廖豪峰?庞梦雅?张政?葛缅

【摘要】 目的 探討p53上调凋亡调控因子（PUMA）是否参与调节高脂饮食（HFD）所致糖尿病小鼠脂多糖（LPS）诱导的急性肾损伤。方法 将24只小鼠随机均分为普食（Chow）组与HFD组，分别以普通饲料或者高脂饮食喂养12周，饮食干预结束后分别再分为LPS组与Con组，LPS组小鼠腹腔注射LPS 10 mg/kg制备脓毒症急性肾损伤模型，Con组给予等体积磷酸盐缓冲液腹腔注射，24 h后检测小鼠肾病理损伤、血清肌酐、血尿素氮、PUMA蛋白和Caspase-3蛋白表达水平以及肾脏凋亡情况。结果 与Chow组相比，HFD组小鼠体质量增加、随机血糖升高（P均< 0.05），产生胰岛素抵抗。与Con组相比，LPS组血清肌酐和血尿素氮升高（P均< 0.05），出现肾上皮细胞形态异常、上皮细胞坏死和炎症浸润等病理损伤，原位末端转移酶标记技术染色显示肾组织出现大量凋亡细胞，PUMA蛋白水平及Caspase-3蛋白表达增高（P均< 0.05）。与Chow+LPS组相比，HFD+LPS组上述表现更为明显（P均< 0.05）。结论 HFD通过上调PUMA表达，促进了LPS诱导的小鼠急性肾损伤以及凋亡。

【关键词】 p53上调凋亡调控因子;高脂饮食;糖尿病;脂多糖;急性肾损伤

Effect of PUMA on LPS-induced acute kidney injury and apoptosis in high-fat diet-fed mice Liao Haofeng， Pang Mengya， Zhang Zheng， Ge Mian. Department of Anesthesiology， the Third Affiliated Hospital of Sun Yat-sen University， Guangzhou 510630， China

Corresponding author， Ge Mian， E-mail： gemiansums@ 163. com

【Abstract】 Objective To investigate whether p53 upregulated modulator of apoptosis （PUMA） participates in regulating the acute kidney injury induced by lipopolysaccharide （LPS） in high-fat diet （HFD）-fed mice. Methods Twenty-four mice were randomly divided into the Chow and HFD groups， and fed with regular chow or HFD for 12 weeks， respectively. Subsequently， the animals were further divided into the control and LPS groups. Mice in the LPS group were injected with LPS 10 mg/kg intraperitoneally to establish the mouse models of acute kidney injury， whereas an equivalent amount of PBS was administered in the control group. The expression levels of serum creatinine， urea nitrogen， PUMA protein and Caspase-3 protein were detected and the pathological kidney injury and apoptosis were observed 24 h later. Results Compared with the Chow group， mice in the HFD group had significantly higher body weight （P < 0.05）， elevated random blood glucose （P < 0.05） and insulin resistance. Compared with the control group， the serum levels of creatinine and urea nitrogen of mice in the LPS group were significantly increased （both P < 0.05）， and presented with obvious pathological damages， such as morphological abnormality of renal epithelial cells， epithelial cell necrosis and inflammatory infiltration. Meanwhile， in situ terminal deoxynucleotidyl transferase revealed a large quantity of apoptotic cells and higher PUMA protein and Caspase-3 protein levels were observed in the renal tissues of mice in the LPS group （both P < 0.05）. Compared with the Chow+LPS group， the findings above in the HFD + LPS group were more pronounced （all P < 0.05）. Conclusion HFD can aggravate LPS-induced acute kidney injury and apoptosis probably by up-regulating the expression level of PUMA.

【Key words】 p53 upregulated modulator of apoptosis（PUMA）;High-fat diet;Diabetes mellitus;

Lipopolysaccharide;Acute kidney injury

近年来，由于超重和肥胖率的上升以及体育活动的减少，我国的2型糖尿病发病率迅速上升，已经成为公共卫生的主要挑战[1-2]。多项研究表明，肥胖以及2型糖尿病患者在临床及围术期中发生感染性并发症的风险更高，这可能导致脓毒症发生率增高以及ICU脓毒症相关患者的病死率增加[3-5]。脓毒症是宿主对感染的反应失调导致的危及生命的器官功能障碍[6]。肾脏是脓毒症发展过程中最容易受损的器官[4-5， 7]。脓毒症相关急性肾损伤（S-AKI）在ICU较为常见，同时AKI会导致脓毒症患者病死率增高。与ICU其他损伤相比，S-AKI病死率更高[8-9]。此外，有研究表明糖尿病是术后感染、S-AKI的独立危险因素[10-11]。因此，关注糖尿病患者的S-AKI成为急需解决的临床问题，然而目前研究并不多。

既往研究显示，多种因素参与了S-AKI的发病机制，比如炎症损伤、氧化应激、凋亡等，其中凋亡被证明起了重要作用[12-13]。……