黄芪甲苷对缺血性卒中小鼠神经功能保护作用及机制研究_毕芳芳.pdf

89现代检验医学杂志第 38 卷第 1 期2023 年 1 月J Mod Lab Med,Vol.38,No.1,Jan.2023黄芪甲苷对缺血性卒中小鼠神经功能保护作用及机制研究毕芳芳，史敏，程微微，张露（西安培华学院医学检验技术系，西安710199）摘要：目的探讨黄芪甲苷（astragaloside-IV,AST-IV）对缺血性卒中（ischemic stroke）模型鼠神经功能保护作用及机制研究。方法选取改良线栓法制备大脑中动脉梗死（middle cerebral artery occlusion，MCAO）小鼠模型，小鼠分成四组，每组 8 只，分别为假手术组（Sham）、药物对照组（AST-IV）、MCAO 组和治疗组（AST-IV+MCAO）；采用改良神经功能损伤评分（modified neurological severity score，mNSS）实验和转棒（Rotarod）实验综合评价 AST-IV 的神经保护功能；采用 ELISA 法检测 AST-IV 对 MCAO 脑组织核转录因子(NF-KB)、Toll 样受体 4(TLR-4)、肿瘤坏死因子(TNF-)、胶质纤维酸性蛋白(glial fibrillary acidic protein，GFAP)、血红素氧合酶-1(HO-1)以及半胱天冬酶-1(caspase-1)表达水平的影响；采用比色法检测 AST-IV 对 MCAO 脑组织丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)和活性氧（ROS）表达水平的影响。结果MCAO组的mNSS评分（54.068.52）、脑组织ROS（54.068.52 AFU）、MDA（2.400.33 nmol/mg），NF-B（304.9439.93pg/mg），TLR4（225.6037.51pg/mg），TNF-（80.1411.06pg/mg），GFAP（181.2419.88pg/mg）和 caspase-1（1.760.20）表达均显著高于 Sham 组（0.750.89，19.852.45AFU，1.430.29 nmol/mg，131.0329.45 pg/mg，102.7419.18 pg/mg，32.0410.30 pg/mg，63.1414.48 pg/mg，1.000.09)，差异具有统计学意义（F=38.572 184.478，均 P0.001）；MCAO 组在 Rotarod 上的停留时间（74.6318.73s），脑组织 SOD（2.880.49unin/mg），GSH（3.490.64nmol/mg）和 HO-1（105.9324.45pg/ml）表达均显著低于 Sham 组（186.5048.10s，5.580.71unit/mg，6.320.82 pg/ml，203.1128.65pg/ml），差异具有统计学意义（F=37.58678.889，均P0.001）；AST-IV+MCAO组的mNSS评分（3.881.36）、脑组织ROS（24.6414.26AFU），MDA（2.000.21nmol/mg），NF-B（212.0338.63 pg/mg），TLR-4（134.8125.60 pg/mg），TNF-（53.6112.49 pg/mg），GFAP（101.0015.04 pg/mg）和 caspase-1（1.460.15）表达均显著低于 MCAO 组，差异具有统计学意义（F=4.639 82.890，均 P0.05）；AST-IV+MCAO 组在 Rotarod 上的停留时间（160.7533.33s），脑组织 SOD（4.510.51unit/mg），GSH（5.300.73nmol/mg）和 HO-1（179.9620.97pg/ml）均显著高于 MCAO 组，差异具有统计学意义（F=27.681 42.364，均 P0.001）。结论AST-IV 的脑保护作用可能与其抗炎、抗氧化应激、抑制焦亡进程有关。关键词：黄芪甲苷；缺血性卒中；炎症；氧化应激；焦亡中图分类号：R-332文献标识码：A文章编号:1671-7414（2023）01-089-05doi:10.3969/j.issn.1671-7414.2023.01.017Study on the Protective Effect and Mechanism of Astragaloside IV on Neurological Function in Mice with Ischemic StrokeBI Fang-fang,SHI Min,CHENG Wei-wei,ZHANG Lu（Department of Medical Laboratory Technology,Xian Peihua University,Xian 710199，China）Abstract:ObjectiveTo investigate the protective effect and mechanism of astragaloside IV（AST-IV）in a mouse model of ischemic stroke.MethodsThe middle cerebral artery occlusion(MCAO)mouse model was established by the modified suture method.The mice were divided into four groups with 8 mice in each group,namely the Sham group,the AST-IV group,the MCAO group and the AST-IV+MCAO group.The neuroprotective function of AST-IV was comprehensively evaluated