摘要
目的探讨七氟烷对大鼠肾缺血再灌注损伤的影响及其作用机制。方法选取6~8周龄、体质量(250±30)g雄性SD大鼠40只,按数字表法随机分成假手术组(Sham组)、肾缺血再灌注组(IR组)、七氟烷预处理组(Sev组)、七氟烷预处理+沉默信号调节因子1(SIRT1)抑制剂组(EX组)4组,每组10只。肾缺血再灌注损伤模型制备前2天和术前10 min,Sham组、IR组、Sev组大鼠腹腔注射1%二甲基亚砜(DMSO)溶液(5 mL/kg),EX组腹腔注射含抑制剂EX 527(1 mg/mL)的1%DMSO溶液(5 mL/kg)。Sham组:切除右侧肾脏,暴露左侧肾脏但不夹闭肾蒂;IR组:切除右侧肾脏,制备左侧肾脏缺血再灌注模型;Sev组、EX组:先吸入七氟烷45 min后,再按IR组方法制备缺血再灌注模型。于制备缺血再灌注模型术后3 h,收集各组大鼠肾脏组织与左心室血液,取材后以颈椎脱位法处死大鼠。观察项目:(1)检测各组大鼠左心室血液血清肌酐(Scr)、尿素氮(BUN)。(2)制备各组大鼠肾脏组织病理切片,光镜下观察肾组织病理变化,对肾小管按Paller评分标准进行评分,评估肾脏损伤情况。(3)采用原位末端脱氧核苷酸转移酶介导的dUTP切口末端标记技术(TUNEL)检测各组大鼠肾脏组织细胞凋亡情况。(4)取各组大鼠肾脏组织病理切片,在透射电子显微镜下观察肾脏组织自噬情况。(5)采用蛋白质印迹法(Western blotting)检测各组大鼠肾脏组织SIRT1、叉头盒转录因子O3(FoxO3)蛋白,以及凋亡蛋白(BAX/Bcl 2)、自噬相关蛋白(Beclin1、LC3A/B)的表达水平。结果(1)Sham组大鼠血清Scr和BUN分别为(50.74±5.91)μmol/L和(10.11±0.80)mmol/L,IR组分别为(90.18±11.22)μmol/L和(53.39±6.29)mmol/L,Sev组分别为(63.70±8.69)μmol/L和(27.68±3.41)mmol/L,EX组分别为(80.18±9.15)μmol/L和(33.20±3.57)mmol/L。与Sham组相比,IR组、Sev组、EX组血清Scr、BUN水平均升高;与IR组相比,Sev组、EX组血清Scr、BUN水平均下降;与Sev组相比,EX组血清Scr、BUN水平均升高:
Objective This study aimed to investigate the effect of sevoflurane on renal ischemia‑reperfusion injury in rats and its mechanism.Methods Forty male SD rats,aged 6‑8 weeks old with a body weight of(250±30)g,were randomly divided into four groups:the sham surgery group(Sham group),renal ischemia‑reperfusion group(IR group),sevoflurane pretreatment group(Sev group),and sevoflurane pretreatment+silencing signal regulator 1(SIRT1)inhibitor group(EX group),with 10 rats in each group.Two days before the preparation of the renal ischemia‑reperfusion injury model and 10 min before surgery,the rats in the Sham,IR,and Sev groups were intraperitoneally injected with 1%dimethyl sulfoxide(DMSO)solution(5 mL/kg),and the EX group was intraperitoneally injected with 1%DMSO solution(5 mL/kg)containing the EX-527 inhibitor(1 mg/mL).In the Sham group,the right kidney was removed,and the left kidney was exposed but not clamped.In the IR group,the right kidney was resected,and an left renal ischemia‑reperfusion model was established.In the Sev group and EX group,sevoflurane was inhaled for 45 min,and then an ischemia‑perfusion model was prepared in accordance with the method used in the IR group.Three hours after the preparation of the ischemia‑reperfusion model,renal tissue and left ventricular blood of each group were collected,and the rats were sacrificed by cervical dislocation after the material was collected.Observation was performed as follows:(1)Serum creatinine(Scr)and urea nitrogen(BUN)were detected in the left ventricle of rats in each group.(2)Pathological sections of kidney histopathology of each group of rats were prepared;pathological changes in kidney tissue were observed under light microscopy,and renal tubules were scored in accordance with the Paller scoring standard to evaluate kidney damage.(3)In-situ terminal deoxynucleotidyl transferase-mediated dUTP incision-end labeling technology was used to detect apoptosis in kidney tissue cells of rats.(4)The pathological sections of kidney tissue in each
作者
杨传铭
易铭
韩冰
张一粟
李晓红
Yang Chuanming;Yi Ming;Han Bing;Zhang Yisu;Li Xiaohong(Department of Anaesthesiology,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China;Graduate School of Bengbu Medical College,Bengbu 233030,China)
出处
《中华解剖与临床杂志》
2023年第11期749-757,共9页
Chinese Journal of Anatomy and Clinics
基金
安徽省高校省级自然科学研究重点项目(KJ2017A246,KJ2020A0581)
蚌埠医学院研究生科研创新计划项目(Byycxz20019)。
关键词
再灌注损伤
七氟烷
肾损伤
大鼠
模型
动物
自噬
沉默信号调节因子1
叉头盒转录因子O3
Reperfusion injury
Sevoflurane
Kidney injury
Rats
Models,animal
Autophagy
Silent information regulator sirtuin 1
Forkhead transcription factor O3
