Caspase-1选择性抑制剂VX-765在大鼠急性肾损伤相关性肺损伤中的作用  

Effect of VX-765 as a selective inhibitor of caspase-1 on acute kidney injury associated lung injury in rats

作　　者：齐曼曼 王旭鹏[1] 郑维朝 白杨[1] 白晶 孙文波[1] QI Manman;WANG Xupeng;ZHENG Weichao;BAI Yang;BAI Jing;SUN Wenbo(Department of Anesthesiology,Cangzhou Central Hospital,Cangzhou 061000,China)

机构地区：[1]河北省沧州市中心医院麻醉科,061000

出　　处：《临床麻醉学杂志》2021年第6期629-634,共6页Journal of Clinical Anesthesiology

摘　　要：目的评价caspase-1选择性抑制剂VX-765在大鼠急性肾损伤(AKI)相关性肺损伤中的作用。方法SPF级SD雄性大鼠72只,9~10周龄,体重350~400 g。采用随机数字表法分为四组:假手术组(S组)、AKI组(A组)、DMSO+AKI组(D组)和VX-765+AKI组(V组),每组18只。S组暴露双肾45 min;A组采用切除右肾、夹闭左肾肾蒂45 min的方法建立AKI模型;D组建立AKI模型前经鼠尾静脉注射2%DMSO 0.5 ml/kg;V组建立AKI模型前经鼠尾静脉注射10%VX-7650.5 ml/kg。AKI模型建立24 h后取主动脉血样,采用酶偶联速率法与肌氨酸氧化酶法检测血清尿素氮(BUN)和肌酐(Scr)浓度,ELISA法检测血清IL-1β和IL-18浓度。随后处死大鼠取左肾组织和左肺组织。采用免疫荧光法检测肾组织caspase-1/DAPI双阳性细胞百分比,Western blot法检测肾组织cleaved GSDMD蛋白及肺组织ZO-1蛋白含量,Paller评分法评估HE染色后的肾小管损伤情况,烘干法检测肺部干湿比重,计算肺含水率,ELISA法检测肺伊文斯蓝评估肺血管通透性,肺损伤评分评估MASSON染色后的肺组织损伤程度。结果与S组比较,A组、D组、V组血清BUN、Scr、IL-1β和IL-18浓度明显升高,肾组织caspase-1/DAPI双阳性细胞百分比明显升高,肾组织caspase-1和cleaved GSDMD蛋白含量明显升高,肾损伤Paller评分明显升高,肺组织ZO-1蛋白含量明显降低,肺含水率、肺血管通透性、肺损伤评分明显升高(P<0.05)。与A组比较,V组血清BUN、Scr、IL-1β和IL-18浓度明显降低,肾组织caspase-1/DAPI双阳性细胞百分比明显降低,肾组织caspase-1和cleaved GSDMD蛋白含量明显降低,肾损伤Paller评分明显降低,肺组织ZO-1蛋白含量明显升高,肺含水率、肺血管通透性、肺损伤评分明显降低(P<0.05)。A组、D组各指标差异无统计学意义。结论Caspase-1选择性抑制剂VX-765可以降低血清中BUN、Scr、IL-1β和IL-18浓度,减少肾组织细胞焦亡,改善肺水肿及肺血管通透性,减轻AKI相关性肺损伤�Objective To evaluate the effect of VX-765,a selective inhibitor of caspase-1,on acu￣te kidney injury(AKI)associated lung injury in rats.Methods Seventy-two adult male SD rats,aged 9-10 weeks,weighing 350-400 g,were randomly divided into four groups:sham group(group S),AKI group(group A),DMSO+AKI group(group D)and VX-765+AKI group(group V).The AKI model was established by excising the right kidney and clamping the pedicle of the left kidney for 45 minutes.VX-765(0.5 ml/kg)and 2%DMSO(0.5 ml/kg)were injected through tail vein respectively in group D and group V 1 hour before AKI.The kidneys were exposed for 45 minutes under anesthesia in sham-operated animals.Twenty-four hours post-AKI,renal pathological changes,pulmonary vascular permeability,lung wet/dry weight were measured.Blood urea nitrogen(BUN)and serum creatinine(Scr)were detected with enzyme coupling rate method and sarcosine oxidase method.IL-1βand IL-18 in blood serum were evaluated by enzyme linked immunosorbent assay(ELISA).The expressions of caspase-1 in kidney 24 hours after AKI were assessed by immunofluorescence assay,the expressions of cleaved GSDMD in the kidney and ZO-1 in the lung were detected by western blot.Results Compared with group S,serum concentrations of BUN,Scr,IL-1βand IL-18 were significantly increased,the expression of caspase-1 and cleaved GSDMD in renal tissue,renal cortex injury scores,Evans blue content in lung tissue and lung injury scores were also significantly increased in groups A,D,and V.But the expression of ZO-1 were statistically decreased(P<0.05).Compared with group A,serum BUN,Scr,IL-1β,IL-18,caspase-1 and cleaved GSDMD expression,renal cortex injury scores,Evans blue content in lung tissue and lung injury scores were all significantly decreased,and the expression of protein ZO-1 were statistically increased in group V(P<0.05).There was no significant difference between group A and group D.Conclusion VX-765,a selective inhibitor of caspase-1,can alleviate renal cell pyroptosis and AKI-related lung injury by redu

关 键 词：急性肾损伤 肺损伤 细胞焦亡 CASPASE-1 

分 类 号：R692[医药卫生—泌尿科学] R563[医药卫生—外科学]