Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Pharmacology of Chinese Material Medica, School of Traditional Chinese Pharmacy, China Pharmaceutical University, 639 Longmian Road, Nanjing, 211198, PR China.
Departments of Neurology, University of California, Davis, School of Medicine and Shriners Hospital, Sacramento, CA, 95817, USA.
J Ethnopharmacol. 2020 Nov 15;262:113161. doi: 10.1016/j.jep.2020.113161. Epub 2020 Jul 27.
Thrombolytic therapy with tissue plasminogen activator (tPA) after ischemic stroke exacerbates blood-brain barrier (BBB) breakdown and leads to hemorrhagic transformation (HT). YiQiFuMai Lyophilized Injection (YQFM) is a modern preparation derived from Sheng-mai San (a traditional Chinese medicine). YQFM attenuates the BBB dysfunction induced by cerebral ischemia-reperfusion injury. However, whether YQFM can suppress tPA-induced HT remains unknown.
We investigated the therapeutic effect of YQFM on tPA-induced HT and explored the underlying mechanisms in vivo and in vitro to improve the safety of tPA use against stroke.
Male C57BL/6J mice were subjected to 45 min of ischemia and 24 h of reperfusion. tPA (10 mg/kg) were infused 2 h after occlusion and YQFM (0.671 g/kg) was injected 2.5 h after occlusion. The in vitro effect of YQFM (100, 200, 400 μg/mL) on tPA (60 μg/mL)-induced dysfunction of the microvascular endothelial barrier in the brain following oxygen-glucose deprivation/reoxygenation (OGD/R) was observed in bEnd.3 cells.
YQFM suppressed tPA-induced high hemoglobin level in the brain, mortality, neurologic severity score, BBB permeability, expression and activation of matrix metalloproteinase (MMP)-9 and MMP-2, and degradation of tight-junction proteins. Furthermore, YQFM significantly blocked tPA-induced brain microvascular endothelial permeability and phosphorylation of Rho-associated kinase (ROCK)1, myosin light chain (MLC), cofilin and p65 in vivo and in vitro.
YQFM suppressed tPA-induced HT by inhibiting cytoskeletal rearrangement linked with ROCK-cofilin/MLC pathways and inhibiting the nuclear factor-kappa B pathway to ameliorate BBB damage caused by tPA.
组织型纤溶酶原激活物（tPA）溶栓治疗会加重缺血性脑卒中后血脑屏障（BBB）的破坏，并导致出血性转化（HT）。益气复脉冻干注射液（YQFM）是一种从生脉散（一种中药）中衍生而来的现代制剂。YQFM 可减轻脑缺血再灌注损伤引起的 BBB 功能障碍。然而，YQFM 是否能抑制 tPA 诱导的 HT 尚不清楚。
本研究旨在探讨 YQFM 对 tPA 诱导的 HT 的治疗作用，并在体内和体外研究其潜在机制，以提高 tPA 治疗脑卒中的安全性。
雄性 C57BL/6J 小鼠接受 45 分钟缺血和 24 小时再灌注。在闭塞后 2 小时给予 tPA（10mg/kg），并在闭塞后 2.5 小时给予 YQFM（0.671g/kg）。观察 YQFM（100、200、400μg/mL）在氧葡萄糖剥夺/复氧（OGD/R）后对 tPA（60μg/mL）诱导的大脑微血管内皮屏障功能障碍的体外作用。
YQFM 抑制 tPA 诱导的大脑中高血红蛋白水平、死亡率、神经严重程度评分、BBB 通透性、基质金属蛋白酶（MMP）-9 和 MMP-2 的表达和激活以及紧密连接蛋白的降解。此外，YQFM 还显著阻断了 tPA 诱导的体内和体外脑微血管内皮通透性以及 Rho 相关激酶（ROCK）1、肌球蛋白轻链（MLC）、丝切蛋白和 p65 的磷酸化。
YQFM 通过抑制与 ROCK-丝切蛋白/MLC 通路相关的细胞骨架重排以及抑制核因子-κB 通路来抑制 tPA 引起的 HT，从而改善 BBB 损伤。
