PPAR-γ/Nrf2介导Heme-Hx-LRP1通路在脑出血后血肿清除中的作用机制  

The mechanism of hematoma clearance followed intracerebral hemorrhage via PPAR-γ/Nrf2 mediated Hxheme-LRP1 pathway

作　　者：董梁 王改青[1,2] 段淑娜 柏琴琴 刘丽荣 Dong Liang;Wang Gaiqing;Duan Shuna;Bai Qinqin;Liu Lirong(Department of Neurology,the Second Clinical Medical College of Shanxi Medical University,Taiyuan 030001,China;Department of Neurology,Sanya Central Hospital(Hainan Third People's Hospital),Sanya 572000,China)

机构地区：[1]山西医科大学第二临床医学院神经内科,太原030001 [2]三亚中心医院(海南省第三人民医院)神经内科,三亚572000

出　　处：《中华脑血管病杂志（电子版）》2021年第6期373-381,共9页Chinese Journal of Cerebrovascular Diseases(Electronic Edition)

基　　金：国家自然科学基金面上项目(81771294);深圳市科创委资助项目(JCYJ20190808161013492);海南省临床医学中心建设项目资助。

摘　　要：目的探讨血红素(Heme)-血红素结合蛋白(Hx)-低密度脂蛋白受体相关蛋白1(LRP1)通路在脑出血后清除血肿中的作用,以及过氧化物酶体增殖物激活受体γ(PPAR-γ)/核因子E2相关因子2(Nrf2)对该通路的调控机制。方法采用Ⅳ型胶原酶大鼠脑出血模型;SD大鼠86只,随机分为手术对照组(n=20)、脑出血组(n=22)、PPAR-γ/Nrf2激动剂(红曲素)+脑出血组(n=21)、PPAR-γ抑制剂(格列卫)+脑出血组(n=23)。术后3 d应用头颅MRI不同序列测定血肿及水肿体积,同期测定血肿周围脑水含量及血红蛋白含量,应用免疫荧光双标法观察LRP1及Hx的表达部位,免疫印迹法测定PPAR-γ、Nrf2、LRP1、Hx的表达量。多组间血肿、水肿体积、血红蛋白含量、脑水含量、PPAR-γ、Nrf2、LRP1、Hx蛋白量比较采用单因素方差分析,组间多重比较采用Tukey's检验;血肿与血红蛋白含量、水肿与脑水含量相关关系采用Pearson相关分析。结果血肿、水肿体积、脑组织脑水含量及血红蛋白含量结果:格列卫+脑出血组>脑出血组>红曲素+脑出血组>手术对照组;LRP1及Hx的表达量:红曲素+脑出血组>脑出血组>格列卫+脑出血组>手术对照组;红曲素+脑出血组的LRP1及Hx在小胶质细胞的表达显著增高,且以血管周围更明显。相关分析显示MRI测量的血肿与脑组织血红蛋白含量呈显著正相关(r=0.956,P=0.005);头颅MRI测量的水肿体积与脑水含量呈显著正相关(r=0.924,P=0.010)。结论红曲素作为PPAR-γ/Nrf2双靶点激动剂,可能通过强化血管周围小胶质细胞Hx及LRP1的表达促进血肿清除,同时减轻脑出血后水肿形成。提示PPAR-γ/Nrf2通过调控Heme-Hx-LRP1通路参与脑出血后血肿清除;头颅MRI测定脑组织血肿及水肿与脑组织检测血红蛋白及脑水含量所得出的结论一致。Objective To explore the role of heme-hemopexin-low density lipoprotein receptor related protein 1(LRP1)pathway in hematoma clearance followed intracerebral hemorrhage(ICH)as well as its regulatory mechanism of peroxisome proliferator-activated receptorγ(PPAR-γ)/nuclear factorerythroid 2 related factor-2(Nrf2).Methods ICH was induced by stereotactic,intrastriatal injection of typeⅣcollagenase.86 male SD rats were randomly divided into sham,ICH+vehicle(normal saline),ICH+monascin(dual PPAR-γ/Nrf2 agonist,10 mg/kg,twice daily),ICH+gleevec(PPAR-γantagonist,100 mg/kg,once a day).Monascin and gleevec were administered by gastric perfusion for 3 days after ICH induction.Hematoma volume and brain edema were assayed by brain magnetic resonance imaging(MRI),and hemoglobin levels and brain water content were evaluated at 3 days after ICH.Brain expression of Hx/LRP1 with microglia were analyzed by double-labelling immunofluorescence and western blot.One-way analysis of variance(ANOVA)was performed for comparisons among multiple groups,and Tukey's test was used for pairwise comparison between groups.The correlation between hematoma and hemoglobin content,edema and cerebral water content were analyzed by Pearson correlation.Results Hematoma volume and brain edema assessed by MRI as well as hemoglobin levels and brain water content showed which were reduced in monascin-treated ICH while increased in gleevec-treated ICH;Expression of LRP1 and Hx on microglia were significantly increased after ICH,especially expressed around vessels in monascin-treated ICH.Pearson correlation analysis revealed that there was a positive correlation between edema volume shown by MRI and brain water content(r=0.956,P=0.005),hematoma volume as shown by MRI and brain hemoglobin levels(r=0.924,P=0.010).Conclusion As a PPAR-γ/Nrf2 dual-target agonist,monascin promoted hematoma removal through inducing LRP1 and Hx expressed on microglia around vessels,while alleviating secondary edema formation followed ICH;It showed that PPAR-γ/Nrf2 participate

关 键 词：脑出血 血肿清除 血红素 血红素结合蛋白 低密度脂蛋白受体相关蛋白1 

分 类 号：R651.1[医药卫生—外科学]