乙酸盐通过G蛋白耦联受体43减轻脓毒症大鼠急性肾损伤的作用及机制研究  

Effect and related mechanism of acetate in alleviating acute kidney injury in septic rats through G-protein coupled receptor 43

作　　者：史星宇 邢家瑜 王毅[1] 李建[1] 柴瑞峰[1] 于湘友[1] Shi Xingyu;Xing Jiayu;Wang Yi;Li Jian;Chai Ruifeng;Yu Xiangyou(Department of Critical Medicine Center,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学第一附属医院重症医学科,乌鲁木齐830054

出　　处：《中华危重病急救医学》2024年第11期1147-1152,共6页Chinese Critical Care Medicine

基　　金：新疆维吾尔自治区科技支疆项目(2021E02064);新疆维吾尔自治区自然科学基金资助项目(2021D01C306,2020D01C233)。

摘　　要：目的探讨乙酸盐对脓毒症急性肾损伤(AKI)大鼠的保护作用及机制。方法按照随机数字表法将雄性SD大鼠分为假手术组(Sham组)、盲肠结扎穿孔术致脓毒症组(CLP组)、乙酸盐预处理组〔NaA组,CLP术前灌胃乙酸钠(NaA)300 mg/kg,每日2次,连续7 d〕,每组7只。于制模后24 h腹主动脉取血并处死大鼠取肾脏组织,用酶联免疫吸附试验(ELISA)检测血清中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、肾损伤分子-1(KIM-1)水平;液相色谱法检测血清乙酸盐浓度;用硫代巴比妥酸法检测肾组织丙二醛(MDA)水平;用比色法测定肾组织髓过氧化物酶(MPO)水平;苏木素-伊红(HE)染色后光镜下观察肾组织病理学改变并进行肾小管损伤评分;蛋白质免疫印迹试验(Western blotting)检测肾组织中G蛋白耦联受体43(GPR43)蛋白以及腺苷酸活化蛋白激酶/沉默信息调节因子1/过氧化物酶体增殖物激活受体γ辅激活因子1α(AMPK/SIRT1/PGC-1α)通路相关蛋白的表达;免疫组化法检测肾组织中GPR43、磷酸化AMPK(p-AMPK)、SIRT1、PGC-1α的阳性表达。结果与Sham组相比,CLP组血清IL-6、TNF-α、KIM-1水平明显升高,肾组织中MDA、MPO含量明显升高,血清乙酸盐水平明显下降;HE染色显示大部分肾小管上皮细胞变性,局部伴坏死,大量刷状缘损伤、脱落,肾小管结构破坏、碎裂,肾间质较多炎症细胞浸润,肾小管损伤评分明显升高;肾脏组织中GPR43、p-AMPK、SIRT1、PGC-1α表达均明显降低,提示脓毒症大鼠出现肾损伤,且氧化应激和炎症水平升高。与CLP组相比,NaA组血清IL-6、TNF-α、KIM-1水平明显降低〔IL-6(ng/L):126.20±6.23比161.00±17.37,TNF-α(ng/L):85.59±7.70比123.50±17.78,KIM-1(μg/L):2.92±0.38比4.73±0.36,均P<0.05〕,肾组织中MDA、MPO含量明显降低〔MDA(μmol/g):6.56±0.18比8.53±0.34,MPO(U/g):2.99±0.20比3.72±0.29,均P<0.05〕,HE染色结果显示肾脏损伤有所缓解,肾小管损伤评分明显下降〔分:1(1,2Objective To explore the protective effect and mechanism of acetate on sepsis-induced acute kidney injury(AKI)in rats.Methods Male Sprague-Dawley(SD)rats were divided into sham operation group(Sham group),sepsis group caused by cecal ligation and puncture(CLP group),and acetate pretreatment group[NaA group,gavage sodium acetate(NaA)300 mg/kg twice a day for 7 consecutive days before CLP]using a random number table method,with 7 rats in each group.The blood was taken from the main abdominal artery 24 hours after modeling,and renal tissue was collected from the rats.Enzyme linked immunosorbent assay(ELISA)was used to detect the serum levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and kidney injury molecule-1(KIM-1).The concentration of serum acetate was determined by high performance liquid chromatography.The level of malondialdehyde(MDA)in renal tissue was detected by thiobarbituric acid method.Myeloperoxidase(MPO)in renal tissue was detected by colorimetric method.Hematoxylin-eosin(HE)staining was used to observe histopathological changes and assess renal tubule injury score.Western blotting was used to detect the protein expressions of G-protein coupled receptor 43(GPR43)and adenosine monophosphate-activated protein kinase/silence infor-mation regulator 1/peroxlsome proliferator-activated receptor-γcoactlvator-1α(AMPK/SIRT1/PGC-1α)pathway.The positive expressions of GPR43,phosphorylation-AMPK(p-AMPK),SIRT1,PGC-1αwere detected by immunohistochemistry.Results Compared with Sham group,the serum levels of IL-6,TNF-αand KIM-1 were significantly increased in CLP group,the contents of MDA and MPO in renal tissue were increased,and the content of acetate was significantly decreased.HE staining results showed that most of the tubular epithelial cells were denaturated with local necrosis,a large number of brush border injuries and shedding,tubular structure destruction and fragmentation,and more inflammatory cells infiltrated the renal interstitium,the renal tubular injury score significantly increase

关 键 词：脓毒症 急性肾损伤 短链脂肪酸 氧化应激 G蛋白耦联受体43 

分 类 号：R459.7[医药卫生—急诊医学] R692[医药卫生—治疗学]