异氟醚与七氟醚麻醉对新生大鼠小胶质细胞活化和神经干细胞增殖分化的影响及其作用机制  

Effects of Isoflurane and Sevoflurane Anesthesia on Microglia Activation and Neural Stem Cell Proliferation and Differentiation in Neonatal Rats and Its Mechanism of Action

作　　者：赵娟 刘成[1] 贾茜 ZHAO Juan;LIU Cheng;JIA Qian(Department of Anesthesiology,Mianyang People′s Hospital,Mianyang Sichuan 621000,China)

机构地区：[1]绵阳市人民医院麻醉科,四川绵阳621000

出　　处：《联勤军事医学》2024年第7期553-558,共6页Military Medicine of Joint Logistics

基　　金：四川省卫生健康委员会科研课题(19PJ215)。

摘　　要：目的 探讨异氟醚(isoflurane,ISO)和七氟醚(sevoflurane,SEV)吸入麻醉对新生大鼠小胶质细胞活化和神经干细胞(neural stem cell,NSC)增殖分化的影响及其机制。方法 采用随机数字表法将45只6日龄大鼠分为对照组、ISO组和SEV组,每组各15只。所有大鼠均于麻醉开始前腹腔注射5-溴-2′-脱氧尿苷(5-bromo-2′-deoxyuridine,BrdU)50 mg/kg,按照对照组、ISO组和SEV组顺序分别给予30%O_(2)、1.1%ISO+30%O_(2)、2.0%SEV+30%O_(2)吸入,持续吸入4 h,并监测其生命体征和动脉血气。8周龄后,采用Morris水迷宫和T迷宫实验检测大鼠学习和空间工作记忆能力;Western blot检测脑组织小胶质细胞和NSC增殖标志物及血管内皮生长因子受体2(vascular endothelial growth factor 2,VEGFR2)通路相关蛋白表达;免疫荧光法检测大鼠海马区小胶质细胞和NSC增殖分化标志物表达。结果 维持吸入1.1%ISO或2.0%SEV 4 h对新生大鼠生命体征和动脉血气无显著影响。与对照组相比,ISO组和SEV组大鼠成年后的逃逸潜伏期和游泳距离增加、穿越平台次数和交替选择正确率降低(P均<0.01),脑组织中巢蛋白(Nestin)和性别决定区Y框2(sex determining region Y box 2,Sox2)蛋白表达及磷酸化-VEGFR2(phosphorylated-VEGFR2,p-VEGFR2)/VEGFR2比值降低,整合素αM(integrin alpha M,CD11b)、白细胞介素6(interleukin 6,IL-6)和肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)蛋白表达升高(P均<0.05),Nestin^(+)/BrdU^(+)、Sox2^(+)/BrdU^(+)、神经胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)^(+)/Sox2^(+)、双皮质素(doublecortin,DCX)^(+)/BrdU^(+)、洛丝蛋白(Reelim)^(+)/BrdU^(+)和神经元核抗原(neuronal nuclei,NeuN)^(+)/BrdU^(+)细胞占比减少,离子化钙结合适配分子1(lonized calcium-binding adapter molecule 1,IBA1)^(+)细胞占比增多(P均<0.05)。与ISO组比较,SEV组大鼠逃逸潜伏期和游泳距离增加、穿越平台次数减少、交替选择正确率升高(P均<0.05),Nestin和Sox2蛋白Objective To investigate the effects and mechanisms of isoflurane(ISO) and sevoflurane(SEV) inhalation anesthesia on the activation of microglia and proliferation and differentiation of neural stem cell(NSC) in neonatal rats.Methods A total of 45 6-day-old rats were divided into control group,ISO group and SEV group by randomized numerical table method,with 15 rats in each.All the rats were injected intraperitoneally with 50 mg/kg 5-bromo-2′-deoxyuridine(BrdU) prior to the onset of anesthesia,and inhaled 30% O_2,1.1% ISO+30% O_2,2.0% SEV respectively in the order of control group,ISO group and SEV group,continous inhalation for 4 h,and vital signs and arterial blood gas were monitored.After 8 weeks of age,the learning and spatial working memory abilities of the rats were detected by Morris water maze and T-maze experiments;the expression of microglia,NSC proliferation markers and vascular endothelial growth factor 2(VEGFR2) pathway-related proteins in brain of the rats were detected by Western blot;the expressions of proliferation and differentiation markers of microglia and NSC in the hippocampus were detected by immunofluorescence assay.Results Inhalation of 1.1% ISO or 2.0% SEV for 4 h had no significant effect on vital signs and arterial blood gas in the neonatal rats.Compared with control group,the escape latency and swimming distance of adult rats in ISO group and SEV group increased,the number of crossing platforms and the correct rate of alternate selection decreased(all P<0.01),protein expression of Nestin and sex determining region Y box 2(Sox2) and phosphorylated-VEGFR2(p-VEGFR2)/VEGFR2 ratio decreased in brain tissues,the protein expressions of recombinant integrin alpha M(CD11b),interleukin 6(IL-6) and tumor necrosis factor alpha(TNF-α) increased(all P<0.05),the proportion of Nestin^(+)/BrdU^(+),Sox2^(+)/BrdU^(+),glial fibrillary acidic protein(GFAP)^(+)/Sox2^(+),doublecortin(DCX)^(+)/BrdU^(+),Reelim^(+)/BrdU^(+) and neuronal nuclei(NeuN)^(+)/BrdU^(+) cells decreased,the proportion of lonized calc

关 键 词：七氟醚 异氟醚 小胶质细胞 神经干细胞 认知功能障碍 血管内皮生长因子受体2信号通路 

分 类 号：R614[医药卫生—麻醉学]