship with IL-1βand MMP-9.
authors: Department of Neurosurgery, Chinese PLA General Hospital, 28 Fuxing Road, Haidian District, Beijing 100853, China; Department of Neurosurgery, 2nd Affiliated Hospital of Fujian Medical University, 34 Zhongshan Northern Road, Quanzhou 362000, China.
authors: Fuxing
authors: 0000-0002-5220-342X
authors: F
authors: Yang
authors: Department of Neurosurgery, Chinese PLA General Hospital, 28 Fuxing Road, Haidian District, Beijing 100853, China.
authors: Kai
authors: K
authors: Zhao
authors: Department of Neurosurgery, Chinese PLA General Hospital, 28 Fuxing Road, Haidian District, Beijing 100853, China; Medical College, Nankai University, 94 Weijin Road, Tianjin 300071, China.
authors: Xiufeng
authors: 0000-0002-5801-1613
authors: X
authors: Zhang
authors: Department of Neurosurgery, Chinese PLA General Hospital, 28 Fuxing Road, Haidian District, Beijing 100853, China.
authors: Jun
authors: 0000-0001-6439-0608
authors: J
authors: Zhang
authors: Department of Neurosurgery, Chinese PLA General Hospital, 28 Fuxing Road, Haidian District, Beijing 100853, China.
authors: Bainan
authors: 0000-0001-5914-4939
authors: B
authors: Xu
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D018920:Coculture Techniques; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D058920:Purinergic P2X Receptor Antagonists / Q000494:pharmacology; D058486:Receptors, Purinergic P2X7 / Q000378:metabolism*; D062725:Tight Junction Proteins / Q000378:metabolism*
chemical_list: D053583:Interleukin-1beta; C546468:P2RX7 protein, human; D058920:Purinergic P2X Receptor Antagonists; D058486:Receptors, Purinergic P2X7; D062725:Tight Junction Proteins; D000255:Adenosine Triphosphate; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9