积雪草苷通过激活PI3K/Akt信号通路缓解局灶性脑梗死大鼠脑组织损伤和诱导血管新生  

Asiaticoside alleviates brain tissue injury and induces angiogenesis of focal cerebral infraction rats via activating PI3K/Akt signaling pathway

作　　者：李磊[1] 高绚照[1] 马连萍[1] 苗成[1] 杨惠 LI Lei;GAO Xuan-Zhao;MA Lian-Ping;MIAO Cheng;YANG Hui(Department of Internal Medicine-Neurology,Xinxiang Central Hospital,Xinxiang 453000,China)

机构地区：[1]新乡市中心医院神经内科

出　　处：《中国免疫学杂志》2019年第17期2068-2072,2078,共6页Chinese Journal of Immunology

基　　金：河南省科技厅重点基金资助项目(XZ5689P90)

摘　　要：目的:探究积雪草苷(Asiat)对局灶性脑梗死大鼠脑组织损伤和血管新生的作用和机制。方法:将60只雄性SD大鼠随机分为健康对照(Ctrl)组、大脑中动脉阻塞(MCAO)组、Asiat(10mg/kg)组、Asiat(20mg/kg)组和Asiat(50mg/kg)组。除Ctrl组外,其余组大鼠采用线栓法复制大鼠MCAO模型,Asiat(10、20、50mg/kg)组分别腹腔注射10、20和50mg/kg的Asiat。7d后处死大鼠,摘取脑组织并收集血液,用HE染色检测脑组织损伤,Tunnel染色检测细胞凋亡,ELISA检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)和一氧化氮(NO)浓度,免疫印迹检测Ki67、Caspase-3、Bcl-2、Bax、血管内皮生长因子(VEGF)、VEGF受体2(VEGFR2)、磷脂酰肌醇-3-羟激酶(PI3K)、p-PI3K、Akt和p-Akt的表达。结果:与Ctrl组比较,MCAO组大鼠脑组织损伤加重;与MCAO组比较,Asiat(10、20、50mg/kg)组大鼠脑组织损伤有所减轻,并具有量效关系;MCAO组大鼠脑组织细胞凋亡率、Caspase-3和Bax表达水平与Ctrl组比较明显升高,Ki67和Bcl-2表达明显被抑制;Asiat(10、20、50mg/kg)组细胞凋亡率、Caspase-3和Bax表达水平与MCAO组比较明显降低,Ki67和Bcl-2表达水平明显升高;同时,与Ctrl组比较,MCAO组大鼠血清NO和MDA浓度明显升高,SOD浓度显著降低;与MCAO组比较,Asiat(10、20、50mg/kg)组大鼠血清NO和MDA浓度显著降低,SOD浓度显著升高;此外,Asiat(10、20、50mg/kg)还能促进MCAO模型大鼠VEGF和VEGFR2的表达,升高p-PI3K/PI3K和p-Akt/Akt的比值。结论:Asiat能减轻脑梗死大鼠脑损伤并促进血管新生,作用机制可能与诱导PI3K/Akt通路激活有关。Objective: To investigate the effects and mechanism of asiaticoside on brain tissue injury and angiogenesis of cerebral infraction rats. Methods: 60 male rats were divided into control,middle cerebral artery occlusion,Asiat(10 mg/kg),Asiat (20 mg/kg) and Asiat(50 mg/kg) group,and rats were treated with middle cerebral artery occlusion besides the rats in Ctrl group.Rats in Asiat(10,20,50 mg/kg) were intraperitoneally injected with 10,20,50 mg/kg Asiat respectively.After treating with Asiat for 7 d,rats were sacrificed,the brain tissues and blood were collected. HE staining was performed for brain injury,cell apoptosis was determined by Tunnel assay,the concentrations of superoxide dismutase(SOD),malonaldehyde(MDA)and nitric oxide(NO)were measured by ELISA assay.The protein levels of Ki67,Caspase-3,Bcl-2,Bax,vascular endothelial growth factor (VEGF),VEGF receptor 2 (VEGFR2),phosphatidylinositol 3-hydroxy kinase(PI3K),p-PI3K,Akt and p-Akt were measured by Western blot. Results: Compared with Ctrl group,the brain injury of rats in MCAO group was aggravated;compared with MCAO group,the injury of rats in Asiat(10,20,50 mg/kg) group was attenuated and coupled with dose-effect relationship.The cell apoptosis rate and the expression levels of Caspase-3 and Bax in MCAO group were increased compared with Ctrl group,but the expressions of Ki67 and Bcl-2 were inhibited significantly;the cell apoptosis rate and the expression levels of Caspase-3 and Bax in Asiat(10,20,50 mg/kg) group were decreased compared with MCAO group,and the levels of Ki67 and Bcl-2 were increased markedly.Meanwhile,compared with Ctrl group,the concentrations of NO and MDA were up-regulated in MCAO group,the concentration of SOD was decreased notably;compared with MCAO group;the concentrations of NO and MDA were down-regulated in Asiat(10,20,50 mg/kg)group,the concentration of SOD was increased notably. In addition,Asiat(10,20,50 mg/kg) induced the expressions of VEGF and VEGFR2 of model rats,and increased the ratio of p-PI3K/PI3K and p-Akt/Akt. Conclus

关 键 词：积雪草苷 局灶性脑梗死 血管新生 VEGF PI3K/AKT通路 

分 类 号：R743.33[医药卫生—神经病学与精神病学]