血管平滑肌细胞LRRC8A Cl-通道的生物学特性与功能及与ClC-3 Cl-通道的关系和药物干预的研究

容积调节Cl- 通道 (VRCC) 的分子基础一直存在着争论, ,我们证实ClC-3 是血管平滑肌细胞(VSMCs) VRCC的有效组成成份,参与脑血管平滑肌细胞增殖,凋亡及脑血管重构. 近有学者提出LRRC8A 是VRCC的有效组成成份。但是在不同的细胞上仍有争论。对血管平滑肌细胞上LRRC8A特性和功能一无所知。本项目拟在原有基础上，通过LRRC8A基因敲除鼠整体与基因沉默及突变等离体实验，采用斑片钳, 荧光影像测定单细胞Cl-跨膜运动及分子生物学等技术，探讨LRRC8A Cl-通道在血管平滑肌细胞上的生物学特性, 与ClC-3容积调节Cl-通道之间关系；LRRC8A-Cl-通道是否干预细胞增殖与凋亡参与脑血管重构，及药物对之干预作用。为揭示LRRC8A在血管平滑肌细胞的生物学特性及LRRC8A作为新靶点寻求防治高血压脑血管重构及脑卒中新药提供新的科学依据和线索。

For a long time， volume regulated Cl- channel (VRCC) on the molecular identification is kept in debate. We have confirmed that ClC-3 is an effective component of VRCC in vascular smooth muscle cells, and participates in cerebral vascular smooth muscle cell proliferation, apoptosis and remodeling. Recent scholars have put forward LRRC8A is an effective component of VRCC. But not confirmed on different cells. LRRC8A features and functions are not known in vascular smooth muscle cells. To this end, we intend to work on the basis of the original, through LRRC8A knockout mice in vivo experiment, gene silencing and mutation in vitro, using patch clamp single ion channel recording, dynamic determination of the fluorescence images of Cl- transmembrane movement and molecular biology technology, to explore the biological characteristics of LRRC8A Cl- channel in vascular smooth muscle cells , the relationship between ClC-3 volume regulated and LRRC8A Cl- channels, and LRRC8A channel regulated signal pathway; to investigate whether the LRRC8A-Cl- channel involves into cell proliferation, apoptosis and cerebrovascular remodeling, as well as intervention of drugs. In order to reveal the biological characteristics and functions of LRRC8A in vascular smooth muscle cells, and provide new scientific basis and clues for LRRC8A as a new target for the prevention and treatment of hypertensive cerebral vascular remodeling and stroke.