Li Limin, Lou Xiaoli, Zhang Kunlun, Yu Fangping, Zhao Yingchun, Jiang Ping
Department of Neurology, The Affiliated Shanghai Songjiang Central Hospital of Shanghai Jiao Tong University, Central Hospital of Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Transl Neurosci. 2020 Apr 20;11(1):75-86. doi: 10.1515/tnsci-2020-0100. eCollection 2020.
The aim of this study was to investigate the neuroprotective effects of hydrochloride fasudil (HF) in rats following intracerebral hemorrhage (ICH).
Male Wistar rats were randomly divided into four groups: normal, sham-operated, ICH, and ICH/HF. ICH was induced by injection of non-anticoagulant autologous arterial blood into the right caudate nucleus. The levels of Rho-associated protein kinase 2 (ROCK2) mRNA and protein around the site of the hematoma were measured by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA), respectively. The levels of interleukin-6 and tumor necrosis factor-α in serum were detected by ELISA. The inflammatory cells and changes in the neuronal morphology around the hematoma were visualized using hematoxylin and eosin and Nissl staining. Brain edema was measured by comparing wet and dry brain weights.
Following ICH, the levels of ROCK2 were significantly increased from day 1 to day 7. The levels of ROCK2 were significantly lower in rats treated with HF than in controls. The levels of inflammatory cytokines and brain water content were significantly higher in rats treated with HF than in controls. Administration of HF significantly reduced the levels of inflammatory cytokines and brain water content from day 1 to day 7. In the acute phase of ICH, a large number of neutrophils infiltrated the perihematomal areas. In comparison with the ICH group, the ICH/HF group showed markedly fewer infiltrating neutrophils on day 1. Nissl staining showed that ICH caused neuronal death and loss of neurons in the perihematomal areas at all time points and that treatment with HF significantly attenuated neuronal loss.
HF exerts neuroprotective effects in ICH rats by inhibiting the expression of ROCK2, reducing neutrophil infiltration and production of inflammatory cytokines, decreasing brain edema, and attenuating loss of neurons.
本研究旨在探讨盐酸法舒地尔（HF）对脑出血（ICH）大鼠的神经保护作用。
雄性Wistar大鼠随机分为四组：正常组、假手术组、ICH组和ICH/HF组。通过向右侧尾状核注射非抗凝自体动脉血诱导脑出血。分别采用定量实时聚合酶链反应和酶联免疫吸附测定（ELISA）法检测血肿部位周围Rho相关蛋白激酶2（ROCK2）mRNA和蛋白水平。采用ELISA法检测血清中白细胞介素-6和肿瘤坏死因子-α水平。用苏木精-伊红染色和尼氏染色观察血肿周围的炎性细胞和神经元形态变化。通过比较脑湿重和干重测量脑水肿。
脑出血后，ROCK2水平从第1天到第7天显著升高。HF治疗的大鼠ROCK2水平显著低于对照组。HF治疗的大鼠炎性细胞因子水平和脑含水量显著高于对照组。从第1天到第7天，给予HF显著降低了炎性细胞因子水平和脑含水量。在脑出血急性期，大量中性粒细胞浸润血肿周围区域。与ICH组相比，ICH/HF组在第1天浸润的中性粒细胞明显减少。尼氏染色显示，脑出血在所有时间点均导致血肿周围区域神经元死亡和神经元丢失，而HF治疗显著减轻了神经元丢失。
HF通过抑制ROCK2表达、减少中性粒细胞浸润和炎性细胞因子产生、减轻脑水肿以及减轻神经元丢失，对脑出血大鼠发挥神经保护作用。
