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
Effect of losartan on acute kidney injury and the relationship with mitochondrial fusion-fission in septic mice
王恩全 1阚宇飞 1孟姝岐 1宋宇 2谢克亮3
作者信息
- 1. 天津医科大学总医院麻醉科 天津市麻醉学研究所,天津 300052
- 2. 天津医科大学总医院重症医学科,天津 300052
- 3. 天津医科大学总医院麻醉科 天津市麻醉学研究所,天津 300052;天津医科大学总医院重症医学科,天津 300052
- 折叠
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.
Abstract
Objective To evaluate the effect of losartan on acute kidney injury(AKI)and the re-lationship 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 meth-od,and serum concentrations of tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6)and high-mob-ility 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 Drpl 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 tubular injury score were decreased,the ATP content and MMP were increased,the expression of Drpl was down-regulated,the expression of Mfn2 was up-regulated(P＜0.05),and the pathological changes of renal tissues were signifi-cantly attenuated in SA-AKI+LOS group.Conclusions Losartan can alleviate AKI in septic mice,and the mechanism may be related to promoting mitochondrial fusion and inhibiting mitochondrial fission.
关键词
氯沙坦/脓毒症/急性肾损伤/线粒体融合/线粒体分裂Key words
Losartan/Sepsis/Acute kidney injury/Mitochondrial fusion/Mitochondrial fis-sion引用本文复制引用
基金项目
国家自然科学基金(81971879)
天津市自然科学基金(17JCYBJC24800)
天津市科技计划重点研发计划科技支撑重点项目(18YFZCSY00560)
出版年
2024
NETL
NSTL
万方数据