基于Nrf2/HO-1通路探讨益气活血化浊解毒方改善大鼠脑缺血再灌注损伤的实验研究  

Experimental Study of Effect of Yiqi Huoxue Huazhuo Jiedu Formula on Cerebral Ischemia-reperfusion Injury in Rats Based on Nrf2/HO-1 Pathway

作　　者：石瑞 孙亚萍 谢占维 孙玲玲 崔怡娴 田军彪 SHI Rui;SUN Ya-ping;XIE Zhan-wei;SUN Ling-ling;CUI Yi-xian;TIAN Jun-biao(Cerebral Ischemia-reperfusion Injury in Rats Based on Nrf2/HO-1 Pathway,Shijiazhuang 050091)

机构地区：[1]河北中医学院研究生学院,石家庄050091 [2]河北中医学院第一附属医院,石家庄050011

出　　处：《河北中医药学报》2023年第1期3-8,23,共7页Journal of Hebei Traditional Chinese Medicine and Pharmacology

基　　金：河北省省级科技计划资助(重点研发计划)项目:No.20377710D;河北省中医药管理局科研计划重点项目:No.Z2022008;河北中医学院科研能力提升重点项目:No.KTZ2019028;河北中医学院科研能力提升一般项目:No.KTZ2019013;河北中医学院研究生创新资助项目:No.XCXZZBS2022008

摘　　要：目的:研究益气活血化浊解毒方对大鼠脑缺血再灌注损伤(CIRI)的脑保护作用,及其与核因子E2相关因子2/血红素加氧酶-1(Nrf2/HO-1)信号通路活性的相关性。方法:72只SD大鼠随机分为6组(n=12):假手术组、模型组、尼莫地平组和益气活血化浊解毒方高、中、低剂量组。除假手术组外均采用线栓法建立大鼠大脑中动脉闭塞再灌注(MCAO/R)模型。再灌注后假手术组、模型组大鼠给予生理盐水,尼莫地平组和益气活血化浊解毒方各剂量组给予相应剂量药液。连续灌胃3 d后,对比各组间大鼠神经功能缺损评分;2,3,5-氯化三苯基四氮唑(TTC)染色检测脑梗死体积;原位末端标记法(TUNEL)检测脑组织细胞凋亡情况;Western blot检测脑组织Nrf2和HO-1蛋白的表达情况,实时荧光定量PCR(qRT-PCR)检测脑组织Nrf2和HO-1 mRNA的表达情况;试剂盒检测脑组织中超氧化物歧化酶(SOD)活性、丙二醛(MDA)和一氧化氮(NO)含量。结果:与假手术组比较,模型组大鼠神经功能缺损评分与脑梗死体积显著增高(P<0.01),大鼠脑组织凋亡细胞显著增多(P<0.01),Nrf2、HO-1蛋白及Nrf2、HO-1 mRNA表达水平均显著下降(P<0.01),SOD活性显著降低、MDA和NO含量显著增高(P<0.01)。与模型组比较,益气活血化浊解毒方各剂量组大鼠神经功能缺损评分与脑梗死体积明显降低(P<0.05),大鼠脑组织凋亡细胞显著减少(P<0.01),Nrf2、HO-1蛋白表达水平明显升高(P<0.05),Nrf2、HO-1 mRNA表达水平显著升高(P<0.01),SOD活性明显增高、MDA含量明显降低、NO含量显著降低(P<0.05,P<0.05,P<0.01)。结论:益气活血化浊解毒方可能通过激活Nrf2/HO-1信号通路发挥抗氧化应激作用,减少神经细胞损伤,进而达到脑保护的作用。Objective:To study the neuroprotective effect of Yiqi Huoxue Huazhuo Jiedu Formula(YHJF)on cerebral ischemia reperfusion injury(CIRI)in rats and its correlation with the activity of nuclear factor E2 related factor 2/heme oxygenase-1(Nrf2/HO-1)signal pathway.Methods:72 SD rats were randomly divided into 6 groups(n=12):sham operation group,model group,nimodipine group,high,medium and low dose groups of YHJF.Except the sham operation group,the middle cerebral artery occlusion and reperfusion(MCAO/R)model of rats was established by thread embolism method.Then,the sham operation group and the model group were given physiological saline,and the nimodipine group and each dose group of YHJF were given corresponding doses of liquid medicine.After continuous gavage for 3 days,the scores of nerve function defect of rats in each group were compared;The volume of cerebral infarction was measured by 2,3,5-triphenyl tetrazolium chloride(TTC)staining;The apoptosis of brain tissue was detected by TUNEL;Western blot was used to detect the expression of Nrf2 and HO-1 protein and real-time fluorescent quantitative PCR(qRT-PCR)to detect the expression of Nrf2 and HO-1 mRNA in brain tissue;The activity of superoxide dismutase(SOD),the content of malondialdehyde(MDA)and nitric oxide(NO)were detected with the kit.Results:Compared with the sham operation group,the score of neural function defect and the volume of cerebral infarction in the model group were significantly increased(P<0.01),the number of apoptotic cells in the brain tissue was significantly increased(P<0.01),the expression levels of Nrf2,HO-1 protein and Nrf2,HO-1 mRNA were significantly decreased(P<0.01),the activity of SOD was significantly decreased,and the content of MDA and NO was significantly increased(P<0.01).Compared with the model group,the score of nerve function defect and the volume of cerebral infarction in each dose group of YHJF were significantly decreased(P<0.05),the apoptotic cells in brain tissue were significantly decreased(P<0.01),the expression level

关 键 词：益气活血化浊解毒方 脑缺血再灌注损伤 中风 缺血性脑卒中 核因子E2相关因子2 血红素加氧酶-1 氧化应激 

分 类 号：R285.5[医药卫生—中药学]