异泽兰黄素对脑出血继发性脑损伤的神经保护作用及相关机制研究  

Neuroprotective effect and related mechanisms of of eupatilin on secondary brain injury after intracerebral hemorrhage

作　　者：申友奎[1] 刘飞飞 王雁秋 SHEN Youkui;LIU Feifei;WANG Yanqiu(Department of Neurosurgery,Hangzhou Hospital of Traditional Chinese Medicine,Hangzhou 310007,China;不详)

机构地区：[1]杭州市中医院神经外科,310007 [2]杭州市中医院内分泌科,310007

出　　处：《心电与循环》2024年第4期323-331,342,I0002,共11页Journal of Electrocardiology and Circulation

基　　金：浙江省中医药科技计划项目(2021ZA105、2023ZL545);杭州市医药卫生科技项目(A20200040)。

摘　　要：目的 探讨异泽兰黄素(EP)对脑出血(ICH)继发性脑损伤的神经保护作用及相关机制。方法 体内动物实验:将无特定病原体雄性小鼠按随机数字表法分为假手术组(0.9%氯化钠溶液)、模型组(0.9%氯化钠溶液)、EP低剂量组(5 mg/kg EP)、EP中剂量组(10 mg/kg EP)、EP高剂量组(20 mg/kg EP),每组18只;除假手术组外,其余4组均采用Ⅳ型胶原酶诱导ICH法构建小鼠ICH模型。检测并比较5组小鼠给药7 d后改良神经功能缺失评分(m NSS)、水迷宫实验结果(逃避潜伏期、平台停留时间、穿越平台次数)、血脑屏障通透性、脑含水量、脑组织病理学改变、氧化应激指标[丙二醛(MDA)、超氧物歧化酶(SOD)、过氧化氢酶(CAT)]、细胞凋亡数、炎症因子指标[肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、白细胞介素1β(IL-1β)]以及凋亡相关蛋白[B淋巴细胞瘤-2(Bcl-2)、Bcl-2关联X蛋白(Bax)、裂解的胱天蛋白酶-3(Cleaved caspase-3)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)、糖原合成酶-3β(GSK-3β)、磷酸化PI3K(p-PI3K)、磷酸化Akt(p-Akt)、磷酸化GSK-3β(p-GSK-3β)]、水通道蛋白4(AQP4)蛋白表达水平。体外细胞实验:将培养后的人神经母细胞瘤细胞系SH-SY5Y细胞分别设空白组、模型组、EP低剂量组(12.5μmol/L)、EP中剂量组(25μmol/L)、EP高剂量组(50μmol/L),每组设6复孔;除空白组不作任何处理外,其余4组均加入冈田酸构建ICH神经细胞损伤模型。检测并比较4组细胞存活率、细胞凋亡率以及凋亡相关蛋白表达水平。结果 体内动物实验结果显示,与假手术组比较,模型组小鼠脑组织结构破坏明显,神经细胞坏死且大量炎性细胞浸润,m NSS评分、逃避潜伏期、血脑屏障通透性、脑含水量以及脑组织MDA、细胞凋亡数和Cleaved caspase-3、AQP4蛋白表达水平均明显升高(均P<0.05),平台停留时间、穿越平台次数、体重以及脑组织SOD、CAT、Bcl-2/Bax、p-PI3K/PI3K、p-AMPK/AMPObjective To explore the neuroprotective effect of eupatilin(EP) on secondary brain injury after intracerebral hemorrhage(ICH) and the underlying mechanisms. Methods In vivo animal experiments, male mice were randomly divided into five groups using the random number table method. The sham surgery group(0.9% sodium chloride solution), the model group(0.9% sodium chloride solution), the EP low-dose group(5 mg/kg EP), the EP medium dose group(10 mg/kg EP) and the EP high-dose group(20 mg/kg EP), 18 mice in each group. Except for the sham surgery group, the other 4 groups underwent ICH model construction using type Ⅳ collagenase. After 7 days of medication, we evaluated and compared the modified neurologic severity score(m NSS), water maze test results(escape latency, platform residence time, number of platform crossings), blood-brain barrier permeability, brain water content,pathological changes in brain tissue, oxidative stress indicators [malondialdehyde(MDA), superoxide dismutase(SOD),catalase(CAT)], cell apoptosis, inflammatory factor indicators [tumor necrosis factor alpha(TNF-α), interleukin-6(IL-6),interleukin-1 β(IL-1β)] and apoptosis related proteins [B-cell lymphoma-2(Bcl-2), Bcl-2 associated X protein(Bax),Cleaved caspase-3, phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), glycogen synthetase-3(GSK-3),phosphorylated PI3K(p-PI3K), phosphorylated Akt(p-Akt), phosphorylated GSK-3β(p-GSK-3β)] and aquaporin 4(AQP4) in 5 groups. In vitro cell experiments, the cultured SH-SY5Y cells were divided into the control group, the model group, the EP low-dose group(12.5 μmol/L), the EP medium dose group(25 μmol/L) and the EP high-dose group(50μmol/L), 6 wells in each group. Except for the control group, the other 4 groups were added with okada acid to construct an ICH neural cell injury model. The cell survival rate, apoptosis rate and expression levels of apoptosis related proteins in 4 groups were detected. Results The results of vivo experiment showed that compared with the sham surgery group, th

关 键 词：异泽兰黄素 脑出血 继发性脑损伤 磷脂酰肌醇3-激酶/蛋白激酶B/糖原合成酶-3β信号通路 血脑屏障 

分 类 号：R285.5[医药卫生—中药学]