摘要
目的评价氯沙坦对脓毒症小鼠急性肾损伤的影响及其与线粒体融合-分裂的关系。方法 SPF级雄性C57BL/6J小鼠128只,6~8周龄,体质量20~25 g,采用随机数字表法分为4组(n=32):假手术组(Sham组)、假手术+氯沙坦组(Sham+LOS组)、脓毒症相关性急性肾损伤组(SA-AKI组)及脓毒症相关性急性肾损伤+氯沙坦组(SA-AKI+LOS组)。采用盲肠结扎穿孔法建立小鼠脓毒症模型。Sham+LOS组及SA-AKI+LOS组分别于假手术或造模前3 d开始,腹腔注射氯沙坦5 mg/kg,1次/d,连续3 d;Sham组及SA-AKI组腹腔注射等量溶剂。随机取20只小鼠,观察术后7 d生存情况。于假手术或造模后24 h,采用比色法测定血清BUN及Cr浓度,采用ELISA法测定血清TNF-α、IL-6和高迁移率族蛋白B1(HMGB1)浓度;取肾组织,HE染色后光镜下观察病理学结果,并行肾小管损伤评分,采用荧光素酶法测定ATP含量,采用JC-1法测定线粒体膜电位(MMP)水平,采用Western blot法检测动力相关蛋白1(Drp1)及线粒体融合蛋白2(Mfn2)表达。结果与Sham组比较,SA-AKI组及SA-AKI+LOS组生存率降低,血清BUN、Cr、TNF-α、IL-6、HMGB1浓度和肾小管损伤评分升高,肾组织ATP含量及MMP下降,Drp1表达上调,Mfn2表达下调(P<0.05),肾组织发生病理学损伤;与SA-AKI组比较,SA-AKI+LOS组生存率升高,血清BUN、Cr、TNF-α、IL-6、HMGB1浓度和肾小管损伤评分下降,肾组织ATP含量及MMP升高,Drp1表达下调,Mfn2表达上调(P<0.05),肾组织病理学损伤减轻。结论氯沙坦可减轻脓毒症小鼠急性肾损伤,机制可能与促进线粒体融合,抑制线粒体分裂有关。
Objective To evaluate the effect of losartan on acute kidney injury(AKI)and the relationship with mitochondrial fusion-fission in septic mice.Methods One hundred and twenty-eight SPF male C57BL/6J mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups(n=32 each)using a random number table method:sham operation group(Sham group),sham operation+losartan group(Sham+LOS group),sepsis-associated AKI group(SA-AKI group),and sepsis-associated AKI+losartan group(SA-AKI+LOS group).Sepsis was induced by cecal ligation and puncture in anesthetized mice.Sham+LOS group and SA-AKI+LOS group received intraperitoneal injection of losartan 5 mg/kg,once a day,for 3 consecutive days,starting from 3 days before sham operation or developing the model.The equal volume of solvent was given instead in Sham group and SA-AKI group.Twenty mice were randomly selected to observe the survival 7 days after surgery.At 24 h after sham operation or establishing the model,serum blood urea nitrogen(BUN)and creatinine(Cr)concentrations were determined by the colorimetric method,and serum concentrations of tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6)and high-mobility group box 1 protein(HMGB1)were measured using enzyme-linked immunosorbent assay.Renal tissues were obtained for microscopic examination of pathological changes which were scored and for determination of mitochondrial membrane potential(using JC-1 method)and expression of dynamin-related protein 1(Drp1)and mitofusin-2(Mfn2)(using Western blot).Results Compared with Sham group,the survival rate was significantly decreased,the serum BUN,Cr,TNF-α,IL-6 and HMGB1 concentrations and renal tubular injury score were increased,the ATP content and MMP were decreased,the expression of Drp1 was up-regulated,the expression of Mfn2 was down-regulated(P<0.05),and pathological changes were found in renal tissues in SA-AKI group and SA-AKI+LOS group.Compared with SA-AKI group,the survival rate was significantly increased,serum concentrations of BUN,Cr,TNF-α,IL-6 and HMGB1 and renal t
作者
王恩全
阚宇飞
孟姝岐
宋宇
谢克亮
Wang Enquan;Kan Yufei;Meng Shuqi;Song Yu;Xie Keliang(Department of Anesthesiology,Tianjin Medical University General Hospital,Tianjin Research Institute of Anesthesiology,Tianjin 300052,China;Department of Critical Care Medicine,Tianjin Medical University General Hospital,Tianjin 300052,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2024年第2期220-224,共5页
Chinese Journal of Anesthesiology
基金
国家自然科学基金(81971879)
天津市自然科学基金(17JCYBJC24800)
天津市科技计划项目重点研发计划科技支撑重点项目(18YFZCSY00560)。
关键词
氯沙坦
脓毒症
急性肾损伤
线粒体融合
线粒体分裂
Losartan
Sepsis
Acute kidney injury
Mitochondrial fusion
Mitochondrial fission
