瑞博西林对脓毒症急性肾损伤的肾脏保护作用及机制  

Protective effect and mechanism of Ribociclib on sepsis induced-acute kidney injury

作　　者：王倩[1] 龚晓莹[1] 贾佳[1] 李国福[1] Wang Qian;Gong Xiaoying;Jia Jia;Li Guofu(Department of Critical Care Medicine,Shengjing Hospital of China Medical University,Shenyang 110004,Liaoning,China)

机构地区：[1]中国医科大学附属盛京医院重症医学科,辽宁沈阳110004

出　　处：《中华危重病急救医学》2020年第2期204-209,共6页Chinese Critical Care Medicine

基　　金：国家自然科学基金(81471847);辽宁省自然科学基金(20180550267,2014021003);辽宁省沈阳市科技计划项目(F14-158-9-40)。

摘　　要：目的:探讨瑞博西林(Ribociclib)在脓毒症急性肾损伤(AKI)炎症反应中的作用及可能机制。方法:①按随机数字表法将20只成年雄性C57BL/6小鼠分为假手术组(Sham组;只开腹,不结扎盲肠、穿孔,假手术前12 h灌胃乳酸钠缓冲液)、瑞博西林对照组(只灌胃瑞博西林150 mg/kg)、盲肠结扎穿孔术(CLP)致脓毒症模型组(CLP组;CLP术前12 h灌胃乳酸钠缓冲液)和瑞博西林预处理组(CLP术前12 h灌胃瑞博西林150 mg/kg),每组5只。各组于术后12 h取肾组织,采用苏木素-伊红(HE)染色后观察肾组织病理学改变;采用酶联免疫吸附试验(ELISA)检测肾组织匀浆和细胞培养基上清液中炎性因子肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平;用蛋白质免疫印迹试验(Western Blot)检测周期相关蛋白磷酸化视网膜母细胞瘤蛋白(p-Rb)、凋亡相关蛋白Bcl-2和Bax表达。②用TCMK-1肾小管上皮细胞株进行体外验证实验,将细胞分为空白对照组、瑞博西林组(5μmol/L瑞博西林处理24 h)、脂多糖(LPS)组(200 mg/L的LPS培养基处理6 h)、瑞博西林+LPS组(用5μmol/L瑞博西林处理18 h后更换为含有5μmol/L瑞博西林和200 mg/L LPS的培养基联合处理6 h)。采用ELISA法检测上清液中炎性因子水平,用Western Blot检测细胞中p-Rb、Bcl-2、Bax、自噬相关蛋白微管关联蛋白1轻链3b(LC3bⅡ、LC3bⅠ)和p62、磷酸化蛋白激酶B(p-AKT)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)表达。结果:①动物实验显示,与Sham组相比,CLP可显著损伤小鼠肾脏组织,增加肾组织TNF-α、IL-6水平,降低肾组织p-Rb表达和Bcl-2/Bax比值;而瑞博西林对照组各指标与Sham组比较差异无统计学意义。与CLP组相比,瑞博西林预处理后小鼠肾组织损伤明显改善,病理学评分明显降低(分:1.48±0.16比2.68±0.16,P<0.01),肾组织炎性因子水平显著降低〔TNF-α(ng/g):340.55±34.96比745.08±58.86,IL-6(mg/g):17.33±1.01比114.20±20.49,均P<0.01〕,p-Rb表达Objective To investigate the role of Ribociclib in sepsis induced-acute kidney injury(AKI)and its possible mechanisms.Methods①Twenty adult male C57BL/6 mice were divided into sham operation group(Sham group;only open the abdomen without ligating or perforating the cecum,administered with sodium lactate buffer 12 hours before the sham operation),Ribociclib control group(administered with 150 mg/kg Ribociclib),cecal ligation and puncture(CLP)group(sepsis model induced by CLP;lactate buffer was given by intragastric administration 12 hours before CLP),and Ribociclib pretreatment group(administered with 150 mg/kg Ribociclib 12 hours before CLP)according to random number table,with 5 mice in each group.Kidneys were harvested 12 hours after the operation.Pathological changes in kidney were observed by hematoxylin-eosin(HE)staining.Tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)levels in mice kidney homogenate were measured by enzyme linked immunosorbent assay(ELISA).Western Blot was used to detect the expression of cell cycle-related protein phosphorylate retinoblastoma protein(p-Rb),apoptosis-related protein Bcl-2 and Bax.②Mouse renal tubular epithelial(TCMK-1)cell line was used for in vitro experiment.The cells were divided into control group,Ribociclib group(treated with 5μmol/L Ribociclib for 24 hours),lipopolysaccharide(LPS)group(treated with 200 mg/L LPS for 6 hours),Ribociclib+LPS group(replaced with the medium containing 5μmol/L Ribociclib and 200 mg/L LPS for 6 hours after exposing with 5μmol/L Ribociclib for 18 hours).Inflammatory cytokines in cell culture medium were detected by ELISA.The expression of p-Rb,Bcl-2 and Bax,autophagy-related proteins microtubule associated protein 1 light chain LC3b(LC3bⅡ,LC3bⅠ)and p62,phosphate protein kinase B(p-AKT),phosphorylated mammalian target of rapamycin(p-mTOR)were measured by Western Blot.Results①Animal experiments showed that,compared with the Sham group,the kidney tissue of mice were significantly damaged,the levels of TNF-αand IL-6 were incre

关 键 词：急性肾损伤 瑞博西林 脓毒症 

分 类 号：R459.7[医药卫生—急诊医学]