甘草酸对脓毒症急性肾损伤大鼠的保护作用及可能机制  

Protective effect of glycyrrhizic acid on sepsis induced acute kidney injury in rats and its possible mechanism

作　　者：周海银[1] 隆彩霞[1] 罗兰[1] 陈艳瑛 刘萍萍[1] 肖政辉[1] 张树菊 ZHOU Haiyin;LONG Caixia;LUO Lan;CHEN Yanying;LIU Pingping;XIAO Zhenghui;ZHANG Shuju(Emergency Center,Hunan Children’s Hospital,Hunan Province,Changsha,410007,China;Clinical Laboratory,Hunan Children’s Hospital,Hunan Province,Changsha,410007,China)

机构地区：[1]湖南省儿童医院急救中心,湖南长沙410007 [2]湖南省儿童医院检验科,湖南长沙410007

出　　处：《中国医药导报》2021年第28期24-28,39,共6页China Medical Herald

基　　金：湖南省卫生健康委科研立项课题(20200611);湖南省科技厅湖南省重点实验室、工程技术研究中心和科技基础条件平台组建立项项目(2018TP1028);湖南省重点研发计划项目(2018SK2135)。

摘　　要：目的研究甘草酸(GA)对脓毒症诱导大鼠急性肾损伤的保护作用及可能机制。方法SD大鼠分为假手术组、GA 50 mg/kg组、脓毒症组、脓毒症+GA 25 mg/kg组、脓毒症+GA 50 mg/kg组,每组6只。脓毒症组大鼠腹腔注射脂多糖(LPS)5 mg/kg建模。将HBZY-1细胞分为对照组、100μmol/L GA组、LPS组、LPS+GA 50μmol/L组和LPS+GA 100μmol/L组,LPS组用含10μg/ml LPS的培养基培养24 h构建肾炎模型。采用HE染色观察肾组织病理学变化;酶联免疫吸附试验法检测肾组织中肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)、细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)含量;Western blot检测各组HBZY-1细胞凋亡、炎症及氧化应激相关蛋白表达。结果脓毒症大鼠肾间质、肾小球和肾小管有炎症细胞浸润,部分肾小管细胞肿胀和空泡变性等改变,GA治疗后脓毒症大鼠肾组织上述病理改变明显减轻。脓毒症+GA 25 mg/kg组和和脓毒症+GA 50 mg/kg组大鼠肾组织中TNF-α、ICAM-1、VCAM-1表达水平低于脓毒症组(P<0.05);脓毒症+GA 50 mg/kg组大鼠肾组织MCP-1水平低于脓毒症组(P<0.05)。LPS+GA 50μmol/L组和LPS+GA 100μmol/L组HBZY-1细胞环氧化酶-2、核因子-κB蛋白表达水平低于LPS组(P<0.05或P<0.01);LPS+GA 100μmol/L组诱导型一氧化氮合酶水平低于LPS组(P<0.01)。LPS+GA 100μmol/L组HBZY-1细胞Bcl-2/Bax相对表达高于LPS组(P<0.01);LPS+GA 50μmol/L组和LPS+GA 100μmol/L组HBZY-1细胞活化半胱氨酸天冬氨酸蛋白酶-3蛋白表达低于LPS组(P<0.01)。LPS+GA 50μmol/L组和LPS+GA 100μmol/L组血红素加氧酶1蛋白表达水平高于LPS组(P<0.01);LPS+GA 100μmol/L组p-ERK蛋白表达水平高于LPS组(P<0.01)。结论GA对脓毒症诱导大鼠急性肾损伤具有保护作用,其机制可能与抑制细胞凋亡、氧化应激和炎症反应有关。Objective To study the protective effect and possible mechanism of glycyrrhizic acid(GA)on sepsis induced acute kidney injury in rats.Methods SD rats were divided into sham operation group,GA 50 mg/kg group,sepsis group,sepsis+GA 25 mg/kg group,and sepsis+GA 50 mg/kg group,with six rats in each group.Sepsis group rats were intraperitoneally injected with lipopolysaccharide(LPS)at 5 mg/kg.HBZY-1 cells were divided into control group,100μmol/L GA group,LPS group,LPS+GA 50μmol/L group and LPS+GA 100μmol/L group.LPS group was cultured with 10μg/ml LPS for 24 h to establish nephritis model.Histopathological changes of kidney were observed by HE staining;the contents of tumor necrosis factor-α(TNF-α),monocyte chemotactic protein-1(MCP-1),intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)in renal tissue were determined by enzyme-linked immunosorbent assay;HBZY-1 cells apoptosis,inflammation and oxidative stress related protein expression were analyzed by Western blot.Results The renal interstitium,glomerulus and tubules of sepsis rats were infiltrated with inflammatory cells,and some renal tubules were swollen and vacuolar degeneration.The above pathological changes in the renal tissues of sepsis rats were significantly relieved after GA treatment.The expression levels of TNF-α,ICAM-1 and VCAM-1 in rat renal tissues of sepsis+GA 25 mg/kg group and sepsis+GA 50 mg/kg group were lower than those of sepsis group(P<0.05).The level of MCP-1 in rat renal tissue of sepsis+GA 50 mg/kg group was lower than that of sepsis group(P<0.05).The expression levels of cyclooxygenase-2 and nuclear factor-κB protein in HBZY-1 cells in LPS+50μmol/L and LPS+100μmol/L GA groups were lower than those in LPS group(P<0.05 or P<0.01),the inducible nitric oxide synthase level in LPS+GA 100μmol/L group was lower than that in LPS group(P<0.01).The relative expression of Bcl-2/Bax in HBZY-1 cells in LPS+GA 100μmol/L group was higher than that in LPS group(P<0.01).The expression of cleaved Caspase-3 prot

关 键 词：甘草酸 大鼠 肾组织 凋亡 氧化应激 

分 类 号：R459.7[医药卫生—急诊医学]