Cao Guo-Sheng, Chen Hong-Lin, Zhang Yuan-Yuan, Li Fang, Liu Chun-Hua, Xiang Xiang, Qi Jin, Chai Cheng-Zhi, Kou Jun-Ping, Yu Bo-Yang
Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing, PR China.
J Ethnopharmacol. 2016 May 13;183:18-28. doi: 10.1016/j.jep.2016.02.028. Epub 2016 Feb 23.
YiQiFuMai Powder Injection (YQFM) is a modern preparation derived from Sheng-mai San, a traditional Chinese prescription, widely used for the treatment of cardiovascular and cerebrovascular diseases. However, its potential molecular mechanism remains unclear.
The present study was designed to observe the effect of YQFM on oxygen-glucose deprivation (OGD)-induced the brain microvascular endothelial barrier dysfunction and to explore the underlying pathways in vitro.
A mouse brain microvascular endothelial cell line (bEnd.3) was subjected to OGD (2-9h) to examine the efficacy and molecular mechanisms in the presence or absence of YQFM (100, 200 and 400 μg/ml).
The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Trans-endothelial electrical resistance (TEER) assays demonstrated that treatment with YQFM increased the cell viability and TEER value, decreased even blue (EB) albumin leakage after OGD in bEnd.3 cells. Western blotting and immunofluorescence staining showed that YQFM reduced the breakage and translocation of Zonula occludens-1 (ZO-1) and claudin-5 after 4h of OGD and decreased the expression of these proteins after 9h of OGD. Moreover, YQFM significantly inhibited the expression, phosphorylation and nuclear translocation of NF-κB/p65 and decreased the expression of intercellular adhesionmolecule-1 (ICAM-1) and cyclooxygenase (COX-2) as well as production of nitric oxide (NO). In addition, real time-PCR results revealed that YQFM suppressed the mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) after 4h of OGD. Furthermore, YQFM markedly inhibited both the phosphorylation of myosin light chain (MLC) and cytoskeletal reorganization and reduced the expression of cleaved-ROCK1 in bEnd.3 cells subjected to OGD.
These findings suggest that YQFM ameliorates the OGD-induced brain microvascular endothelial cell barrier disruption associated with the NF-κB/p65 and ROCK1/MLC signaling pathways. These data provide new insights into the use of this preparation for treating cerebrovascular diseases.
益气复脉粉针剂（YQFM）是一种源自传统中药生脉散的现代制剂，广泛用于治疗心脑血管疾病。然而，其潜在的分子机制仍不清楚。
本研究旨在观察益气复脉粉针剂对氧糖剥夺（OGD）诱导的脑微血管内皮屏障功能障碍的影响，并在体外探索其潜在机制。
用氧糖剥夺（2 - 9小时）处理小鼠脑微血管内皮细胞系（bEnd.3），以检测在有或无异气复脉粉针剂（100、200和400μg/ml）情况下的疗效和分子机制。
3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）和跨内皮电阻（TEER）检测结果表明，益气复脉粉针剂处理可提高bEnd.3细胞活力和TEER值，减少OGD后伊文思蓝（EB）白蛋白渗漏。蛋白质免疫印迹法和免疫荧光染色显示，OGD 4小时后，益气复脉粉针剂可减少紧密连接蛋白1（ZO-1）和闭合蛋白5的断裂和转位，OGD 9小时后可降低这些蛋白的表达。此外，益气复脉粉针剂可显著抑制NF-κB/p65的表达、磷酸化和核转位，并降低细胞间黏附分子-1（ICAM-1）和环氧化酶（COX-2）的表达以及一氧化氮（NO）的产生。另外，实时定量聚合酶链反应结果显示，OGD 4小时后，益气复脉粉针剂可抑制肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的mRNA水平。此外，益气复脉粉针剂可显著抑制肌球蛋白轻链（MLC）的磷酸化和细胞骨架重组，并降低OGD处理的bEnd.3细胞中裂解的ROCK1的表达。
这些发现表明益气复脉粉针剂可改善OGD诱导的与NF-κB/p65和ROCK1/MLC信号通路相关的脑微血管内皮细胞屏障破坏。这些数据为该制剂用于治疗脑血管疾病提供了新的见解。
