阻断程序性死亡配体-1改善糖尿病脓毒症小鼠免疫状态并对肾脏起保护作用  

The impact of PD-L1 blocking on immune status and renal protection in the models of diabetic mice with sepsis

作　　者：杨黎 高倩 杨铁城 郭树彬[2] Yang Li;Gao Qian;Yang Tiecheng;Guo Shubin(Department of Emergency,Emergency and Critical Care Medical Center,Beijing Shijitan Hospital,Capital Medical University,Beijing 100038,China)

机构地区：[1]首都医科大学附属北京世纪坛医院急诊科急危重症医学中心,北京100038 [2]首都医科大学附属北京朝阳医院急诊医学临床研究中心心肺脑复苏北京市重点实验室,北京100020

出　　处：《中国急救医学》2024年第8期683-689,共7页Chinese Journal of Critical Care Medicine

基　　金：心肺脑复苏北京市重点实验室2019年开放课题(2019XFN-KFKT-02)。

摘　　要：目的探讨阻断程序性死亡配体-1(PD-L1)对糖尿病脓毒症小鼠免疫状态的改变及肾功能保护的可能机制。方法C57BL/6J雄性小鼠随机分为四组:非糖尿病鼠假手术组(Sham组)、糖尿病鼠假手术组(DMS组)、糖尿病鼠脓毒症模型组(DMCLP组)和PD-L1抗体治疗组(anti-PD-L1组),每组6只。采用高糖高脂饮食喂养联合链脲佐菌素(streptozotocin,STZ)注射构建糖尿病小鼠模型,模型建立后继续饲养28 d,盲肠结扎穿孔术(cecal ligation and puncture,CLP)构建脓毒症小鼠模型,PD-L1抗体治疗组于小鼠CLP后1 h腹腔注射抗PD-L1抗体50μg/只。24 h后采用流式细胞术观察各组小鼠经典单核细胞亚型及其表面组织相容性复合体Ⅱ(MHCⅡ)表达,酶联免疫吸附实验(ELISA)检测血清炎症因子IL-6、TNF-α、IL-10表达,全自动生化检测仪检测血清尿素氮(BUN)和肌酐(SCr)水平,苏木素-伊红(HE)染色观察小鼠肾组织损伤情况。结果与Sham组和DMS组比较,DMCLP组小鼠经典单核细胞(Ly6C hi单核细胞)百分比明显增加,其表面MHCⅡ表达明显减少,外周血炎症因子IL-6、TNF-α、IL-10水平升高,BUN和SCr水平明显升高,差异有统计学意义(均P<0.05);肾脏病理损害加重。与DMCLP组比较,anti-PD-L1治疗组小鼠Ly6C hi单核细胞百分比减少,其表面MHCⅡ表达明显增加,外周血炎症因子IL-6、TNF-α、IL-10水平下降,BUN和SCr水平降低,差异有统计学意义(均P<0.05);肾脏病理损害减轻。结论PD-L1抗体通过改善经典单核细胞功能而改变糖尿病脓毒症小鼠免疫状态,进而对肾脏起保护作用。Objective To investigate the mechanism of programmed death-ligand 1(PD-L1)blocking on the change of immune status and the improvement of renal function in diabetic mice with sepsis.Methods C57BL/6J male mice were randomly divided into 4 groups(n=6):non-diabetic mice sham group(Sham group),diabetic mice sham group(DMS group),diabetic mice cecal ligation and puncture(CLP)group(DMCLP group),PD-L1 antibody treatment group(anti-PD-L1 group).The diabetic mouse model was constructed by feeding a high-sugar and high-fat diet combined with streptozotoci(STZ)injection,the feeding after modeling continued for 28 d and CLP were performed for establishing the sepsis mouse model,PD-L1 antibody treatment group mice were intraperitoneally injected with anti-PD-L1 antibody 1 h after CLP(50μg/mouse).Flow cytometry was used to observe the classical monocyte subset and major histocompatibility complexⅡ(MHCⅡ)expression on Ly6C hi monocytes surface in each group.The expressions of inflammatory factors interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and interleukin-10(IL-10)in each group were detected by enzyme-linked immunosorbent assay(ELISA),the blood urea nitrogen(BUN)and serum creatinine(SCr)in mice in each group were detected by automatic biochemical detector,the pathological damage of renal tissue was observed by hematoxylin-eosin(HE)staining.Results Compared with Sham group and DMS group respectively,the percentage of Ly6C hi monocytes in DMCLP group was significantly increased,the expression of MHCⅡon the Ly6C hi monocytes surface was significantly decreased,the levels of inflammatory factors IL-6,TNF-αand IL-10 in peripheral blood were increased,BUN and SCr levels were significantly increased,the difference was statistically significant(all P<0.05);The pathological damage of kidney was aggravated.Compared with DMCLP group,the percentage of Ly6C hi monocytes in anti-PD-L1 group was reduced,the expression of MHCⅡon Ly6C hi monocytes significantly increased,the expression of inflammatory factors IL-6,TNF-αand IL-1

关 键 词：糖尿病 脓毒症 单核细胞 程序性死亡配体-1 盲肠结扎穿孔术 血清肌酐 

分 类 号：R459.7[医药卫生—急诊医学] R587.1[医药卫生—治疗学]