摘要
目的 探讨右美托咪定对异丙酚诱导的发育期大鼠海马组织细胞焦亡的作用及机制.方法 40只健康新生7日龄Sprague-dawley(SD)大鼠随机均分为对照(Con)组(腹腔注射等容量的生理盐水)、异丙酚(Pro)组(腹腔注射异丙酚50 mg/kg)、右美托咪定预先给药(DP)组(腹腔注射右美托咪定25 μg/kg+Pro组方案)、A激酶锚定蛋白150(AKAP150)腺病毒+DP(ADP)组(AKAP150腺病毒腹腔注射+DP组方案),Con组大鼠于注射结束后2 h、其余组大鼠于麻醉苏醒后2 h分别处死并取海马组织,采用透射电镜下观察海马组织超微结构特点,采用蛋白印迹实验(Western blot)和实时荧光定量PCR(RT-qPCR)检测海马组织AKAP150、蛋白激酶A(PKA)、NOD 样受体家族 Pyrin 域蛋白-3(NLRP3)、Gasdermin-D 蛋白(GSDMD)、白细胞介素-1β(IL-1β)及白细胞介素-18(IL-18)蛋白和mRNA的表达.结果 Pro组、ADP组大鼠海马组织细胞膜破损形成孔道,DP组细胞膜结构基本完整;与Con组比较,Pro组、DP组及ADP组大鼠海马组织AKAP150、PKA表达下调,NLRP3、GSDMD、IL-1β、IL-18表达上调,差异均有统计学意义(P＜0.05);与Pro组比较,DP组大鼠海马组织AKAP150、PKA表达上调,NLRP3、GSDMD、IL-1β及IL-18表达下调,差异均有统计学意义(P＜0.05);与DP组比较,ADP组大鼠海马组织AKAP150、PKA表达下调,NLRP3、GSDMD、IL-1β及IL-18表达上调,差异均有统计学意义(P＜0.05).结论 右美托咪定可减轻异丙酚诱导的发育期大鼠海马组织的细胞焦亡,其机制可能与激活AKAP150表达、增强PKA活化、抑制细胞焦亡相关蛋白表达以及炎性因子IL-1β、IL-18释放有关.
Abstract
Objective To explore the effect dexmedetomidine on propofol-induced pyroptosis in rat hippocampal tissue and its mode of action at developmental stage.Methods Forty healthy 7-day old Sprague-Dawley(SD)rats were randomly assigned to control(Con)group(intraperitoneal injection of equal volume of normal saline),propofol(Pro)group(intraperitoneal injection of propofol 50 mg/kg),dexmedetomidine pretreatment(DP)group(intraperitoneal injection of dexmedetomidine 25 μg/kg+dosing regimen for Pro group),ADP group[intraperitoneal injection of adenovirus carrying A-kinase anchoring protein 150(AKAP150)+DP group regimen].The rats were sacrificed and hippocampal tissues were taken at 2 h after injection for Con group or at 2 h after awakening from anesthesia for the remaining groups.Transmission electron microscopy was used to observe ultrastructural characteristics of hippocampal tissues.Western blot and real-time fluorescence quantitative PCR(RT-qPCR)were used to detect the protein and mRNA expressions of AKAP150,protein kinase A(PKA),NOD-like receptor family Pyrin domain protein-3(NLRP3),gasdermin-D protein(GSDMD),interleukin-1β(IL-1β),and IL-18 in hippocampal tissues.Results Cell membranes of rat hippocampal tissues in Pro and ADP groups were damaged and pored,while cell membrane structure in DP group was basically intact.When compared with Con group,AKAP150 and PKA expressions were downregulated in rat hippocampal tissues in Pro,DP,and ADP groups,while the expressions of NLRP3,GSDMD,IL-1β,and IL-18 were upregulated(P＜0.05).When compared with Pro group,AKAP150 and PKA expressions were upregulated in rat hippocampal tissues in DP group,while the expressions of NLRP3,GSDMD,IL-1β,and IL-18 were downregulated(P＜0.05).When compared with DP group,AKAP150 and PKA expressions were downregulated in rat hippocampal tissues in ADP group,while the expressions of NLRP3,GSDMD,IL-1β,and IL-18 were upregulated(P＜0.05).Conclusion Dexmedetomidine can alleviate propofol-induced pyroptosis in rat hippocampus at developmental stage,and its mechanism may be related to activating of AKAP150 expression,enhancing of PKA activation,inhibiting pyroptosis-related proteins and the releases of inflammatory factors IL-1β and IL-18.
基金项目
国家自然科学基金(81860234)
贵州省科技计划项目(黔科合基础-ZK[2023]一般226)
维普
万方数据