Bai Shi, Sun Yayi, Wu Lijuan, Wu Zhongmin, Fang Marong
School of Medicine, Taizhou University, Taizhou 318000, China; Institute of Neuroscience, Zhejiang University School of Medicine, Hangzhou 310058, China.
Institute of Neuroscience, Zhejiang University School of Medicine, Hangzhou 310058, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2016 May 25;45(5):493-500. doi: 10.3785/j.issn.1008-9292.2016.09.07.
To investigate the effects of triptolide on inflammation and apoptosis induced by focal cerebral ischemia/reperfusion in rats. The rat model of focal cerebral ischemia/reperfusion injury was established according to Longa's method. A total of 80 SD rats were randomly divided into 5 groups:normal control, sham group, DMSO group, middle cerebral artery occlusion (MCAO) group, and MCAO with tripolide treatment group. TTC staining was used to examine the site and volume of cerebral infarction, and Longa score was employed for neurological disorders measurement. Number of astrocytes was measured by fluorescence staining, and neuronal apoptosis was determined by TUNEL staining. The expressions of inducible nitric oxide synthase(iNOS), cyclooxygenase 2(COX-2) and NF-κB proteins were detected by immunohistochemistry, and the expression of iNOS, COX-2 mRNA was detected by real-time PCR. Compared with DMSO group and MCAO group, brain edema was improved (80.03±0.46)% (<0.05), infarct volume was reduced (8.3±1.4)% (<0.01), Longa score was decreased (1.38±0.20, <0.05) in triptolide treatment group. Meanwhile triptolide also dramatically reduced the number of GFAP-positive astrocytes (<0.05), alleviated protein expression of COX-2 (91.67±1.31), iNOS (95.24±5.07) and NF-κB (75.03±2.06) triggered by MCAO (all <0.05), and induced a down-regulation of cell apoptosis as showed by TUNEL assay (64.15±3.52, <0.05). Triptolide can reduce the cerebral infarction volume, attenuate brain edema and ameliorate the neurological deficits induced by cerebral ischemia-reperfusion injury rats, indicating that it might be used as a potential anti-inflammatory agent.
探讨雷公藤甲素对大鼠局灶性脑缺血/再灌注诱导的炎症和细胞凋亡的影响。按照Longa法建立大鼠局灶性脑缺血/再灌注损伤模型。将80只SD大鼠随机分为5组：正常对照组、假手术组、二甲基亚砜（DMSO）组、大脑中动脉闭塞（MCAO）组和雷公藤甲素治疗的MCAO组。采用TTC染色检测脑梗死部位和体积，采用Longa评分评估神经功能障碍。通过荧光染色测量星形胶质细胞数量，采用TUNEL染色确定神经元凋亡。采用免疫组化检测诱导型一氧化氮合酶（iNOS）、环氧化酶2（COX-2）和核因子κB（NF-κB）蛋白的表达，采用实时聚合酶链反应检测iNOS、COX-2 mRNA的表达。与DMSO组和MCAO组相比，雷公藤甲素治疗组脑水肿改善（80.03±0.46）%（P<0.05），梗死体积减小（8.3±1.4）%（P<0.01），Longa评分降低（1.38±0.20，P<0.05）。同时，雷公藤甲素还显著减少了胶质纤维酸性蛋白（GFAP）阳性星形胶质细胞数量（P<0.05），减轻了MCAO引发的COX-2（91.67±1.31）、iNOS（95.24±5.07）和NF-κB（75.03±2.06）蛋白表达（均P<0.05），并如TUNEL检测所示诱导细胞凋亡下调（64.15±3.52，P<0.05）。雷公藤甲素可减小脑梗死体积，减轻脑水肿，改善脑缺血-再灌注损伤大鼠的神经功能缺损，表明其可能作为一种潜在的抗炎药物。
