橙皮苷联合丰富环境对脑出血模型小鼠神经保护作用及Nrf2/Gpx4介导的铁死亡机制  

Neuroprotective effect of hesperidin combined with enriched environment on intracerebral hemorrhage model mice and ferroptosis mechanism mediated by Nrf2/Gpx4

作　　者：马瑞 尤红[2] 高雅雅 刘红 包娟 卢苇 马鑫宇 张敏[2] Ma Rui;You Hong;Gao Yaya;Liu Hong;Bao Juan;Lu Wei;Ma Xinyu;Zhang Min(The First Clinical Medical College of Gansu University of Chinese Medicine,Lanzhou 730000,China;The Second Ward of Neurology of Gansu Provincial Hospital,Lanzhou 730099,China;Clinical Medical College of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]甘肃中医药大学第一临床医学院,兰州730000 [2]甘肃省人民医院神经内科二病区,兰州730099 [3]宁夏医科大学临床医学院,银川750004

出　　处：《中华行为医学与脑科学杂志》2024年第11期1013-1019,共7页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：甘肃省自然科学基金(21JR7RA597,24JRRA1052);甘肃省人民医院院内科研基金(22GSSYB-9,22GSSYD-65,22GSSYD-66)。

摘　　要：目的探讨橙皮苷联合丰富环境对胶原酶诱导的脑出血(intracerebral hemorrhage,ICH)模型小鼠的神经保护作用及铁死亡相关作用机制。方法将90只C57BL/6J小鼠按随机数字表分为假手术组、脑出血组、丰富环境组、橙皮苷组、丰富环境+橙皮苷组(联合干预组)、联合干预+Nrf2特异性抑制剂ML385(抑制剂组)共6组,每组15只。抑制剂组、脑出血组、丰富环境组、橙皮苷组、联合干预组小鼠尾壳核内注射0.075 U/μLⅦ型胶原酶溶液0.5μL(用0.9%NaCl溶液溶解)建立ICH模型,假手术组小鼠注射等体积0.9%NaCl溶液。橙皮苷组、联合干预组、抑制剂组小鼠给予橙皮苷溶液(0.5%羧甲基纤维素钠溶解)灌胃,假手术组、脑出血组、丰富环境组小鼠给予等体积0.5%羧甲基纤维素钠溶液灌胃。抑制剂组腹腔注射ML385(30 mg/kg,5%DMSO溶解),其余各组均腹腔注射等体积5%DMSO溶液。灌胃、腹腔注射均在造模手术结束后6 h内完成,1次/d,共14 d。丰富环境组、联合干预组、抑制剂组小鼠在EE鼠笼饲养,假手术组、脑出血组、橙皮苷组小鼠在普通鼠笼饲养。以上干预结束后,所有小鼠均进行神经功能缺损评分(modified neurological severity score,mNSS评分)。并分别进行脑含水量检测、普鲁士蓝染色,ELISA法检测丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、活性氧(reactive oxygen species,ROS)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,Gpx4),PCR和Western blot分别检测核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)、Gpx4 mRNA、蛋白水平的表达。使用GraphPad Prism 9软件对数据进行统计分析,多组间比较采用单因素方差分析,组间多重比较采用Tukey检验。结果(1)6组小鼠mNSS评分差异有统计学意义(F=66.35,P<0.001)。脑出血组[(8.00±1.46)分]mNSS评分高于假手术组[(0.86±0.83)分](P<0.05);丰富环境组[(6.47±1.13)分]、橙皮苷组[(6.13±1.25)分]mNSS评�ObjectiveTo investigate the neuroprotective effect of hesperidin combined with enriched environment on ferroptosis in collagenase-induced intracerebral hemorrhage(ICH)mouse model as well as the ferroptosis mechanism.MethodsICH model was established by injecting collagenaseⅦinto caudate putamen nucleus.Ninty C57BL/6J mice were randomly divided into 6 groups according to a random number table:sham group,intracerebral hemorrhage group,enriched environment group,hesperidin group,enriched environment and hesperidin group(combination group),and combination group+Nrf2 specific inhibitor ML385(inhibitor group),with 15 mice in each group.The mice in inhibitor group,intracerebral hemorrhage group,enriched environment group,hesperidin group and combination group were injected with 0.5μL collagenase typeⅦsolution(0.075 U/μL,dissolved with 0.9%NaCl solutin)for ICH modeling,and the mice in sham group were injected with 0.9%normal saline.The hesperidin group,combination group,and inhibitor group were given hesperidin solution(dissolved in 0.5%carboxymethyl cellulose sodium)by gavage within 6 hours after the modeling surgery.The sham group,intracerebral hemorrhage group,and enriched environment group were given equal volumes of 0.5%carboxymethyl cellulose sodium solution by gavage.The inhibitor group was intraperitoneally injected with Nrf2 specific inhibitor ML385(30 mg/kg,dissolved in 5%DMSO),while the other groups were intraperitoneally injected with an equal volume of 5%DMSO.Both gastric perfusion and intraperitoneal injection were completed within 6 hours after the end of modeling operation,once a day for 14 days.After the postoperative recovery of the mice,the enriched environment group,combination group,and inhibitor group were placed in enriched environment cages,while the sham group,intracerebral hemorrhage group,and hesperidin group were placed in regular cages.After all intervention were completed,all mice were evaluated using the modified neurological severity score(mNSS).Then the mice were subjected to brain w

关 键 词：橙皮苷 脑出血 丰富环境 铁死亡 Nrf2/Gpx4通路 小鼠 

分 类 号：R743.34[医药卫生—神经病学与精神病学]