N-乙酰半胱氨酸对脓毒症小鼠急性肾损伤的保护作用及机制研究  

Protective effect and mechanism of N-acetylcysteine on acute kidney injury in septic mice

作　　者：樊恒[1] 乐健伟[1] 叶继辉[1] 孙敏[1] 朱建华[1] Fan Heng;Le Jianwei;Ye Jihui;Sun Min;Zhu Jianhua(Department of Intensive Care Unit,Ningbo First Hospital,Ningbo 315010,China)

机构地区：[1]宁波市第一医院重症医学科,浙江宁波315010

出　　处：《中华危重症医学杂志（电子版）》2021年第3期180-186,共7页Chinese Journal of Critical Care Medicine:Electronic Edition

基　　金：浙江省自然科学基金项目(LQ18H150001);浙江省医药卫生科技计划项目(2020KY815、2020KY828)。

摘　　要：目的探讨N-乙酰半胱氨酸(NAC)对脓毒症小鼠急性肾损伤(AKI)的保护作用及其机制。方法将60只小鼠分为假手术组、脓毒症AKI组(SAKI组)和SAKI+NAC组,每组各20只。采用盲肠结扎穿刺法(CLP)构建SAKI小鼠模型,其中SAKI+NAC组在使用NAC(50 mg/kg)预处理3 d后行CLP。假手术组小鼠仅进行外科手术,不予CLP。模型构建成功后24 h处死小鼠,采用苏木素-伊红(HE)染色法评估小鼠肾组织病理损伤,比较3组小鼠血清肌酐、血尿素氮、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-6、病理学评分、丙二醛、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶、肾组织细胞凋亡情况以及Toll样受体4(TLR4)、核因子κB(NF-κB)、Caspase-3、Bax蛋白表达水平。结果 HE染色结果显示,假手术组小鼠肾组织切片中肾小球、肾小管和血管结构清晰正常;SAKI组小鼠可见肾小球破裂、广泛的肾小管坏死和刷状缘的丧失,间质大量炎症细胞浸润;SAKI+NAC组小鼠肾组织损伤明显减轻,肾小球、肾小管上皮细胞可见充血水肿,间质炎症细胞明显减少。假手术组、SAKI组和SAKI+NAC组小鼠血清肌酐[(39.0±1.8)、(129.6±4.8)、(81.7±2.0)μmol/L]、血尿素氮[(6.65±0.29)、(24.78±0.71)、(15.45±0.40)mmol/L]、TNF-α[(16.8±0.4)、(43.5±1.0)、(22.0±0.6)ng/L]、IL-1β[(15.2±0.7)、(44.8±1.0)、(25.5±0.8)ng/L]、IL-6 [(20.8±0.4)、(70.6±2.0)、(35.8±1.2)ng/L]、病理学评分[(0.65±0.11)、(3.55±0.14)、(2.00±0.15)分]、丙二醛[(25.1±0.7)、(85.3±1.1)、(40.1±1.6)nmol/g]、SOD [(90.4±1.4)、(22.4±0.8)、(70.7±2.5)U/mg]、GSH-Px [(336.2±5.5)、(181.1±2.5)、(265.0±6.4)U/mg]、过氧化氢酶[(14.9±0.6)、(5.7±0.4)、(8.4±0.4)U/mg]、TUNEL阳性细胞数[(1.35±0.20)、(13.55±0.72)、(8.25±0.42)个]以及TLR4 [(0.14±0.03)、(0.40±0.03)、(0.18±0.03)]、NF-κB [(0.20±0.04)、(0.66±0.05)、(0.16±0.03)]、Caspase-3 [(0.152±0.027)、(0.464±0.061)、(0.147±0.022)]Objective To investigate the protective effect and mechanism of N-acetylcysteine(NAC)on acute kidney injury(AKI)in septic mice.Methods Sixty mice were divided into a sham operation group(sham group),a septic AKI group(SAKI group)and a SAKI+NAC group,with 20 mice in each group.The SAKI mouse model was constructed by cecal ligation and puncture(CLP),and mice in the SAKI+NAC group were performed CLP after pretreatment with NAC(50 mg/kg)for 3 d.Mice in the sham group only underwent surgery without CLP.All mice were sacrificed when the model was successfully constructed for 24 h,and the renal tissue pathological injury was evaluated by hematoxylin-eosin(HE)staining.The serum creatinine,blood urea nitrogen,tumor necrosis factor-alpha(TNF-α),interleukin-1 beta(IL-1β),IL-6,pathological score,malondialdehyde,superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase and renal tissue apoptosis were compared among these three groups,as well as protein levels of Toll-like receptor 4(TLR4),nuclear factor-kappa B(NF-κB),Caspase-3 and Bax.Results HE staining showed that the glomerular,tubular and vascular structures were clear and normal in the renal tissue sections of mice in the sham group.The glomerular rupture,extensive tubular necrosis,loss of brush border and a large number of interstitial inflammatory cells were seen in the SAKI group.The renal tissue damage was alleviated,the glomerular and renal tubular epithelial cells revealed hyperemia and edema,and the interstitial inflammatory cells significantly decreased in the SAKI+NAC group.The serum creatinine[(39.0±1.8),(129.6±4.8),(81.7±2.0)μmol/L],blood urea nitrogen[(6.65±0.29),(24.78±0.71),(15.45±0.40)mmol/L],TNF-α[(16.8±0.4),(43.5±1.0),(22.0±0.6)ng/L],IL-1β[(15.2±0.7),(44.8±1.0),(25.5±0.8)ng/L],IL-6[(20.8±0.4),(70.6±2.0),(35.8±1.2)ng/L],pathological score[(0.65±0.11),(3.55±0.14),(2.00±0.15)],malondialdehyde[(25.1±0.7),(85.3±1.1),(40.1±1.6)nmol/g],SOD[(90.4±1.4),(22.4±0.8),(70.7±2.5)U/mg],GSH-Px[(336.2±5.5),(181.1±2.5),(265.0±6.4)U

关 键 词：N-乙酰半胱氨酸 脓毒症 急性肾损伤 小鼠 

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