免疫蛋白酶体抑制剂调控17型辅助性T淋巴细胞/调节性T淋巴细胞免疫平衡在新生大鼠缺氧缺血性脑损伤中的作用  

Immunoproteasome inhibitors regulate the balance of T helper cells 17/regulatory T cells on hypoxic-ischemic brain injury of neonatal rats

作　　者：黄林 周月 苟知贤 胡兴 张峰 鲁利群[1] Huang Lin;Zhou Yue;Gou Zhixian;Hu Xing;Zhang Feng;Lu Liqun(Department of Pediatrics,the First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院第一附属医院儿科,610500

出　　处：《中华实用儿科临床杂志》2020年第23期1821-1824,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：四川省科技厅科研重点项目课题(2018JY0301);成都医学院创新研究团队(CYTD18-06)。

摘　　要：目的探讨免疫蛋白酶体抑制剂PR-957对新生大鼠缺氧缺血性脑损伤(HIBD)的作用及机制。方法将54只7日龄新生大鼠,按随机数字表法分为3组:假手术组、缺氧缺血组(HIBD组)和PR-957治疗组(PR-957组)。按照改良Rice法制备HIBD大鼠模型,PR-957组于造模后腹腔注射PR-957(20 mg/kg),HIBD组于造模后予等体积二甲基亚砜(DMSO)溶液,假手术组不予结扎和低氧处理。造模后72 h,采用HE染色观察大鼠脑皮质炎症情况;免疫组织化学分析左侧脑皮质白细胞介素(IL)-17、IL-10表达;Western blot检测左侧脑组织中(LMP7)、叉头样蛋白3(FOXP3)、维A酸受体相关孤儿受体γt(RORγt)表达;流式细胞术检测外周血17型辅助性T淋巴细胞(Th)、调节性T淋巴细胞(Treg)比例。结果HE染色:假手术组脑组织结构基本正常,HIBD组大鼠左侧脑皮质炎症明显,PR-957组病理改变较HIBD组减轻。免疫组织化学结果:HIBD组大鼠左侧脑皮质IL-10阳性细胞数[(12.11±3.73)%]均显著低于假手术组[(29.12±3.95)%]及PR-957组[(22.61±6.59)%],差异均有统计学意义(均P<0.05);HIBD组左侧脑皮质IL-17阳性细胞数[(35.55±4.85)%]均高于假手术组[(8.48±2.58)%]及PR-957组[(19.16±4.31)%],差异均有统计学意义(均P<0.05)。Western blot结果:HIBD组左侧脑组织LMP7、RORγt蛋白表达(1.01±0.12、0.71±0.10)均高于假手术组(0.50±0.10、0.34±0.07)及PR-957组(0.65±0.13、0.54±0.07),差异均有统计学意义(均P<0.05);HIBD组左侧脑组织FOXP3蛋白表达(0.44±0.10)均低于假手术组(0.93±0.07)及PR-957组(0.68±0.09),差异均有统计学意义(均P<0.05)。外周血流式细胞术:HIBD组Th17/Treg比例(0.66±0.24)均高于假手术组(0.20±0.09)及PR-957组(0.45±0.18),差异均有统计学意义(均P<0.05)。结论PR-957可能通过调控Th17/Treg细胞免疫平衡,减轻HIBD新生大鼠脑组织炎症。Objective To explore the effects and mechanism of PR-957 on hypoxic-ischemic brain damage(HIBD)of newborn rats.Methods A total of 54 rats aged seven days were recruited and randomly divided into sham operation group,HIBD group and PR-957 intraperitoneal group.HIBD model was established according to modified Rice method.PR-957 group was given intraperitoneal injection PR-957(20 mg/kg)after hypoxic-ischemic.Model group was intraperitoneally injected with equal volume of DMSO.Ligation or hypoxic treatment were not given in sham operation group.HE staining was applied to observe inflammation in cortex.Immunohistochemical analysis was adopted to observe the expression of interleukin(IL)-17 and IL-10 in brain tissue.Western blot was carried out to test the protein level of low molecular poly peptide 7(LMP7),forkhead box P3(FOXP3),and retinoic acid-recepter-related orphan receptor gamma t(RORγt).The proportion of T helper cells 17(Th17)/Treg was detected by flow cytometry.Results HE staining displayed that sham operation group brain structure was basically normal,HIBD group revealed significant inflammation in the left cerebral cortex,while some pathological improvement was observed in PR-957 group.Immunohistochemistry:IL-10 positive cells in left cortex of HIBD group[(12.11±3.73)%]were lower than sham operation group[(29.12±3.95)%]and PR-957 group[(22.61±6.59)%],and the differences were statistically significant(all P<0.05).IL-17 positive cells in the left cortex of HIBD group[(35.55±4.85)%]were higher than sham operation group[(8.48±2.58)%]and PR-957 group[(19.16±4.31)%],and the differences were statistically significant(all P<0.05).Western blot:the expression of LMP7 and RORγt in HIBD group(1.01±0.12,0.71±0.10)were higher than those in sham operation group(0.50±0.10,0.34±0.07)and PR-957 group(0.65±0.13,0.54±0.07),and the differences were statistically significant(P<0.05).The expression of FOXP3 in HIBD group(0.44±0.10)was lower than sham operation group(0.93±0.07)and PR-957 group(0.68±0.09),and the d

关 键 词：免疫蛋白酶体 缺氧缺血性脑损伤 新生大鼠 炎症 

分 类 号：R722.1[医药卫生—儿科]