核苷酸结合寡聚化结构域样受体蛋白3炎性小体在脓毒症急性肾损伤大鼠肾脏组织的表达及其影响  

Effect of nucleotide binding oligomerization domain-like receptor protein 3 inflammasome on kidney tissues in septic rats with acute kidney injury

作　　者：张建楠[1] 刘文[1] 昌广平[1] 曹延会[1] Zhang Jiannan;Liu Wen;Chang Guangping;Cao Yanhui(Department of Critical Care Medicine,the First Affiliated Hospital of Harbin Medical University,150001 Harbin,China)

机构地区：[1]哈尔滨医科大学附属第一医院重症医学科,哈尔滨150001

出　　处：《中华危重症医学杂志（电子版）》2020年第1期55-59,共5页Chinese Journal of Critical Care Medicine:Electronic Edition

基　　金：黑龙江省卫生计生委科研课题(2017-041)。

摘　　要：目的探讨核苷酸结合寡聚化结构域样受体蛋白(NLRP3)炎性小体在脓毒症急性肾损伤(AKI)大鼠中的作用及其机制。方法采用随机数字表法将18只雄性Wistar大鼠分为脂多糖(LPS)组、LPS+抑制剂组及阴性对照组3组,每组6只。LPS组大鼠给予腹腔内注射LPS(3.5 mg/kg),LPS+抑制剂组大鼠腹腔内注射LPS(3.5 mg/kg)及半胱氨酸天冬氨酸蛋白酶1(Caspase-1)抑制剂Belnacasan(VX-765,100 mg/kg),而阴性对照组大鼠则给予腹腔内注射0.3 mL等渗NaCl溶液。比较3组大鼠建模前及建模后24 h血清肌酐水平变化,采用酶联免疫吸附测定(ELISA)法检测3组大鼠建模24 h后血清白细胞介素1β(IL-1β)、IL-18及肿瘤坏死因子α(TNF-α)表达水平,采用Western-blotting检测3组大鼠建模24 h后NLRP3及Caspase-1蛋白表达水平。结果3组大鼠建模前后血清肌酐水平比较,差异有统计学意义(F=146.910,P<0.001)。进一步两两比较发现,建模后,LPS+抑制剂组及阴性对照组大鼠血清肌酐水平均较LPS组显著降低[(1.57±0.20)、(0.54±0.39)、(2.31±0.24)mg/L],且阴性对照组大鼠血清肌酐水平较LPS+抑制剂组更低(P均<0.05);而LPS组[(2.31±0.24)mg/L vs.(0.53±0.16)mg/L]及LPS+抑制剂组[(1.57±0.20)mg/L vs.(0.56±0.08)mg/L]大鼠建模后血清肌酐水平均较建模前显著升高(P均<0.05)。LPS组、LPS+抑制剂组及阴性对照组大鼠血清IL-1β[(9.33±1.16)、(1.93±0.30)、(1.88±0.30)ng/L]、IL-18[(23.8±2.8)、(19.6±1.5)(16.6±1.2)ng/L]、TNF-α[(42.3±2.1)、(23.9±2.5)、(23.5±1.3)ng/L]及NLRP3蛋白[(2.04±0.25)、(2.04±0.27)、(1.49±0.28)]和Caspase-1蛋白[(0.62±0.07)、(0.51±0.09)、(0.50±0.09)]表达水平比较,差异均有统计学意义(F=217.015、20.590、168.766、8.723、3.905,P均<0.05)。进一步两两比较发现,LPS+抑制剂组及阴性对照组大鼠血清IL-1β、IL-18、TNF-α表达水平均较LPS组大鼠显著降低,且阴性对照组大鼠IL-18表达水平较LPS+抑制剂组更低(P均<0.05)。阴性对照组大鼠Objective To investigate the role and mechanism of nucleotide binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome in septic rats with acute kidney injury(AKI).Methods A total of 18 male Wistar rats were randomly divided into a lipopolysaccharide(LPS)group,a LPS+inhibitor group,and a negative control group,6 in each group.Rats in the LPS group were given an intraperitoneal injection of LPS(3.5 mg/kg),and rats in the LPS+inhibitor group were given an intraperitoneal injection of LPS(3.5 mg/kg)and cysteinyl aspartate specific proteinase-1(Caspase-1)inhibitor Bernacasan(VX-765,100 mg/kg),while rats in the negative control group were given an intraperitoneal injection of isotonic NaCl solution(0.3 mL).Serum creatinine levels of rats in the 3 groups were compared before and after modeling for 24 h.The expression levels of serum interleukin-1beta(IL-1β),IL-18,and tumor necrosis factor-alpha(TNF-α)of rats in the 3 groups were measured by enzyme-linked immunosorbent assay(ELISA)after modeling for 24 h.In the meantime,the expression levels of NLRP3 and Caspase-1 proteins were detected by Western-blotting after modeling for 24 h.Results The serum creatinine levels of rats in the 3 groups were statistically significantly different before and after modeling(F=146.910,P<0.001).Further pairwise comparisons showed that after modeling,the serum creatinine levels in the LPS+inhibitor group and negative control group were both significantly lower than those in the LPS group[(1.57±0.20),(0.54±0.39),(2.31±0.24)mg/L],and the serum creatinine level in the negative control group[(0.54±0.39)mg/L vs.(1.57±0.20)mg/L]was lower than that in the LPS+inhibitor group,whereas the serum creatinine levels in the LPS group[(2.31±0.24)mg/L vs.(0.53±0.16)mg/L]and LPS+inhibitor group[(1.57±0.20)mg/L vs.(0.56±0.08)mg/L]after modeling were both significantly higher than those before modeling(all P<0.05).The expression levels of serum IL-1β[(9.33±1.16),(1.93±0.30),(1.88±0.30)ng/L],IL-18[(23.8±2.8),(19.6±1.5),(16.6±

关 键 词：脓毒症 核苷酸结合寡聚化结构域样受体蛋白3 急性肾损伤 

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