大黄酚对局灶性脑缺血再灌注小鼠缺血半暗带区HIF-1α与VEGF表达的影响  

Effects of chrysophanol on HIF-1αand VEGF expressions in mice with focal cerebral ischemia/reperfusion injury

作　　者：房亚兰 杨楠 赵咏梅 黄语悠 李锦程 段云霞 高利 罗玉敏 Fang Yalan;Yang Nan;Zhao Yongmei;Huang Yuyou;Li Jincheng;Duan Yunxia;Gao Li;Luo Yumin(General Medical Treatment Ward,Pinggu District Hospital of Beijing,Beijing 101200,China;Xuanwu Hospital,Capital Medical University,Beijing Geriatric Medical Research Center,Beijing 100053,China)

机构地区：[1]北京市平谷区医院全科医疗科,北京101200 [2]首都医科大学宣武医院北京市老年病医疗研究中心,北京100053

出　　处：《首都医科大学学报》2021年第2期219-224,共6页Journal of Capital Medical University

基　　金：国家自然科学基金(81971095,81620108011),国家临床重点专科(中医,财社122号)。

摘　　要：目的研究大黄酚(chrysophanol,CHR)对局灶性脑缺血再灌注小鼠缺血半暗带区缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)及血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的影响,探究CHR对脑缺血再灌注损伤的长期脑保护机制。方法采用数字表法随机将18只健康雄性2月龄C57BL小鼠分为3组:假手术(Sham)组、大脑中动脉梗塞(middle cerebral artery occlusion,MCAO)组、CHR组(自造模当日至脑缺血再灌注后14 d按0.1 mg·kg^(-1)·d^(-1)腹腔注射CHR),每组6只。线栓法制作小鼠MCAO模型,缺血45 min后拔出线栓进行再灌注。于再灌注14 d将小鼠处死、取脑,通过免疫荧光染色方法检测脑组织冰冻切片缺血半暗带区内HIF-1α及VEGF水平,并通过免疫荧光双标染色方法确定HIF-1α、VEGF是否在神经元中表达。结果1)Sham组小鼠脑内偶见HIF-1α阳性细胞。MCAO组小鼠脑缺血半暗带区HIF-1α的表达水平比Sham组显著升高(P<0.05)。经CHR治疗14 d的脑缺血再灌注小鼠半暗带区内HIF-1α的表达水平比MCAO组显著减少(P<0.05)。2)Sham组小鼠脑内可见大量的VEGF阳性细胞。MCAO组小鼠脑缺血半暗带区VEGF的表达水平比Sham组显著降低(P<0.05)。经CHR治疗14 d的脑缺血再灌注小鼠半暗带区内VEGF的表达水平比MCAO组显著增加(P<0.05)。3)在小鼠脑缺血半暗带区,HIF-1α或VEGF分别与神经元标志物NeuN共定位。结论CHR可能通过下调HIF-1α的表达水平、上调VEGF的表达水平,减少神经元损伤,从而对脑缺血再灌注损伤发挥长期神经保护作用。Objective To investigate the effects of chrysophanol(CHR)on the expressions of hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)in the penumbra of focal cerebral ischemia/reperfusion mice and further,to explore the long-term protective effect mechanism of CHR against cerebral ischemia/reperfusion injury.Methods According to the random number table,18 healthy male 2-month-old C57BL mice were divided into 3 groups:sham operation(Sham)group(n=6),middle cerebral artery occlusion(MCAO)group(n=6),and CHR group(CHR in dose of 0.1 mg/kg was intraperitoneally injected in CHR group mice once a day for 14 days after reperfusion,n=6).The MCAO for 45 min was induced by thread embolism and the reperfusion lasted for 14 days.On the 14th day after reperfusion,the brain tissue was obtained.The expressions of HIF-1αand VEGF in the brain ischemic penumbra were observed with immunofluorescence labeling.The colocalization of HIF-1αor VEGF with neuron marker NeuN was determined with double labeling immunofluorescence.Results 1)There was little HIF-1αpositive cell in Sham group.Compared with Sham group,the HIF-1αpositive cells in the penumbra of MCAO group obviously increased on the 14th day after reperfusion(P<0.05).Compared with MCAO group,the HIF-1αpositive cells decreased significantly in the penumbra of CHR group on the 14th day after reperfusion(P<0.05).2)There were many VEGF positive cells in Sham group.Compared with Sham group,the VEGF positive cells in the penumbra of MCAO group obviously decreased on the 14th day after reperfusion(P<0.05).Compared with MCAO group,the VEGF positive cells increased significantly in the penumbra of CHR group on the 14th day after reperfusion(P<0.05).3)In the penumbra of ischemic brain of mice,HIF-1αor VEGF was colocalized with neuron marker NeuN,respectively.Conclusion CHR may inhibit the expression of HIF-1αprotein,upregulate the expression of VEGF protein,and reduce neuronal damage,thereby exerting long-term neuroprotection against cerebral ischemia-reper

关 键 词：大黄酚 脑缺血再灌注 缺氧诱导因子-1Α 血管内皮生长因子 

分 类 号：R743.3[医药卫生—神经病学与精神病学]