温阳复元方对脑缺血再灌注损伤大鼠线粒体铁死亡相关蛋白IRP1和FAM96A表达的影响  

Effects of Decoction for warming Yang and recovering vitality on the expression of mitochondrial ferroptosis-related proteins IRP1 and FAM96A in rats with cerebral ischemia-reperfusion injury

作　　者：李丽琴 莫雪妮[1] 袁莉 陈炜[2] 秦红玲[2] 胡跃强[2] 姚春[1] LI Liqin;MO Xueni;YUAN Li;CHEN Wei;QIN Hongling;HU Yueqiang;YAO Chun(Guangxi University of Traditional Chinese Medicine,Nanning 530012,Guangxi,China;The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530020,Guangxi,China)

机构地区：[1]广西中医药大学,广西南宁530012 [2]广西中医药大学第一附属医院,广西南宁530020

出　　处：《现代中西医结合杂志》2023年第15期2045-2050,2059,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金项目(81973768);广西自然科学基金重点项目(2019GXNSFDA245006);广西中医脑病临床研究中心项目(桂科AD20238028);广西研究生教育创新计划项目(YCSZ2022015)。

摘　　要：目的观察温阳复元方对脑缺血再灌注后大鼠铁调节蛋白1(IRP1)和96序列相似家庭成员A(FAM96A)表达的影响,探讨其脑保护机制。方法将84只SD大鼠随机分为假手术组、脑缺血再灌注组、温阳复元方组和补阳还五汤组,每组21只。除假手术组外,其余组采用改良Longa线栓法建立脑缺血再灌注模型。温阳复元方组和补阳还五汤组大鼠在造模前30 min分别给予温阳复元方14.04 g/kg和补阳还五汤11.43 g/kg灌胃,大鼠清醒后1 h继续予以原方灌胃,假手术组、脑缺血再灌注组给予生理盐水灌胃。分别于再灌注12 h、24 h、3 d时对各组大鼠进行神经功能缺损评分,随后处死大鼠取脑组织,分别采用RT-qPCR、免疫组化法检测缺血灶脑组织中IRP1、FAM96AmRNA和蛋白表达情况。结果脑缺血再灌注组大鼠再灌注12 h、24 h、3 d时神经功能缺损评分均明显高于假手术组(P均<0.05);温阳复元方组大鼠再灌注12 h、24 h、3 d时神经功能缺损评分均明显低于同期脑缺血再灌注组(P均<0.05),且再灌注3 d时明显低于同期补阳还五汤组(P<0.05);补阳还五汤组仅在再灌注12 h时神经功能缺损评分明显低于脑缺血再灌注组(P<0.05)。脑缺血再灌注组再灌注12 h、24 h、3 d时IRP1、FAM96A mRNA相对表达量和蛋白表达平均光密度值均明显高于同期假手术组(P均<0.05);温阳复元方组IRP1、FAM96A mRNA相对表达量和蛋白表达平均光密度值随再灌注时间延长逐渐降低,再灌注3 d时最低,与脑缺血再灌注组相应时间点比较差异均有统计学意义(P均<0.05),且再灌注3 d时IRP1 mRNA相对表达量和再灌注24 h、3 d时FAM96A mRNA相对表达量及IRP1、FAM96A蛋白表达平均光密度值均明显低于同期补阳还五汤组(P均<0.05);补阳还五汤组IRP1、FAM96A mRNA相对表达量和蛋白表达平均光密度值随再灌注时间延长逐渐升高,但再灌注12 h、24 h时均明显低于同期脑缺血再灌注组(P均<0.05),再灌�Objective It is to observe the effects of Decoction for warming Yang and recovering vitality(DWYRV)on the expression of iron-regulated protein 1(IRP1)and 96 sequence similar family member A(FAM96A)in rats after cerebral ischemia and reperfusion,and to explore its cerebral protection mechanism.Methods Eighty-four SD rats were randomly divided into sham operation group,cerebral ischemia-reperfusion group,DWYRV group and Decoction for invigorating Yang for recuperation(DIYR)group,with 21 rats in each group.The cerebral ischemia/reperfusion models were established using the modified Longa line bolus method in all groups except the sham operation group.The rats in the DWYRV group and DIYR group were gavaged with 14.04 g/kg of DWYRV and 11.43 g/kg of DIYR,respectively,30 min before the modeling,and were continuously gavaged with the original decoction for 1 h after they woke up,and the rats in the sham operation group and cerebral ischemia reperfusion group were gavaged with physiological saline.The neurological deficits of the rats in each group were scored at 12 h,24 h and 3 d after reperfusion,and then the brain tissues were taken from the executed rats,to detect the expressions of IRP1,FAM96A mRNA and protein in the brain tissues of ischemic foci by RT-qPCR and immunohistochemistry,respectively.Results The neurological deficit scores of rats in the cerebral ischemiareperfusion group were significantly higher than those of the sham operation group at 12 h,24 h and 3 d after reperfusion(all P<0.05);the neurological deficit scores of rats in the DWYRV group at 12 h,24 h and 3 d after reperfusion were significantly lower than those of the cerebral ischemia-reperfusion group at the same period(all P<0.05),and the scores at 3 d after reperfusion were significantly lower than those of the DIYR group at the same period(P<0.05);the neurological deficit scores were significantly lower than those in the cerebral ischemia-reperfusion group only at 12 h after reperfusion(P<0.05).The relative expression of IRP1,FAM96A mRNA and t

关 键 词：脑缺血再灌注损伤 线粒体铁死亡 温阳复元方 铁调节蛋白1 96序列相似家庭成员A 

分 类 号：R-332[医药卫生]