抑制组蛋白乳酸化修饰减轻高糖腹膜透析液诱导的腹膜炎症  

Inhibition of histone lactylation attenuated peritoneal in flamm ation induced by high-glucose peritoneal dialysate

作　　者：蔡青利 汪晓月 陈客宏 喻芳 Qingli Cai;Xiaoyue Wang;Kehong Chen;Fang Yu(Department of Nephrology,Daping Hospital,Army Military Medical University,Chongqing 400042,China;Chongqing Key Laboratory of Precision Diagnosis and Treatment for Kidney Diseases,Chongqing 400042,China)

机构地区：[1]陆军军医大学大坪医院肾脏内科,重庆400042 [2]重庆市肾脏疾病精准诊治重点实验室,400042

出　　处：《中华肾病研究电子杂志》2025年第1期34-43,共10页Chinese Journal of Kidney Disease Investigation(Electronic Edition)

基　　金：重庆市科技创新引导课题-院士专项(2022YSZX-JCX0007CSTB);重庆市自然科学基金重点项目(CSTB2023NSCQ-ZDX0008);国家自然科学基金青年项目(82200838)。

摘　　要：目的探讨组蛋白乳酸化修饰对高糖腹膜透析液诱导的腹膜炎症的作用。方法选取雄性8周龄C57BL/6小鼠32只,体重20~25 g,采用4.25%含糖腹透液和甲基乙二醛溶液(MGO)构建腹膜炎症小鼠模型,以乳酸化增强剂鱼藤酮(ROT)和乳酸化抑制剂草氨酸盐(OXA)作为干预因素。将小鼠随机分成对照组、高糖腹透液组、高糖腹透液+ROT组和高糖腹透液+OXA组:对照组不做任何处理;高糖腹透液组、高糖腹透液+ROT组和高糖腹透液+OXA组腹腔注射4.25%含糖腹透液(0.1 ml/g)与MGO(50μg/g),同时高糖腹透液+ROT组腹腔注射ROT(1 mg/kg),高糖腹透液+OXA组腹腔注射OXA(750 mg/kg),均1次/d。2周后结束实验,留取小鼠壁层腹膜组织,评估腹膜组织厚度,PAS染色观察腹膜组织病理变化,免疫组化染色评估炎症反应。细胞实验采用人腹膜间皮细胞系(HPMCs)细胞,分为对照组、高糖组(细胞培养基中加入高糖50%葡萄糖10.8μl/ml)、高糖+ROT组(培养基加入高糖和ROT 0.012 mg/m l)、高糖+OXA组(培养基加入高糖和OXA 0.45 mg/ml)。刺激48 h后收集细胞样本,检测各组细胞糖酵解关键酶的mRNA表达、糖酵解速率和三磷酸腺苷(ATP)浓度。小鼠腹膜组织和HPMCs细胞均采用免疫共沉淀法检测组蛋白乳酸化水平,酶联免疫吸附剂试验(ELISA)检测乳酸水平和炎症因子水平,实时荧光定量PCR检测炎症因子mRNA表达。结果动物实验结果显示,在小鼠腹膜厚度、乳酸水平和组蛋白H3乳酸化水平方面,以及在腹膜组织炎细胞浸润(巨噬细胞和中性粒细胞)和炎症因子(IL-1β和IL-6)表达水平方面,高糖腹透液组显著高于对照组、高糖腹透液+ROT组显著高于高糖腹透液组,而高糖腹透液+OXA组则低于高糖腹透液组(P均<0.05)。细胞实验显示,与对照组相比,高糖组的糖酵解关键酶、糖酵解速率、乳酸、组蛋白H3乳酸化和炎症因子(IL-1β和IL-6)水平显著增高,而ATP产生则显著降低(P均<0.05)。�Objective To investigate the role of histone lactylation in peritoneal inflammation induced by high-glucose peritoneal dialysate.Methods Thirty-two male C57BL/6 mice,aged 8 weeks and weighing 20-25 grams,were used to develop a peritoneal inflammation model by employing 4.25%glucose peritoneal dialysate and methylglyoxal(MGO)as inflammatory agents,but rotenone(ROT)and oxalate(OXA)as lactylation enhancer and inhibitor,respectively.The mice were divided into four groups:the control group,high-glucose dialysate group,high-glucose dialysate with ROT group(ROT group),and high-glucose dialysate with OXA(OXA group).The control group received no treatment,while the other three groups received daily intraperitoneal injection of 4.25%glucose dialysate(0.1 ml/g)and MGO(50μg/g).Additionally,the ROT group and OXA group also received 1 mg/kg ROT and 750 mg/kg OXA,respectively.The experimental period lasted for two weeks,after which the parietal peritoneal tissues were collected from the mice for evaluating the thickness of the peritoneal tissue,observing the pathological changes of the peritoneal tissue by PAS staining,and assessing the inflammatory response by immunohistochemical staining.For cell experiments,a cell line of human peritoneal mesothelial cells(HPMCs)was utilized.The cells were also divided into four groups:control culture group,high-glucose culture group,high-glucose culture plus ROT group,and high-glucose culture plus OXA group.Apart from the control culture group,the other three groups were treated with high-glucose of 50%glucose(10.8μl/ml),high-glucose plus ROT(0.012 mg/ml),and high-glucose plus OXA(0.45 mg/ml),respectively.After 48 hours of treatment,the cell groups were analyzed for mRNA levels of glycolytic enzymes,glycolytic rate,and ATP concentration.Both mice peritoneal tissues and HPMCs underwent immunoprecipitation for detecting histone lactylation,ELISA for detecting lactate and proteins of inflammatory factors,and real-time PCR for detecting mRNAs of inflammatory factors.Results Animal experiments

关 键 词：高糖腹膜透析液 腹膜间皮细胞 组蛋白乳酸化 腹膜炎症 

分 类 号：R73[医药卫生—肿瘤]