by modified neurological severity score(mNSS)test and Rotarod test.ELISA was used to detect the effect of AST-IV on the expression levels of nuclear factor kappa-B（NF-B）,Toll like receptor 4（TLR-4）,Tumor necrosis factor-（TNF-）,glial fibrillary acidic protein(GFAP),heme oxygenase-1(HO-1)and caspase-1in MCAO brain tissue.Colorimetry was used to detect the effect of AST-IV on the expression levels of malondialdehyde(MDA),Superoxide dismutase（SOD）,glutathione(GSH)and reactive oxygen species（ROS）in MCAO brain tissue.ResultsThe mNSS score（54.068.52）in MCAO group,the expression of ROS（54.068.52 AFU）,MDA（2.400.33 nmol/mg）,NF-B（304.9439.93pg/mg）,基金项目：陕西省教育厅科研计划项目（21JK0820），陕西省自然科学基础研究计划（2022JQ-876，2022JM-558）。作者简介：毕芳芳（1991-），女，硕士研究生，讲师，研究方向：神经炎症，E-mail:。通讯作者：张露（1987-），男，硕士研究生，主管技师，E-mail:。90现代检验医学杂志第 38 卷第 1 期2023 年 1 月J Mod Lab Med,Vol.38,No.1,Jan.2023TLR-4（225.6037.51pg/mg）,TNF-（80.1411.06pg/mg）,GFAP（181.2419.88pg/mg）and caspase-1（1.760.20）in brain tissue were significantly higher than those in the Sham group（0.750.89，19.852.45AFU，1.430.29 nmol/mg，131.0329.45 pg/mg，102.7419.18 pg/mg，32.0410.30 pg/mg，63.1414.48 pg/mg，1.000.09),and the differences were statistically significant(F=8.572 184.478,all P0.001).The MCAO group had a longer residence time（74.6318.73s）on Rotarod,the expression of SOD（2.880.49unin/mg）,GSH（3.490.64nmol/mg）and HO-1（105.9324.45pg/ml）in brain tissue were significantly lower than those in the Sham group（186.5048.10s，5.580.71unit/mg，6.320.82 pg/ml，203.1128.65pg/ml）,and the differences were statistically significant(F=37.58678.889,all P0.001).The mNSS score（3.881.36）in the AST-IV+MCAO group,the expression of ROS（24.6414.26AFU）,MDA（2.000.21nmol/mg）,NF-B（212.0338.63 pg/mg）,TLR-4（134.8125.60 pg/mg）,TNF-（53.6112.49 pg/mg）,GFAP（101.0015.04 pg/mg）and caspase-1（1.460.15）in the brain tissue were significantly lower than MCAO group,and the differences were statistically significant(F=4.639 82.890，all P0.05).AST-IV+MCAO group stay on Rotarod time（160.7533.33s）,the expression of SOD（4.510.51unit/mg）,GSH（5.300.73nmol/mg）and HO-1（179.9620.97pg/ml）were significantly higher than those in the MCAO group,and the differences were statistically significant(F=27.681 42.364,all P0.001).ConclusionThe brain protective effect of AST-IV may be related to its anti inflammation,anti-oxidative stress,and inhibition of coke death process.Keywords:astragaloside IV；ischemic stroke；inflammation；oxidative stress；pyroptosis缺血性脑卒中，也称脑梗死，指由于脑的供血动脉狭窄或闭塞、供血不足导致的脑组织坏死。缺血性脑卒中大约占临床上卒中疾病总数的60%80%，可反复发作，有一定的致残率和致死率