Hyaluronic Acid binding protein 2 is a novel regulator of vascular integrity.
Hyaluronic Acid binding protein 2 is a novel regulator of vascular integrity.
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DOI：
10.1161/atvbaha.109.200451
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发表时间：
2010-03
期刊：
影响因子：
--
通讯作者：
Singleton PA
中科院分区：
文献类型：
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作者：
Mambetsariev N;Mirzapoiazova T;Mambetsariev B;Sammani S;Lennon FE;Garcia JG;Singleton PA
我们评估了细胞外丝氨酸蛋白酶，透明质酸结合蛋白2（HABP 2），在血管屏障调节中的作用。采用免疫印迹和免疫组织化学方法，观察了脂多糖（LPS）诱导小鼠肺内皮细胞和体外培养的人肺微血管内皮细胞（HPMVEC）表达HABP 2。高分子量透明质酸（HMW-HA，~ 100万Da）降低HPMVEC中HABP 2蛋白的表达，并降低纯化的HABP 2酶活性，而低分子量HA（LMW-HA，~2,500 Da）增加这些活性。HABP 2的聚阴离子结合结构域（PABD）的肽抑制了LMW-HA对HABP 2活性的影响，但HMW-HA没有。沉默（siRNA）HABP 2表达增强HMW-HA诱导的EC屏障增强和抑制LPS和LMW-HA介导的EC屏障破坏，结果与HABP 2的过表达逆转。沉默PAR受体1和3、RhoA或ROCK表达减弱LPS、LMW-HA和HABP 2介导的EC屏障破坏。利用急性肺损伤的小鼠模型，我们观察到HABP 2的血管沉默（siRNA）显著降低了LPS和呼吸机诱导的肺血管高通透性。HABP 2通过激活PAR受体/RhoA/ROCK信号负调节血管完整性，并代表了血管通透性增加综合征的潜在有用的治疗靶点。
We evaluated the role of the extracellular serine protease, Hyaluronic Acid Binding Protein 2 (HABP2), in vascular barrier regulation. Using immunoblot and immunohistochemical analysis, we observed that lipopolysaccharide (LPS)-induces HABP2 expression in murine lung endothelium in vivo and in human pulmonary microvascular endothelial cell (HPMVEC) in vitro. High molecular weight hyaluronan (HMW-HA, ~1 million Da) decreased HABP2 protein expression in HPMVEC and decreased purified HABP2 enzymatic activity whereas low MW HA (LMW-HA, ~2,500 Da) increased these activities. The effects of LMW-HA on HABP2 activity, but not HMW-HA, were inhibited with a peptide of the polyanion binding domain (PABD) of HABP2. Silencing (siRNA) HABP2 expression augmented HMW-HA-induced EC barrier enhancement and inhibited LPS and LMW-HA-mediated EC barrier disruption, results which were reversed with overexpression of HABP2. Silencing PAR receptors 1 and 3, RhoA or ROCK expression attenuated LPS, LMW-HA and HABP2-mediated EC barrier disruption. Utilizing murine models of acute lung injury, we observed that LPS- and ventilator-induced pulmonary vascular hyper-permeability were significantly reduced with vascular silencing (siRNA) of HABP2. HABP2 negatively regulates vascular integrity via activation of PAR receptor/RhoA/ROCK signaling and represents a potentially useful therapeutic target for syndromes of increased vascular permeability.