基于组学分析甲磺酸萘莫司他通过调控HMGB1对大鼠肾缺血再灌注模型的保护作用及机制研究  

Nafamostat Mesylate Alleviates Renal Ischemia-Reperfusion Injury in a Rat Model Through HMGB1 Modulation:An Omics Analysis-Based Study of the Protective Effect and the Mechanisms Involved

作　　者：吴煜璇 李泓倩 刘琳丰 王余 陈波 刘江 秦建华 欧三桃 吴蔚桦 WU Yuxuan;LI Hongqian;LIU Linfeng;WANG Yu;CHEN Bo;LIU Jiang;QIN Jianhua;OU Santao;WU Weihua(Department of Nephrology,Affiliated Hospital of Southwest Medical University/Sichuan Provincical Clinical Research Center for Nephrology/Key Laboratory of Metabolic Diseases,Luzhou 646000,China;Department of Nephrology,Dujiangyan People's Hospital,Chengdu 611830,China;Anatomy Teaching and Research Office,School of Basic Medical Sciences,Southwest Medical University,Luzhou 646000,China)

机构地区：[1]西南医科大学附属医院肾内科/四川省肾脏病临床研究中心/四川省代谢性疾病重点实验室,泸州646000 [2]都江堰市人民医院肾脏内科,成都611830 [3]西南医科大学基础医学院解剖教研室,泸州646000

出　　处：《四川大学学报(医学版)》2025年第1期175-182,共8页Journal of Sichuan University(Medical Sciences)

基　　金：泸州市科技人才局社发基金(No.2022-SYF-59)资助。

摘　　要：目的探讨甲磺酸奈莫司他(nafamostat mesylate,NM)对肾缺血再灌注(renal ischemia-reperfusion injury,RIRI)模型的器官保护作用及机制。方法21只健康雄性SD大鼠,随机为3组(n=7),分别为假手术组(Sham组)、肾脏缺血再灌注组(RIRI组)、甲磺酸萘莫司他干预组(NM组)。RIRI组及NM组制作IRI动物模型。NM组造模前给予腹腔注射NM 0.75 mg/kg,造模24 h后收集大鼠静脉血和肾组织标本。血清标本检测血清肌酐、胱抑素C、血清炎症因子水平。肾组织行HE染色、TUNEL染色评估肾组织损伤情况。分别采用免疫荧光及免疫印迹进行HMGB1的定位表达。通过单细胞RNA核测序获得RIRI组与NM组的大鼠肾脏单细胞转录组获得RIRI细胞图谱。根据细胞标志性基因对细胞进行注释,探索疾病模型的细胞类型构成占比、组间免疫细胞的功能状态。结果①RIRI组及NM组胱抑素C和肌酐、炎症因子水平高于Sham组,NM组表达水平低于RIRI组(P<0.05)。NM组肾小管损伤积分、凋亡率低于RIRI组(P<0.05),但两组均高于Sham组。与RIRI组比较,NM组HMGB1表达下降(P<0.05),但与Sham组相比两组表达均升高。免疫荧光显示NM、RIRI组均出现更多的HMGB1细胞质表达,以RIRI组更为明显。②通过单细胞核测序结果得到13大类细胞群体,NM组小管细胞比例更高,其中HMGB1基因在损伤的近曲小管细胞高表达。NM组极化状态的Macro3细胞亚群所占总体巨噬细胞比例均较RIRI组减少。结论NM可能对大鼠RIRI模型具有保护作用,其机制可能与经过调节HMGB1介导的巨噬细胞功能异常有关。Objective To investigate the organ protective role and the underlying mechanism of nafamostat mesylate(NM)in a renal ischemia-reperfusion injury(RIRI)model.Methods A total of 21 healthy male Sprague-Dawley(SD)rats were randomly assigned to 3 groups(n=7 in each group),including the sham operation group(Sham group),the RIRI group,and the NM intervention group(NM group).The RIRI and NM groups underwent ischemiareperfusion injury(IRI)modeling.The NM group was given an intraperitoneal injection of NM at 0.75 mg/kg before modeling.Venous blood and renal tissue samples were then collected from the rats 24 hours after modeling.The levels of serum creatinine,cystatin C,and serum inflammatory factors were determined using the serum samples.Hematoxylineosin(HE)staining and TUNEL stainings were performed on the renal tissues to evaluate the damage of the renal tissues.The localization and expression of HMGB1 were analyzed by immunofluorescence and Western blotting,respectively.Single-cell RNA sequencing of the nuclei was performed to obtain the single-cell transcriptome of the kidneys from the rats in the RIRI and the NM groups and to acquire the RIRI cell profile.The cells were annotated according to the cell marker genes to explore the cell type composition in the disease model and the functional status of immune cells between the groups.Results 1)Compared with those of the Sham group,the levels of cystatin C,creatinine,and inflammatory factors in the RIRI and NM groups were significantly increased,and the expression levels in the NM group were lower than those in the RIRI group(P<0.05).Compared with those of the RIRI group,the tubular injury score and apoptosis rate in the NM group were significantly decreased(P<0.05),but those of both the NM and RIRI groups were higher than those of the sham group.Compared with that in the RIRI group,the expression of HMGB1 in the NM group was significantly decreased(P<0.05),but the expression levels in both the RIRI and NM groups were higher than that in the sham group.Immunofluorescenc

关 键 词：缺血再灌注 急性肾损伤 甲磺酸萘莫司他 高迁移率族蛋白B1 单核巨噬细胞 

分 类 号：R965[医药卫生—药理学]