Yamada Narumi, Nakagawa Shinsuke, Horai Shoji, Tanaka Kunihiko, Deli Maria A, Yatsuhashi Hiroshi, Niwa Masami
Clinical Research Center, National Hospital Organization Nagasaki Medical Center, 2-1001-1 Kubara, Omura 856-8562, Japan; Department of Hepatology, Nagasaki University Graduate School of Biomedical Sciences, 2-1001-1 Kubara, Omura 856-8562, Japan.
Department of Pharmacology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; BBB Laboratory, PharmaCo-Cell Company, Ltd., 1-43 Dejima, Nagasaki 850-0862, Japan.
Microvasc Res. 2014 Mar;92:41-9. doi: 10.1016/j.mvr.2013.12.004. Epub 2013 Dec 23.
The effects of hepatocyte growth factor (HGF) on barrier functions were investigated by a blood-brain barrier (BBB) in vitro model comprising a primary culture of rat brain capillary endothelial cells (RBEC). In order to examine the response of the peripheral endothelial cells to HGF, human umbilical vascular endothelial cells (HUVEC) and human dermal microvascular endothelial cells (HMVEC) were also treated with HGF. HGF decreased the permeability of RBEC to sodium fluorescein and Evans blue albumin, and dose-dependently increased transendothelial electrical resistance (TEER) in RBEC. HGF altered the immunochemical staining pattern of F-actin bands and made ZO-1 staining more distinct on the linear cell borders in RBEC. In contrast, HGF increased sodium fluorescein and Evans blue albumin permeability in HMVEC and HUVEC, and decreased TEER in HMVEC. In HMVEC, HGF reduced cortical actin bands and increased stress fiber density, and increased the zipper-like appearance of ZO-1 staining. Western blot analysis showed that HGF significantly increased the amount of ZO-1 and VE-cadherin. HGF seems to act on the BBB to strengthen BBB integrity. These findings indicated that cytoskeletal rearrangement and cell-cell adhesion, such as through VE-cadherin and ZO-1, are candidate mechanisms for the influence of HGF on the BBB. The possibility that HGF has therapeutic significance in protecting the BBB from damage needs to be considered.
利用包含大鼠脑毛细血管内皮细胞（RBEC）原代培养物的血脑屏障（BBB）体外模型，研究了肝细胞生长因子（HGF）对屏障功能的影响。为了检测外周内皮细胞对HGF的反应，人脐静脉内皮细胞（HUVEC）和人真皮微血管内皮细胞（HMVEC）也用HGF进行了处理。HGF降低了RBEC对荧光素钠和伊文思蓝白蛋白的通透性，并剂量依赖性地增加了RBEC的跨内皮电阻（TEER）。HGF改变了F-肌动蛋白带的免疫化学染色模式，并使RBEC线性细胞边界上的ZO-1染色更明显。相反，HGF增加了HMVEC和HUVEC中荧光素钠和伊文思蓝白蛋白的通透性，并降低了HMVEC中的TEER。在HMVEC中，HGF减少了皮质肌动蛋白带并增加了应力纤维密度，并增加了ZO-1染色的拉链样外观。蛋白质印迹分析表明，HGF显著增加了ZO-1和血管内皮钙黏蛋白的量。HGF似乎作用于血脑屏障以加强血脑屏障的完整性。这些发现表明，细胞骨架重排和细胞间黏附，如通过血管内皮钙黏蛋白和ZO-1，是HGF影响血脑屏障的候选机制。需要考虑HGF在保护血脑屏障免受损伤方面具有治疗意义的可能性。
